Animal Adventure Park & Preserve in Harpursville has announced one of the park's giraffes has died said while the giraffe received monthly exams and had been in good health an acute cardiac event is suspected" as the cause of her death A necropsy will be conducted at Cornell University to "determine the specific cause of death," according to the park On Facebook Animal Adventure Park expressed their sadness at Johari's passing More: High winds, snow and freezing rain hit the Binghamton area this weekend: See the totals The average life expectancy for "giraffes in human care" is between 20 and 25 years A giraffe's average life expectancy in the wild is up to 15 years giving birth to calves that reach about 6 feet tall More: Here's everything you need to know about April the giraffe's pregnancy Johari was brought to Animal Adventure Park from another facility in 2019 to be a companion for Tajiri whose 2017 birth was streamed live on YouTube as part of the worldwide coverage of his mother April's first pregnancy at the park Pairing Johari with Tajiri, the park said at the time would help Tajiri start his own family and "fulfill his role in the species management program working towards sustainable and diverse populations." Johari did not give birth to any calves during the years she spent in Harpursville According to their website Animal Adventure is currently home to four giraffes — Oliver Desmond came to Harpursville from a zoo in Milwaukee in 2021, according to the park's Facebook page Tajiri's mother, April, was euthanized in 2021 after suffering from worsening arthritis April had two sons with Oliver — Azizi and Tajiri. Azizi was transported to The East Texas Zoo and Gator Park in 2019 VANCOUVER, BC, Feb. 28, 2025 /CNW/ - Tajiri Resources Corp. (the "Company" or "Tajiri") (TSXV: TAJ) is pleased to announce that in accordance with the rules of the TSX Venture Exchange (the "TSXV") it has received shareholder approval for its previously announced proposed acquisition of the Yono Gold Property located in Guyana The Yono Gold Property is strategically situated just 170 metres from significant gold resources totalling approximately 5.2 million and 2.7 million ounces (Indicated and Inferred) on the adjacent Oko and Oko West Properties owned by G2 Goldfields and G Mining Ventures.1 CEO & President Graham Keevil commented: "As we move forward with the acquisition of the Yono Gold Property I'm thrilled by this strong show of shareholder support we reached a majority of disinterested Tajiri shareholders and secured approval in under 10 days I believe this not only speaks to the enthusiasm investors have with respect to the Yono Project but also the close relationship we maintain with our shareholders We look forward to finalizing the remaining closing steps soon and beginning our work on the property." The Company is working to satisfy the TSXV requirements for the acquisition of the Yono Gold Property under Section 5.7 of TSXV Policy 5.3 the Company is required to: (i) submit a financial plan demonstrating sufficient resources to cover obligations for six months post-closing and the first phase of the recommended work program; (ii) provide required personal information forms for certain insiders as acknowledged by the TSXV with final closing subject to their completion and clearance; (iii) provide a legal title opinion confirming Nebula Resources Inc.'s authority to transfer the Yono Gold Property interest; and (iv) pay the remaining filing fee of $14,700 to the TSXV Until the closing of the acquisition of the Yono Gold Property the Company will provide status updates through follow-up news releases at 30-day and 90-day intervals 1 The Company cautions that information concerning mineralization on adjacent properties is not necessarily indicative of mineralization on the Yono Gold Property or historical estimates from adjacent properties is based on publicly available information which has not been verified by the Company or its qualified person The Company has no interest in or right to explore or develop the adjacent properties referenced in this news release and there is no assurance that similar mineralization or results will be encountered on the Yono Gold Property Investors are urged not to place undue reliance on this information when assessing the Company's prospects The scientific and technical contents of this news release have been reviewed and approved by Dominic O'Sullivan member of the AusIMM and a qualified person as defined by National Instrument 43-101 – Standards of Disclosure for Mineral Projects O'Sullivan is not independent of the Company by virtue of his position as Executive Chairman On Behalf of the Board,Tajiri Resources Corp is a junior gold exploration and development Company with exploration assets located in two of the worlds least explored and highly prolific greenstone belts of Burkina Faso Lead by a team of industry professionals with a combined 100 plus years' experience the Company continues to generate shareholder value through exploration www.tajirigold.com This news release contains "forward-looking information" and "forward-looking statements" (collectively "forward-looking statements") within the meaning of the applicable Canadian securities legislation are forward- looking statements and are based on expectations estimates and projections as at the date of this news release including without limitation; estimated timing geological interpretations relating to the Yono Gold Property and potential mineral recovery processes or results Any statement that involves discussions with respect to predictions future events or performance (often but not always using phrases such as "expects" "believes" or "intends" or variations of such words and phrases or stating that certain actions "might" or "will" be taken to occur or be achieved) are not statements of historical fact and may be forward-looking statements.  Forward-looking statements contained herein are made as of the date of this press release any obligation to update any forward-looking statements whether as a result of new information or if management's estimates or opinions should change There can be no assurance that forward-looking statements will prove to be accurate as actual results and future events could differ materially from those anticipated in such statements the reader is cautioned not to place undue reliance on forward- looking statements Neither the TSXV nor its Regulation Services Provider (as that term is defined in the policies of the TSXV) accepts responsibility for the adequacy and / or accuracy of this release Contact Information: Tajiri Resources Corp., Graham Keevil, President, CEO, 778-229-9602, [email protected] Do not sell or share my personal information: Link IconCopy linkFacebook LogoShare on FacebookXShare on XEmailShare via EmailLink copied to clipboardFour Philly artists receive United States Artists Fellowship awardThis year the award recognizes sculptor Karyn Olivier named four Philadelphia artists as recipients of its prestigious fellowship award granting them each an unrestricted sum of $50,000 The fellows include two Temple University professors — sculptor Karyn Olivier and filmmaker Rea Tajiri; choreographer Nichole Canuso; and veteran muralist and sculptor Cesar Viveros The annual fellowship recognizes 50 artists across the United States and Puerto Rico for “their groundbreaking artistic visions unique perspectives within their fields and evident potential for the award to make a significant impact in their practices and lives,” the organization said in a statement We asked the four new fellows to tell us about the projects they’re developing with this new funding the formerly enslaved woman who saved Stenton House from being burned during the Revolutionary War And one of my favorite pieces was in my own neighborhood—in Germantown’s Vernon Park I made an artwork called The Battle Is Joined The idea was to provoke thought about who decides who (and what) is worth honoring All of these works are built on the shifting sands of history and Philadelphia has so many stories that have been buried beneath the accepted colonial surface Her plans: The fellowship will help me to fund an assistant and hire studio help to manage some of the time-consuming administrative work and research that are integral to an art career It will allow me to continue bringing rigor and enthusiasm to my students who deserve a professor who is focused and not overwhelmed by juggling two full-time jobs How does Philadelphia inspire your artistry Philadelphia is a city steeped in history and rich in cultural diversity and that energy pulses through every neighborhood I’ve had the privilege to work in What stands out most is the city’s true embrace of the term “Sanctuary City.” People here offer a welcome His plans: I am currently working on a series of projects that center on the immigrant experience and the cultural richness we bring to Philadelphia is an immersive art installation that serves as a tapestry of immigrants transforming the everyday into something extraordinary I plan to bring more of this work into public spaces inviting larger audiences to reflect on the surrealism in our lives I’ll continue introducing new projects like La Cruceta Spanish and Nahuatl languages while engaging the broader community in meaningful conversations Neighborhood: Between Old City and Northern Liberties Philadelphia has such a rich physical landscape we’re on the territory of the Lenapehoking then there are the confluences of several creeks and rivers of the Wissahickon Cobbs Creek which gives it a strong energy The art culture here is both international and hyperlocal and intimate; it’s very scrappy and resourceful I’m very appreciative of the accessibility to have in-depth conversations with other artists here I also appreciate the community media spaces like Blackstar Lightbox – spaces where we can convene and engage Her plans: I’m putting the funds towards a current project It’s based off of a rediscovered photography book composed of my father Vince Tajiri’s personal work he was documenting the resettlement of Japanese Americans into Chicago after the WWII U.S He submitted the book for publication but it was rejected and interpreting his life and experiences based off of an archive of my father’s writing just discovered in the last year I was born and raised here; it’s home base for my heart and my career The trajectory of my practice has been shaped by the collaborators audiences and residents I’ve encountered here I seek to soften the boundary between audiences and performers to invite tender exchanges and moments of reflection allowing me to learn from and respond to buildings and their people iterative process is continually expanding my own perspective on my environment Philadelphia’s way of carrying its history informs how I engage with sites around the world Her plans: I’m currently embarking on an immersive performance installation called Lunar Retreat named after the distance growing between the Earth and the Moon It will be an interactive space that invites rest and reflection offering participants invitations to process loss and transformation These interviews have been edited and condensed for clarity The Yono Property is strategically situated just 170 metres from significant gold resources totalling approximately 5.2 million and 2.7 million ounces (Indicated and Inferred) on the adjacent Oko and Oko West Properties owned by G2 Goldfields and G Mining Ventures Barry & Associates Limited in connection with the Company's proposed acquisition of a 65% working interest in the Yono Property It provides a comprehensive analysis of the Yono Property confirming key geological features previously disclosed by the Company The full report is available on SEDAR+ at www.sedarplus.ca "The Yono Property is strategically located very close to adjoining it could be very challenging and restrictive to mine these deposits without having access to the Yono Property." 1 Other noteworthy excerpts from the report include: "The Yono Property has excellent potential to host mineralization similar to that on the adjoining properties and similar to other known gold deposits in the region The Yono Project is a Property of Merit and an extensive multi-phased exploration program is warranted.2 The Company notes several gold targets on the Yono Property have been identified in the report Of prominence is the potential for strike extensions of the western portions of the higher grade (6.38-9.3g/t Au)3 Oko Main Zone to extend into the eastern portion of the Yono Property "The most recent published drill results by G2 Goldfields (G2 Goldfields Inc 2025) appears to show that the Ghanie Zone and OMZ Shear 1 are contiguous and potentially Shear 1 and Shear 3 coalesce between Ghanie and OMZ In this interpretation Shears 4 & 5 are open to the south of OMZ and substantively untested and potentially project into the Yono Property along its eastern boundary somewhere between its northeastern and southeastern corners."4 "Favourable sedimentary and volcanic rocks that host gold mineralization on adjacent properties have been mapped in the centre of the Yono Property."5 Figure 1 reproduced from Figure 21 of the report illustrates these relations CEO & President Graham Keevil commented "We're extremely pleased to have completed this most crucial step in the Yono Property acquisition approval process the independent technical report further serves to confirm our belief that the Yono Property could be highly prospective and a potentially invaluable part of the developments at the two neighbouring gold projects." _____________________________1 Page 97- (last paragraph) of the NI 43-101 Technical Report on the Yono Property 2 Page 2- (Section 1.6) of the NI 43-101 Technical Report on the Yono Property 3 The potential quantity and grade of mineralization disclosed in this news release are conceptual in nature There has been insufficient exploration to define a mineral resource and it is uncertain whether further exploration will result in the target being delineated as a mineral resource The potential quantity and grade disclosed in this news release are based on a combination of factors and comparisons to known mineralized zones in the region These factors indicate the presence of mineralization; however is required to determine whether a mineral resource can be defined Investors are cautioned that the exploration target is not being reported as part of any current mineral resource estimate and should not be relied upon as an indication of future resource potential 4 Page 52 (last paragraph) of the NI 43-101 Technical Report on the Yono Property 5 Page 8 (last paragraph) of the NI 43-101 Technical Report on the Yono Property The technical contents of this news release have been reviewed and approved by Dominic O'Sullivan www.tajirigold.com including without limitation; anticipated results of geophysical surveys or drilling programs geological interpretations and potential mineral recovery processes Contact Information: Tajiri Resources Corp., Graham Keevil, President, CEO, 778-229-9602, [email protected] NoDQ.com: WWE and AEW Coverage watch Tajiri challenge Triple H for the World Heavyweight Championship at a January 2003 WWE Live Event in Japan Find the latest Superstar gear at WWEShop: http://shop.wwe.com VANCOUVER, BC, Dec. 4, 2024 /CNW/ - Tajiri Resources Corp. (the "Company") (TSXV: TAJ) is pleased to announce that it has received final acceptance from the TSX Venture Exchange ("TSXV") and has closed its non-brokered private placement offering of units (the "Offering") previously announced September 23 The Offering consisted of two tranches (see news released October 09th and November 06th 2024) totalling 19,894,000 Units priced at $0.05 per Unit for aggregate gross proceeds of $994,700 Each Unit consisted of one common share (each a "Common Share") and one common share purchase warrant (each Each warrant is exercisable by the holder to acquire one Common Share at a price of $0.10 for three years from the date of issuance subject to the Acceleration Right (as defined herein) the closing price of the Company's common shares on the TSXV for any ten (10) consecutive trading days equals or exceeds $0.25 upon providing written notice to the holders of Warrants to accelerate the expiry date of the Warrants to the date that is thirty (30) days following the date of such notice which may provided by way of a news release (the "Acceleration Right") the Company paid a total of $49,679 cash and issued 992,580 non-transferable Broker Warrants bearing the same terms as those attached to the units to certain finders in consideration for introducing certain purchasers to the Company Pursuant to applicable Canadian securities laws all securities issued in connection with the Offering are subject to a four (4) month hold period from the date of issuance Proceeds from the Offering for the exploration and development of the Company's mineral properties and for general working capital purposes with roughly $30,000 of the proceeds raised in the first tranche to be used toward the acquisition of the Yono Gold Property in Guyana The Company will not be proceeding with the previously announced third tranche of the placement at this time and the placement has now officially been closed is a junior gold exploration and development company with exploration assets located in two of the worlds least explored and highly prospective greenstone belts of Burkina Faso  Led by a team of industry professionals with a combined 100 plus years' experience the Company continues to generate shareholder value through exploration including but not limited to expected size of the second tranche the proposed use of proceeds of the Offering the anticipated closing date of the second tranche of the Offering the closing Project acquisition and receipt of the approvals required and related thereto are forward-looking statements and are based on expectations "might" or "will" be taken to occur or be achieved) are not statements of historical fact and may be forward-looking statements Forward-looking statements involve known and unknown risks uncertainties and other factors which may cause the actual results performance or achievements of the Company to be materially different from any future results performance or achievements expressed or implied by the forward-looking statements. Forward-looking statements contained herein are made as of the date of this news release and the Company disclaims  There can be no assurance that forward-looking statements will prove to be accurate the reader is cautioned not to place undue reliance on forward-looking statements Neither the TSX Venture Exchange nor its Regulation Services Provider (as that term is defined in the policies of the TSX Venture Exchange) accepts responsibility for the adequacy or accuracy of this release Contact Information: Tajiri Resources Corp., Graham Keevil, President, CEO, 778.229.9602, [email protected] A never before seen match featuring Triple H in Japan has been released from the WWE Vault featuring an entrance with Stephanie McMahon While he may currently appear on television as a WWE authority figure Paul ‘Triple H’ Levesque had a storied in-ring career before fully focusing on being an executive One such match has now been released from the WWE Vault on YouTube featuring Triple H wrestling in Japan Not only the match is featured in the clip with Stephanie McMahon making an introduction as well as ring announcing from Ric Flair The ‘never-before-seen footage’ features Tajiri challenging Triple H for the World Heavyweight Championship at a January 2003 WWE Live Event in Japan Spoiler alert – the match ends with The Game reversing an attempted finisher from Tajiri into a Pedigree to retain the WWE World Heavyweight Championship You can see the entire match in the video below You can keep up with all the latest news from around WWE at this link. To make sure you stay up to date with all the biggest wrestling and WrestleTalk news, follow us on BlueSky by clicking this link © Trident Digital Media Limited. All rights reserved. Read our Privacy and Data Policy Designed and Developed by Arren Marketing Tajiri is still going strong in the Japanese wrestling scene competing on a very regular basis throughout 2024 Surprisingly his last WWE appearance was not that long ago either competing in the Cruiserweight Classic back in 2016 as random as it would first seem Tajiri is arguably one of the stars most ready to go for a WWE Royal Rumble cameo Bringing the former Cruiserweight and Tag Team Champion back to WWE would be a joy for everyone to see Please view the main text area of the page by skipping the main menu. The page may not be displayed properly if the JavaScript is deactivated on your browser Japanese version We use cookies to personalize content and ads and to analyze our traffic and improve our service In the opening scene of the first film I saw by Rea Tajiri a match is lit and raised to a piece of gray cloth in the dark my grandfather burned the family’s possessions in the strawberry fields behind the house.” The film was Strawberry Fields (1997) It was the first film I saw with an entirely Japanese American cast and it was the first film I saw about JA incarceration that centered the experience and perspective of a descendant of the concentration camps the past is extinguished in a desperate yet clear-sighted act and in that instant the primary condition of the family in Strawberry Fields is one of loss was incarcerated in the Poston (Arizona) camp—what follows is a pilgrimage to the source of that loss is a documentary portrait of Rose and her descent into dementia but it feels limiting to say that because all of Rose’s previous selves “Rose was layering people from the past with the present,” Tajiri says in the film “It was one of her ways of time-traveling and making it possible for all of us to meet.” This is a beautiful expression of what it means to undergo the long transformation through dementia into the afterlife It is also a perfect definition of the films of Rea Tajiri Brandon Shimoda I have seen Wisdom Gone Wild several times and what struck me this time is the voice at the beginning: a woman calling out the name Akiko—intimately each time more insistently: Akiko … Akiko … Akiko Its invocation is profoundly intimate yet haunting to begin with what I am assuming is your grandmother’s voice Rea Tajiri The film is about aging and who we are at different stages and moments across the span of a lifetime I wanted to weave this specific element into the film that would show aspects of a parent’s life and how a child might remember them across a lifetime There was a particular moment when I was six and heard my grandmother call my mom by her Japanese name We were traveling for a family reunion from Chicago—city of resettlement—to Fresno In that split second I sensed my mother as a child of my grandmother and felt my grandmother’s mothering energy It was startling because it was an entry point into my heritage hearing this name and my grandmother’s utterance My family had no heirlooms; anything passed along was lost during incarceration So that utterance came back to me when I was looking through archival photos of my mother’s childhood I realized I could embody and perform my grandmother for the sake of the film you covered the beauty shop in Rose’s assisted living facility with plants It feels like she is transitioning through the final realization of herself while returning to something essential Where did the idea come from to create this dreamlike space I had a dream about a group of Nisei elder women gathered in a backyard around beauty shop stations led by a beautician ringleader who was orchestrating them The dream sprung from Rose; she was a beautician and loved gardening pre-dementia There were many moments off camera when Rose would do or perform things; but I couldn’t just say “Do that again!” I thought that if I created a specific set piece it could spark her imagination to run freely who were doing production design for big-budget Hollywood films—including Pulp Fiction for which they won an Academy Award for Best Production Design I remember once when I came into Rose’s room the light was filtering in from the window through a tree outside and she started reciting a poem off the top of her head She would have been if she had opportunities and support BS These layerings and ways of time-traveling—you inviting your mother into your dream maybe your mother inviting you into her dreams too—make me wonder: What do you think your mother’s dementia revealed or permitted of her inner life that might have remained inaccessible Did you experience aspects of her dementia while making Wisdom Gone Wild that you did not otherwise see RT When someone you’ve known your whole life begins to transform through dementia Sometimes it feels like their neuroses and their fears are amplified too The person loses abilities that they never gave a second thought to—my mother completely forgot about gardening She’d look at plants and flowers with complete disinterest What if I accept the reality she’s creating it was surprising what worlds she would build And sometimes little things would slip out A name of a place I never heard her mention names of elementary school teachers and classmates or it allowed me to see what she didn’t like growing up I don’t want to seem like I am romanticizing dementia but I do want to model another way of engaging with an elder who has dementia Aren’t we all trying to get back to our authentic selves Brandon Shimoda is the author of several books most recently The Grave on the Wall (City Lights) Click here to read our spring 2025 issue, featuring Caught by the Tides' Jia Zhangke and Zhao Tao, our annual spotlight on locations and more... Rea Tajiri’s Wisdom Gone Wild takes a hard look at a difficult subject Tajiri’s 93-year-old mom Rose is a witness to the US’s dark concentration camp history having been incarcerated along with the rest of her Nikkei farming family during the Second World War Tajiri goes (and takes us) on a decade-plus nonlinear cinematic journey— neatly paralleling Rose’s own thought process as the veteran filmmaker’s mom began her dementia decline at the age of 76—or should I say dementia “reinvention.” For far from being a tragic story about “losing” one’s mind Wisdom Gone Wild is actually a celebration of life in all its remarkable phases as both Tajiri and her mother have decided to embrace the new woman Rose is forever transforming herself into an identity complete with different surname and metaphorical past Herzog’s “ecstatic truth” in perpetual motion Just prior to the doc’s November 20th airing on POV Filmmaker reached out to the multi-award-winning director (History and Memory Strawberry Fields) and interdisciplinary artist whose choice to put her career on hold to care for her main character seems to have paid off in spades Filmmaker: Had you always planned on making a feature-length doc about your mother—and specifically her WWII incarceration how did your vision for the film change as her dementia progressed I was about to travel to the Venice Film Festival for my film Strawberry Fields I was devastated because we spoke excitedly about this one day then the next day we were to follow up and she had completely forgotten (to the point where she thought I was someone else impersonating her daughter) everyone around me encouraged me to film the experience but this idea seemed unethical and a violation of my mother’s privacy I traveled sometimes once a month from NYC to Los Angeles I leaned heavily into my Buddhist practices I sat with the loss and contemplated what it was that I was losing and you have to let go and accept loss as a part of life Once I began that journey something shifted I discovered ways of connecting with my mother that were quite profound which she enjoyed—not in an exhibitionist way but it was like we were photographer and model We sent these to family and friends to share the process Later I started filming little clips with a point-and-shoot I ended up with a personal archive of fragments; it was not intended to be a full film back then I had learned a lot by then and felt like I could share another perspective on living with dementia that others would find useful one that was not part of the public dialogue around dementia and caregiving I hired a DP for two days using money from a tax refund Our first shoot was at the Los Angeles County Museum of Art and that shoot went so well I knew this would be possible I felt an urgency to make this film before it became too late “By any means possible film her and keep going even if you make this on an iPhone.” That was inspiring Filmmaker: I believe you actually put your career on hold to care for your mother—an experience that seems both common and yet rarely depicted onscreen (though I’m not sure why) So was filming her a way to continue your craft while caregiving—or were you focused on preserving personal memories Tajiri: I worked as an industrials producer which paid well and allowed me chunks of time off so that I could travel to Los Angeles from New York to care for her When you’re stepping away from a career you’ve staked your identity on But in that liminal space I really began to see my life as not just about a career and caring for another person took on great importance I learned some profound lessons about surrender and acceptance Filmmaker: Do you feel that being an experimental artist comfortable with nonlinear narratives allowed for a greater openness to your mom’s “reinvention”—or was acceptance still a hurdle you had to overcome Tajiri: I actually thought that I could make a conventional I thought at first that verisimilitude and “realism” would be necessary for this story But the nature of the material was too fragmentary and I didn’t like the results I was also trying to avoid certain narrative cliches about dementia—the tragedy (“she was once this amazing person but look at her now”) or the mother-daughter redemption story Those types of narratives are so constrained and couldn’t contain all the things I wanted to touch on I decided to frame this around “time travel,” in order to mirror my mother’s logic—which was always jumping in and out of different eras—and how that gave me insight into her life She shared stories about her past that I’d never heard before they were fascinating in how they intersected with key historical events and were meaningful her reinvention was a key part of this story and a huge inspiration to me Somewhere she found the “creativity” to make up a new identity that She would tell everyone that my father never existed that I was not her daughter but her sister and she was a professor of art history and on and on I had become a professor of film in the years I took care of her I saw how her new identity contained seeds of her desires and dreams Filmmaker: I remember asking Maite Alberdi (back during our Sundance interview for The Eternal Memory) how she navigated the issue of consent when filming with a character who may not always be able to grant it How did you go about ensuring your mom was always okay with having such intimate moments in her life revealed onscreen and I thought about this carefully throughout the filming and in the edit room What I decided was: I lived this experience with my mother intimately for 16 years and people living with dementia are not well represented in films but I also didn’t want to take advantage of a situation Dementia is a complicated disease; it needs to be seen more and nuanced portrayals are necessary my mother was capable of objecting to things she did not want to do Some might argue she wasn’t capable of understanding If I gave consent to be filmed by someone and signed a piece of paper does that make it okay if a month later I change my mind We need to have formal protocols in place to protect people but we also need to be careful about each situation (One festival rejected the film on the basis that they felt I created an exploitative representation of my mother and that she could not have been capable of giving her consent.) I knew my mother well enough to know what she would object to I think my mother wanted me to succeed as a filmmaker That meant sharing a meaningful and important story to the world told from this unique perspective She and I constantly discussed filming and what I would be doing with the footage On some deep level my mother trusted me; she knew my films before she developed dementia There is one scene in which she asks me to stop filming her and I don’t I left that in the film to show what happened when I didn’t listen to her We don’t get to see elderly people getting angry I feel that a film about caregiving and dementia by default has to look honestly at the intimacies and intricacies of the process My father was a photographer and photographed us throughout our lives In the end we saw it as a part of his way of relating to us creating art in the moment from the life he was observing and even became performative for the camera I personally don’t like being filmed by other people that I don’t know Filmmaker: What lessons did you (and your family) learn that you hope audiences likewise will take away from the film Tajiri: Caregiving can be a profoundly life-changing experience and they can provide deep connection and deeper insight It is frustrating—it will foreground whatever tensions or difficulties exist in your relationship it is a lesson in setting aside one’s ego for another person It may not always be the right choice for some people maybe this will be too painful and impossible; if you are too triggered I saw it as taking the time to acknowledge life itself I had to deeply consider another person’s wellbeing [and] the opportunity to make someone else’s life comfortable and meaningful There comes a time in every wrestler's career when they must take a long, hard look in the mirror and realize it may soon be time to hang up the boots. For TAJIRI, whose several-year run with All Japan Pro Wrestling as both wrestler and booker just came to an end At an introductory press conference for his arrival to Fukuoka-based promotion TAJIRI remarked that his run with Kyushu Pro would and I look forward to working with you," he said and I don't think I have much time left in my wrestling career so I think that joining Kyushu Pro Wrestling is the beginning of the end TAJIRI currently holds the Kyushu Pro Championship which he won from Kodai Nozaki on January 3 in his debut for the promotion after conversing with Kyushu's owner and promoter Ryota Chikuzen the opportunity to hold titles and help younger talent was instrumental in convincing the 52-year-old to come aboard I want to be the champion and have hot matches I want to nurture the younger generation," he added he said that he would be very interested in having me work in Kyushu and I thought that this would be an organization where I could demonstrate my abilities to the best of my ability." It should be noted that the former ECW, WWE, and MLW veteran didn't give a precise timetable for the final wind-down of his career or if any sort of retirement tour might be in the works « Back It’s not often a birthday party for a one-year-old will be seen around the world But not every tot is the offspring of April the giraffe the social media sensation who gave birth to son Tajiri as millions of viewers watched livestreamed pregnancy and labor catapulted Animal Adventure Park in Harpursville to unlikely fame The idea to livestream April’s pregnancy and birth came in response to local interest in the giraffe’s pregnancy Animal Adventure Park posted an update about April’s pregnancy on Facebook The post not only satisfied the curiosity of local giraffe fans but was seen by 317,000 people worldwide a single day “No one here expected it,” said Patch “But the fandom grew internationally.” As April’s belly expanded, so did her following. Updates were posted frequently on Facebook and Twitter, and in February 2017, the Giraffe Cam went live on YouTube More: April the Giraffe rules Animal Adventure (and YouTube) in 2017 More: April has been cleared for pregnancy, but some people aren't happy More: After surgery, great expectations for Animal Adventure park owner's daughter it was in front of millions of YouTube and Facebook Live viewers watching from as far away Asia and Australia Named Tajiri – which means “hope” in Swahili – the new baby giraffe stood a gangly 5’9” and was instantly famous A year later, Tajiri stands 10 feet tall and weighs around 1,000 pounds “It’s an expedited growth rate,” said Patch noting that mature giraffes average about 18 feet in height Patch also said the Tajiri is becoming more independent of his mom April as he weens The tall tot also shows personality characteristics of both his mother and father “Like his father Oliver, Tajiri is outgoing The entire Harpursville giraffe family continues captivate fans worldwide An international influx of giraffe fans lined up along locals last season to feed the giraffes carrot treats and the Giraffe Cam still attracts thousands of viewers each day With Animal Adventure Park doesn’t open until May 16, Tajiri will be celebrating his birthday on Sunday with a party in the giraffe barn for the giraffe family “Like any one-year-old, Tajiri will get cake,” said Patch of the party Tajiri's cake will be made specially formulated giraffe food and  topped with a carrot-and-cauliflower icing And like his mom April’s pregnancy and his birth, Tajiri's enjoyment of the cake will be streamed live on the Giraffe Cam Your request appears similar to malicious requests sent by robots Please make sure JavaScript is enabled and then try loading this page again. If you continue to be blocked, please send an email to secruxurity@sizetedistrict.cVmwom with: Rea Tajiri is an award-winning interdisciplinary artist and educator who creates installation non-traditional storytelling forms to encourage dialog and reflection around buried histories Tajiri is a Sansei who grew up in Rogers Park She earned her BFA and MFA degree from the California Institute of the Arts where she studied post-studio art two early shorts were included in the Whitney Biennials of 1989 and 1991 History and Memory: For Akiko & Takashige went on to receive the Distinguished Achievement Award from the International Documentary Association and a Special Jury Award from the San Francisco International Film Festival and Best Experimental Video from the Atlanta Film and Video Festival This documentary short has screened in over 250 venues around the world History and Memory is distributed by Women Make Movies You can also watch it on the Criterion Channel In 2014 Rea Tajiri completed her feature-length documentary Lordville This hybrid documentary was nominated for a Grand Jury Prize at CAAMFest The film screened recently at Yerba Buena Arts Center Wisdom Gone Wild won the Audience Award and Honorable Mention Jury Award for Best Documentary at the 2022 Blackstar Film Festival.  and why did you choose a career in documentary filmmaking?  Rea Tajiri: I consider myself a visual artist – that’s where I started from I went to art school at Cal Arts in the late 1970s But entering documentary filmmaking was an organic evolution so I grew up surrounded by outtakes and miscellaneous random photos from whatever he was processing in his basement darkroom– I was constantly playing with these images that had this sense of the uncanny–without context as to who was in them or where they were taken I think this heavily influenced my work because I draw from photography and work from the archive a lot The evolution from video artist to documentary filmmaker happened because I wanted to come to terms with Japanese American incarceration- what this meant for my family needing to build the context - piecing together stories and clues like the detective This subject necessitated a documentary format IDA: Tell us a little about your recent film Wisdom Gone Wild RT: My film is centered around my mother who was diagnosed with dementia in 1999 It’s a memoir that is immersive and experiential I had to shoot most of it on the fly as I was taking care of her in hospice and later I created a set-piece beauty shop tableau for her I made that space for her based on her life as a beautician and her love of gardening Then I brought her in to see how she would respond and to our surprise and delight – I utilized family archives to bring the viewer into her past– but I had a delicate balancing act to perform: I wanted to show the state of mind - elder consciousness - the free association and the ‘time traveling consciousness’ that elders engage in and I didn’t want to devolve into a kind of sentimental family narrative voice recordings and photos and observational footage was very intricate I want to talk about the things we don’t see often in films about aging that care can be very active if you use imagination and the sensory - follow the mind of the elder Blackstar was the perfect place to open this film The connection and the space of intimacy the film creates – the sharing of histories across generations and ancestral knowledge is something that resonated with this audience and we won two awards Mostly getting this longitudinal project off my shoulders and into the world I shared different edits with students which is something I have never done but I wanted to show them how you have to hang in there with a project What is important is that the film connected to people in a very deep way - many people told me they see their relationships to the elders in their lives differently Some people who were caregivers to their parents said they’d never seen a film that shows that very intimate private space that unfolds when you take care of someone day to day - they were grateful for that I think knowing your film has communicated something beyond what you were trying to do gives one a sense of completion I’m hoping for new opportunities and support I have a new body of work for documentary installation that I’m eager to begin IDA: What piece of advice would you give to emerging filmmakers that are from historically excluded backgrounds and who are new to the documentary field?  RT: This is a good time for BIPOC / Filmmakers – there are so many more resources and labs and support communities built up than when I was starting out I guess it’s still terrifying if you don’t feel like you fit If you can find a mentor or a small group of people that you trust and respect your way of storytelling It's important to understand the history of documentary filmmaking and the movements and to watch as much as you can even if it's ‘slow’ keep taking photos and make time to watch your footage and share work with people who you trust and can share equipment and space with Don’t be afraid to ‘break the language’ a bit If your stories don’t necessarily fit into some of the existing storytelling forms It’s good to experiment but also to gain better clarity around why you’re doing what you’re doing IDA: What are some of the changes that are happening in the field that you are excited about RT: Everywhere I hear from friends across the country that cultural institutions are in chaos – We are in the process of dismantling years of white supremacy white leadership - and there’s a transition into different ways of leadership and running an organization We’re going to have to move through this to get to another more equitable "Wisdom Gone Wild will be playing this November at festivals -- Check out the website for festival screening dates and times." Would you like to receive event invitations and other updates from the International Documentary Association virtual on Zoom or in-person at IDA Office in Los Angeles © 2024 International Documentary Association Privacy Policy The Bourse de Commerce will feature an evening devoted to the new contemporary electronic scenes ranging from the radical sounds of singeli from Tanzania to the fast-paced trans bail funk from Brazil and to futurist punk from Barcelona The evening opens with Franco-Guadeloupian producer Mookie a member of the cutting edge Jokkoo Collective in Barcelona a major figure in São Paulo’s underground scene and the mother-daughter singeli duo from Dar es Salaam The evening will conclude with a set from DJ Travella A bar will be open during the performances Mookie is a DJ and Franco-Guadeloupian producer A key figure in Barcelona’s Afro-diasporan experimental collective Jokkoo where his love for punk music melded with electronica where he forms part of various musical and visual arts projects Brazilian MC and DJ Badsista is one of the major figures of the underground musical scene in São Paulo Known for playing sped-up Brazilian trance and funk Badsista performs high-energy sets featuring rave Committed to fighting for inclusiveness in the music scene she is the founder of the LGBTQIA+ collective Bandida She is also a member of the collective Tormenta The mother-daughter duo of Queen Asher & Rehema Tajiri – respectively MC and DJ-producer – major figures in the singeli scene of Dar Es Salaam will give their first-ever live performance in France Rehema Tajiri’s performing career began in the 2000s as a singer and dancer in the Congolese and Tanzanian rumba scene She ventured  into zouk and then singeli Tanzania that combines pop music with traditional polyrhythms an ode to dancing for its ultra-fast rhythms Tajiri was invited by the Ugandan festival Nyege Nyege where singeli was introduced to the the whole world by Boiler Room Queen Asher is the first female singeli DJ they form a fantastic duo that has pushed singeli to an entirely new level The duo debuted at the Nyege Nyege Festival in 2022 and the CTM Festival in January 2023 Their first album is slated for release at the end of this year The event was conceived together with French musician Low Jack.  Open Monday to Sunday from 11:00 am to 7:00 pmLate opening on Friday until 9:00 pmClosed on Tuesdays and May 1Late opening every first Saturday of the month from 5pm to 9pm Practical information Get the latest news from the Bourse de Commerce this article started life as an article on Satoshi Tajiri upon doing research to focus on developers with autism I noticed something: autism is not really discussed in the games industry some developers will actively avoid mentioning the subject due to how many gamers will still use autistic as an insult I thought that I would try to help break this stigma a bit there’s no shame in any form of neurodivergence it’s not a disease when you can’t help the fact that your brain is simply wired that way let’s begin with the original focus of my article Pokémon has been a series that has defined many childhoods and I recall how it was one way for me to bond with not only family but also friends one commonly held belief was that Satoshi Tajiri was autistic I discovered that he was simply commonly believed to have been on the spectrum due to his bug-collecting hobby this was a blow to the original highlight of my article; however I thought I would do research into any other developers with autism with some even saying that Hidetaka Miyazaki (of FromSoftware) has autism so I’m also going to consider this untrue unless claimed otherwise I found there are few developers who are open about such things as neurodivergence you’ve most likely heard of being called autistic as an insult all I realised while I was researching this article was that despite gaming being a great outlet for kids and adults with autism it’s not always the friendliest space out there It doesn’t help that many people assume that being autistic means that you are inherently good with technology and maths and it leads to the idea of the “wunderkind” which is an equally damaging stereotype for those with autism As someone with a family history (and a pending diagnosis) on the spectrum it’s more damaging to claim that kids with autism are essentially geniuses in the field they may have a hyper fixation on While this began as an article to celebrate autism in gaming I’ve instead chosen to focus on raising awareness and keeping an open mind about the subject I’ve come to realise that the one way to break the stigma of the subject is by being open that misconceptions are more damaging than outright hostility let’s take a moment to consider how we can be more open about autism in gaming and make an environment that’s welcoming of everyone (and if the internet can stop using autistic as an insult Writing about all sorts like a liquorice allsort The Japanese Buzzsaw had his fair share of WWE thrillers.. Off the back of an impressive performance in the Cruiserweight Classic tournament WWE have apparently offered Tajiri a contract to stay on with the company Whether that's as a trainer or an active in-ring performer remains to be seen but it does mean more Tajiri and that's no bad thing really like Tajiri and would go as far as to say he's one of my favourite wrestlers his comical facial expressions and his high-pitched shrieking noises I loved him wether he was tearing it up with his fellow cruiserweights on Smackdown or teaming up with William Regal on Raw The Japanese Buzzsaw hasn't been seen in a WWE ring in over a decade he made a one-off appearance at the second ECW One Night Stand pay-per-view in June 2006 but wrestling all over the world and even forming his own promotion Smash in Japan (which ran from Feb 2010 - March 2012) I've identified what I consider to be eight brilliant matches for your viewing pleasure these eight bouts should give you an idea of the calibre of performer that Tajiri is and why you should be excited for his imminent return to screens Before making his way to Mexico and then ECW Yoshihiro Tajiri was a young boy for several promotions before he found himself working for a small Japanese independent promotion called Big Japan Pro Wrestling who were famed for their so-called 'death matches' Tajiri impressed the mighty New Japan Pro Wrestling enough to give him a chance but a match against one of their most established Junior Heavyweights The young-looking Tajiri managed to p*ss off the grumpy Otani before the two had even locked up by offering his hand and then rescinding it as Otani went to shake it The story of the match was very much the young buck (no not those ones) being taken lightly by the more experienced veteran only to surprise him with his immeasurable fighting spirit and total lack of fear He was totally trying to make a name for himself here and started the match by dominating with kicks and submission holds Otani regained his composure and trapped Tajiri in an ankle lock but the plucky youngster came back with some slaps and a palm strike followed by a lovely bridging dragon suplex and an asai moonsault drawing a reaction from the Tokyo Dome crowd Otani inevitably cuts him off and lands a powerbomb allowing Tajiri one more moment of hope before finishing him off for good with a springboard spinning wheel kick but he made a lasting impression on the fans and NJPW officials who brought him back in the spring for the Best of the Super Juniors tournament WhatCulture is part of Future plc, an international media group and leading digital publisher. Visit our corporate site (opens in new tab) ©Future Publishing Limited Quay House England and Wales company registration number 2008885 Metrics details cAMP is a universal second messenger regulated by various upstream pathways including Ca2+ and G-protein-coupled receptors (GPCRs) a sensitive genetically encoded green cAMP indicator that outperformed its predecessors in both dynamic range and cAMP affinity Two-photon cAMPinG1 imaging detected cAMP transients in the somata and dendritic spines of neurons in the mouse visual cortex on the order of tens of seconds multicolor imaging with a sensitive red Ca2+ indicator RCaMP3 allowed simultaneous measurement of population patterns in Ca2+ and cAMP in hundreds of neurons We found Ca2+-related cAMP responses that represented specific information such as direction selectivity in vision and locomotion our multicolor suite will facilitate analysis of the interaction between the Ca2+ GPCR and cAMP signaling at single-cell resolution both in vitro and in vivo cAMP is regulated by these multiple upstream signaling pathways and modulates diverse cellular functions through cAMP-dependent kinases technologies to visualize the spatiotemporal dynamics of cAMP in vivo are crucial for biological research in various organs or species a green cAMP indicator with a high dynamic range and more than 4-fold higher cAMP affinity than the existing green cAMP indicators Dual-color imaging of RCaMP3 and cAMPinG1 revealed dynamic interaction and information flow between Ca2+ we demonstrated the application of cAMPinG1 imaging in cultured cells for studying GPCR biology a, Top, primary structure of cAMPinG1. Bottom, tertiary structures of cAMP-binding PKA-R1α (PDB 1RGS) with cAMP and cpGFP (PDB 3WLD calmodulin and RS20 domains are hidden) are shown with the linkers between the two domains (dotted lines) In vitro screening results of 251 variants with resultant cAMPinG1 Excitation and emission spectra of cAMPinG1 in cAMP-free and cAMP-saturated states ΔF/F of cAMP sensors for cAMP change from 0 to 300 μM in HEK cell lysate at 490 nm (left) or 450 nm (right) of excitation Tukey’s post hoc test following one-way ANOVA gCarvi versus cAMPinG1 at 490 nm: P = 1.5 × 10−14; G-Flamp1 versus cAMPinG1 at 490 nm: P = 2.4 × 10−8; cAMPinG1 versus cAMPinG1mut at 490 nm: P = 1.9 × 10−15; gCarvi versus G-Flamp1 at 450 nm: P = 1.7 × 10−9; gCarvi versus cAMPinG1 at 450 nm: P = 3.7 × 10−6; G-Flamp1 versus cAMPinG1 at 450 nm: P = 1.9 × 10−7 gCarvi versus cAMPinG1: P = 1.5 × 10−14; G-Flamp1 versus cAMPinG1: P = 3.1 × 10−10 488-nm and 405-nm excitation lights were used in turns for ratiometric imaging in HEK293T cells Representative images of HEK293T cells expressing cAMPinG1 excited by 488 nm (left) and 405 nm (right) before (top) and after (bottom) 50 μM forskolin (FSK) application ΔF/F of cAMPinG1 (left) and the inactive mutant cAMPinG1mut (right) in response to 50 μM forskolin application The ratio (488 ex/405 ex) was calculated with fluorescence excited by 488 nm and 405 nm All shaded areas and error bars denote the s.e.m Source data Schematic of the experimental procedure of cAMPinG1 somatic imaging cAMPinG1 or cAMPinG1mut was delivered to neurons in L2/3 of the mouse V1 by in utero electroporation A representative in vivo two-photon fluorescence image of cAMPinG1 Single-trial cAMP traces of 3 representative cells The orange box indicates the timing of the stimulus Averaged traces of somatic signals of cAMPinG1 (left) cAMPinG1mut (left) and G-Flamp1 (right) in response to airpuff stimulation Averaged ΔF/F of cAMPinG1 and cAMPinG1mut in response to airpuff stimulation cAMPinG1 versus cAMPinG1mut: P = 6.9 × 10−10 Half-decay time of somatic cAMP transients in response to airpuff A representative image of cAMPinG1 imaging in spines and their shaft Representative traces of a dendritic shaft and two spines The orange square indicates the timing of the stimulus Averaged ΔF/F of cAMPinG1 in dendritic shafts and spines Boxes indicate the 25th and 75th percentiles and whiskers indicate the total range of data Source data Then, we imaged cAMPinG1-NE in dendritic spines and shafts in vivo under lightly anesthetized conditions (Fig. 2g). We observed sensory-evoked cAMP transients in both dendritic spines and shafts (Fig. 2h,i) which requires advanced sensitivity of the indicators such as spine imaging a, Top, primary structure of RCaMP3 with amino acids mutated relative to R-GECO1 (in R-GECO1 numbering). Bottom, tertiary structures of R-GECO1 (PDB 4I2Y) with amino acids mutated in RCaMP3 (sphere) jRGECO1a versus RCaMP3: P = 5.8 × 10−11; XCaMP-R versus RCaMP3: P = 4.0 × 10−13 Two-photon (1,040 nm) fluorescence in live HEK cells with ionomycin jRGECO1a versus RCaMP3: P = 6.9 × 10−14; XCaMP-R versus RCaMP3: P = 6.9 × 10−14 Ca2+ imaging with whole-cell patch-clamp recording in acute brain slices Representative jRGECO1a and RCaMP3 responses to single action potentials (APs Thin lines denote individual traces (jRGECO1a: 12 trials rise time (g) and half-decay time (h) for single APs P = 4.7 × 10−2 (f); P = 9.7 × 10−1 (g); P = 4.6 × 10−1 (h) Ca2+ imaging under a cell-attached recording in vivo Representative trace of simultaneous measurement of RCaMP3 fluorescence and APs in vivo The number of APs for each event is indicated below the trace The image shows a neuron expressing RCaMP3 with the recording pipette ΔF/F of jRGECO1a (152 events from 12 cells) and RCaMP3 (228 events from 15 cells) for single APs and thick lines represent the average traces Half-rise time (l) and half-decay time (m) for single APs P = 4.5 × 10−58 (1AP); P = 1.8 × 10−93 (2AP); P = 1.2 × 10−60 (3AP); P = 3.8 × 10−17 (4AP) magnified images and Ca2+ traces of 12 representative neurons Source data AAVs encoding RCaMP3 and cAMPinG1-ST were co-injected into the L2/3 of the V1 Sequential excitation at 940 nm and 1,040 nm was used for dual-color imaging of cAMPinG1-ST and RCaMP3 Three optical planes spaced 30 µm apart were imaged at 3.4 Hz per plane using a piezo objective scanner Representative images of RCaMP3 and cAMPinG1-ST Single-trial traces of RCaMP3 and cAMPinG1-ST Cells are sorted according to ΔF/F of RCaMP3 during running Single-trial traces of RCaMP3 and cAMPinG1-ST of two representative cells The box indicates the period of forced running The cell number on the top corresponds to the number in d Averaged fluorescence transients of RCaMP3 (magenta) and cAMPinG1-ST (green) Cumulative plot of mean cAMP ΔF/F of motion-related (green) and non-related (black) cells during forced running Averaged ΔF/F of RCaMP3 (magenta) and cAMPinG1-ST (green) during forced running Source data Moving gratings of 8 directions were used to induce cell-specific Ca2+ transients in L2/3 neurons of the V1 ΔF/F of RCaMP3 and cAMPinG1-ST of 2 representative cells Direction-selective visual responses of RCaMP3 and cAMPinG1-ST of the two representative cells in b Direction-selective visual responses of RCaMP3 and cAMPinG1-ST neurons showing the direction selectivity index (DSI) < 0.4 in Ca2+ response neurons showing the DSI ≥ 0.4 in Ca2+ response Single-trial traces of RCaMP3 and cAMPinG1-ST of 2 representative cells The box indicates the period of visual stimuli Averaged traces of RCaMP3 (magenta) and cAMPinG1-ST (green) ΔF/F of RCaMP3 and cAMPinG1-ST during and after the visual stimuli Responded (during stim) versus non-responded (during stim): P = 1.5 × 10−7; responded (during stim) versus responded (after stim): P = 2.1 × 10−4; non-responded (during stim) versus non-responded (after stim): P = 5.1 × 10−4 Schematic of AAVs for sparse expression of cAMPinG1-NE and soma-targeted ChRmine Representative fluorescence images of cAMPinG1-NE and soma-targeted ChRmine ΔF/F of cAMPinG1-NE in response to 1,040 nm of photostimulation photostim (+): P = 1.4 × 10−2; ChRmine (+) Source data two-photon imaging of RCaMP3 and cAMPinG1-ST revealed bidirectional cAMP change and strong correlation between Ca2+ and cAMP with single-cell resolution in vivo indicating that action potentials are sufficient to induce somatic cAMP elevation These fluorescence signals arose from cAMP binding and were more intense than those of G-Flamp1 Representative images of the cAMPinG1 stable cell line transiently expressing DRD1–P2A–RFP in the absence (middle) or presence (bottom) of 1,000 nM dopamine were taken by alternating 405/488/561-nm lasers Correlation between 488 ex/405 ex ratio of cAMPinG1 and DRD1–P2A–RFP expression level Pearson correlation coefficient in linear regression r = 0.66; P = 2.7 × 10−19 (top) and r = −0.03; P = 0.64 (bottom) Representative images of cAMPinG1 cells expressing DRD2–P2A–RFP in the absence (middle) or presence (bottom) of 1,000 nM dopamine Correlation between 488 ex/405 ex ratio and DRD2–P2A–RFP expression level r = 0.07; P = 0.35 (top) and r = −0.59; P = 4.9 × 10−16 (bottom) 488 ex/405 ex ratio of cAMPinG1 cells expressing GPCRs–P2A–RFP without ligands 178 (GPR52) and 126 (no transfection) cells ARDB2 versus DRD2: P = 9.7 × 10−14; DRD1 versus DRD2: P = 9.7 × 10−14; ARDB2 versus GPR52: P = 9.7 × 10−14; DRD1 versus GPR52: P = 9.7 × 10−14 488 ex/405 ex ratio of cAMPinG1 cells expressing GPCRs–P2A–RFP with or without agonists 180 (FSK + DRD2) and 154 (dopamine + FSK + DRD2) cells (left) DRD1: P = 6.3 × 10−113; DRD2: P = 2.7 × 10−69 Tukey’s post hoc test following one-way ANOVA (right) No drug versus serotonin: P = 5.4 × 10−5; no drug versus clozapine: P = 1.5 × 10−14; serotonin versus clozapine: P = 1.5 × 10−14 Source data a multicolor suite for cAMP and Ca2+ imaging and showed applications that required advanced sensitivity of these indicators we addressed an important biological question: the relation between Ca2+ and cAMP signaling in vivo The detection of these bidirectional cAMP changes indicates that the cAMP affinity of cAMPinG1 is suitable for in vivo cAMP imaging while the other non-cell-autonomous mechanisms cannot be ruled out Our results suggest the possibility that cAMP can encode specific information such as direction selectivity in vision or locomotion encoded in action potentials and Ca2+ signaling This cell-specific cAMP elevation cooperated or competed with global cAMP increase or decrease leading to the formation of population patterns of cAMP much longer than the hundreds of milliseconds of Ca2+ transients information encoded in Ca2+ and GPCR signaling can be integrated and stored for a longer period through cAMP transients All experimental procedures were performed using protocols that were approved by the Institutional Animal Care and Use Committee at Kyoto University (Lif-K23008) RIKEN (W2022-2-012) and The University of Tokyo (P18-118) ICR) and A2A-Cre ((B6.FVB (Cg)-Tg (Adora2a-cre) KG139Gsat/Mmucd)) mice were group-housed and kept on a 12-h light–dark cycle with ad libitum food and water The housing conditions were controlled at room temperature of approximately 22–24 °C and a relative humidity of 40–60% Wild-type mice used in this study were purchased from Japan SLC Experiments were performed using both male and female sex between 0 and 20 weeks of age HEK293T cells were obtained from the American Type Culture Collection (CRL-11268) Nacalai) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich) 50 units per ml penicillin and 50 μg ml−1 streptomycin (26252-94 Nacalai) at 37 °C and 5% CO2 in a humidified atmosphere and Stbl3 cells were obtained from Toyobo (DNA-9303) Invitrogen (18297010) and Invitrogen (C737303) Bacteria were incubated in Lysogeny Broth (LB) medium supplemented with antibiotics at 37 °C The plasmids for bacterial expression of cAMP sensors were transformed into E coli cells were plated and cultured at 37 °C on an LB agar plate with ampicillin and 0.0004% arabinose Each colony was used to inoculate 1.5 ml of LB liquid medium with ampicillin and 0.2% arabinose and grown at 37 °C overnight cells were resuspended in 150 µl suspension buffer (20 mM MOPS (pH 7.2) cOmplete EDTA-free protease inhibitor cocktail; 11836170001 the supernatant was diluted 20-fold with the suspension buffer The diluted supernatant was applied to 96-well plates Tokyo Chemical Industry) was added to a final concentration of 300 µM for cAMP-saturated conditions Fluorometric measurements were performed on a microplate reader (Spark TECAN) at room temperature at 485 nm of excitation (bandwidth of 20 nm) and 535 nm of emission (bandwidth 20 nm) and cAMP was added to a final concentration of 100 µM pKa values were determined from the inflection point of a sigmoid fit to fluorescence versus pH The plasmids for bacterial expression of cAMPinG1 coli cells were plated and cultured at 37 °C on an agar plate with ampicillin A colony was used to inoculate 200 ml of LB liquid medium with ampicillin and 0.2% arabinose and grown at 18 °C for 44 h cells were resuspended in 10 ml suspension buffer (25 mM Tris-HCl (pH 8.0) 1 mM DTT and cOmplete EDTA free (Sigma-Aldrich)) cAMP was microperfused from 0.5 μM to 0 μM 256 × 48 pixels) were collected at 12.7 Hz For time-lapse imaging (Fig. 1h–j) HEK293T cells were incubated in 35-mm glass-bottom dishes DNA encoding the sensors (1 µg) was transfected as described above the culture medium was replaced with Tyrode’s solution (129 mM NaCl Imaging for cAMPinG1 was performed using an LSM 880 confocal microscope (Carl Zeiss) with an air-immersion ×20 objective lens (N.A.: 0.80 405-nm and 488-nm lasers were used for excitation in turns Nacalai) was added to a final concentration of 50 µM For single-timepoint imaging for GPCRs (Fig. 6) HEK293T cells were incubated in 96-well glass-bottom plates Around 0.1 µg DNA encoding the sensors was transfected as described above the culture medium was replaced with Tyrode’s solution Imaging was performed using an LSM 880 confocal microscope with the ×20 objective lens For tetracycline-dependent expression of GPCRs–P2A–mCherry doxycycline was added to a final concentration of 100 ng ml−1 3 h before the imaging the culture medium was replaced with Tyrode’s solution with or without forskolin Lasers (405 nm and 488 nm) were used for ratiometric cAMP imaging and 561-mn and 633-nm lasers were used for the visualization of GPCR-expressing cells For single-timepoint imaging for sensor comparison (Extended Data Fig. 9a–c) Imaging was performed to calculate the relative change (ΔR/R) in fluorescence ratio (R) in the absence or presence of 10 μM forskolin R is the ratio of green fluorescence (491–553 nm of emission) with 488 nm of excitation to that with 405 nm of excitation R is the ratio of cyan fluorescence (464–499 nm of emission) with 458 nm of excitation to yellow fluorescence (526–597 nm of emission) with 514 nm of excitation the culture medium was replaced with Tyrode’s solution as described above Imaging was performed using an LSM 880 confocal microscope with an oil-immersion ×40 objective lens (N.A.: 1.30 512 × 512 pixels) were collected using 488-nm and 561-nm lasers HEK293T cells in 96-well glass-bottom plates were prepared as described above the culture medium was replaced with Tyrode’s solution with or without 10 µM forskolin Imaging was performed using a TCS SP8 FALCON microscope (Leica) at a pulse frequency of 80 MHz with an air-immersion ×20 objective lens (N.A.: 0.75 Excitation was 488 nm by white-light laser and emission was 500–550 nm The lifetime was analyzed using the LAS X FLIM/FCS software (Leica) A mixture of 0.8 µg DNA encoding the red Ca2+ sensors and 0.2 µg DNA of pCMV-mCerulean was transfected as described above Thirty seconds after bath application of ionomycin (Cayman Chemical two-photon imaging was performed with an FVMPE-RS (Olympus) equipped with a water-immersion ×25 objective lens (N.A.: 1.05 Spectra-Physics) and two GaAsP detectors (Hamamatsu Photonics) with 495–540-nm and 575–645-nm emission filters (Olympus) The laser was tuned to 880 nm for mCerulean and 1,040 nm at the front aperture of the objective for the red Ca2+ sensors imaging was performed using a FVMPE-RS microscope with the ×20 objective lens in Tyrode’s solution 256 × 256 pixels) were collected at 0.82 Hz The HEK293T cells were seeded on six-well plates with 2 ml DMEM and 10% FBS or 1.5% FBS and incubated at 37 °C The 1.5% FBS group was a positive control of slow cell proliferation Twenty hours after the beginning of the culture and the number of cells was counted using a counting chamber as a timepoint of zero Sixty hours after the beginning of the culture cells in the other half of the wells were harvested and counted as a timepoint of 48 h Electroporated mice were used for cAMP and calcium imaging 4–10 weeks after birth Stereotaxic virus injection was performed to C57BL/6N male mice aged 4–6 weeks anesthetized by the anesthetic mixture described above except for two-photon mesoscale imaging. A2A-Cre transgenic mice were used for the slice experiments for local dopamine application (Extended Data Fig. 3e–h) A micropipette was inserted into the right V1 (A/P −3.85 mm D/V −4.5 mm) or the medial prefrontal cortex (A/P +1.8 mm the virus solution (volume: 500−1,000 nl) was injected Zoetis) was administered intraperitoneally just after the injection experiment Mice were subjected to imaging after 4–12 weeks of the injection 1 µM dopamine was microperfused with a micropipette For cAMP imaging with norepinephrine bath application (Extended Data Fig. 3i–l) in utero electroporation was performed as described above mice were killed by rapid decapitation after anesthesia with the anesthetic mixture described above The brains were immediately extracted and immersed in gassed (95% O2/5% CO2) and ice-cold solution containing: 222 mM sucrose Acute coronal brain slices (300 μm thick) of the visual cortex were cut in gassed ice-cold solution with a vibratome (VT1200 Brain slices were then transferred to an incubation chamber containing gassed ACSF containing: 126 mM NaCl 2 mM MgCl2 and 10 mM glucose at room temperature for 30 min The brain slices were transferred to the recording chamber and perfused with the ACSF solution described above cAMP imaging was performed with FVMPE-RS (Olympus) equipped with a water-immersion ×25 objective lens (N.A.: 1.05 Spectra-Physics) and two GaAsP detectors (Hamamatsu Photonics) with 495–540-nm emission filters (Olympus) 256 × 256 pixels) with 36 optical planes with planes spaced 2 µm apart in depth were collected every 31 s Tokyo Chemical Industry) was added to a final concentration of 0.5 µM For characterization of cAMPinG1 in acute brain slices (Extended Data Fig. 3m–o) AAVPHP.eB-hSyn-EGFP) was injected into the medial prefrontal cortex of mice aged 8 weeks at a volume of 500 nl and acute brain slices were prepared as described above Slices were perfused with oxygenated ACSF (125 mM NaCl Whole-cell recordings were performed by 5–6 MΩ glass pipettes Patch pipettes were filled with an internal solution (120 mM potassium gluconate Electrophysiological data were acquired using a patch-clamp amplifier (MultiClamp 700B Molecular devices) and stored using a Digidata 1440A converter and pCLAMP software (Molecular Devices) the spike number was measured by injecting pulses of increased intensity in steps of 25 pA (from 0 to 250 pA For miniature excitatory postsynaptic current measurement tetrodotoxin (0.2 μM to a final concentration) APV (50 μM) and picrotoxin (25 μM) were added and membrane potential voltage clamped at −70 mV after correction of liquid-junction potential was used Tissue blocks were cut into 50-μm-thick slices with a cryostat (CM1950 brain slices were incubated with DAPI for 10 min at room temperature before being mounted on slides Imaging was performed using an LSM 880 confocal microscope with a ×20 objective lens and 405-nm 512 × 512 pixels) were acquired across multiple optical planes each spaced 2 µm apart in depth from L2/3 neurons in the V1 AAV2/1-eSyn-RCaMP3) was injected into the barrel cortex (A/P −1.0 mm D/V −0.2 mm from the pial surface) at 20 nl min−1 at a volume of 500 nl mice were killed by rapid decapitation after anesthesia with pentobarbital (100 mg per kg body weight) The brains were immediately extracted and immersed in gassed (95% O2/5% CO2) and ice-cold ACSF containing: 124 mM NaCl Acute coronal brain slices (300 μm thick) of the barrel cortex were cut in gassed Brain slices were then transferred to an incubation chamber containing gassed ACSF at 30 °C for 60 min and subsequently maintained at room temperature before transferring them to the recording chamber at 35 °C Whole-cell recordings were performed in the L2/3 pyramidal neurons of the barrel cortex with glass recording electrodes (5–8 MΩ) filled with the intracellular solution containing: 130 mM K-gluconate 7 mM dipotassium-phosphocreatine and pH adjusted to 7.0 with potassium hydroxide (296 mOsm) Molecular devices) filtered at 10 kHz and sampled at 20 kHz Single action potentials were evoked by injecting a series of current pulses (2 ms in duration) through the patch pipette Calcium imaging was performed using an upright microscope (BX51WI Olympus) with a water-immersion ×40 objective lens (N.A.: 0.8 To acquire RCaMP3 and jRGECO1a images with LED light (MCWHLP1 a U-MWIG3 fluorescence mirror unit (Olympus) was used Fluorescent images were captured by a sCMOS camera (Orca-Flash 4.0 v3 Hamamatsu Photonics) controlled by HC Image software (Hamamatsu Photonics) Images were acquired at 50 Hz with 1 × 1 binning mice were head-fixed and anesthetized with isoflurane (~1.5–2.0%) throughout the experiment and body temperature was kept at 37 °C with a heating pad A craniotomy was made in the barrel cortex The exposed brain was covered with 1.5% agarose in ACSF containing the following: 150 mM NaCl A glass coverslip was then placed over the agarose to suppress the brain motion artifacts A glass electrode (5–8 MΩ) was filled with ACSF containing Alexa 488 (200 µM) jRGECO1a or RCaMP3-expressing neurons were targeted using two-photon microscopy (Movable Objective Microscope Spectra-Physics) and a water-immersion ×16 objective lens (N.A.: 0.80 Fluorescence signals were collected using a GaAsP photomultiplier tube (Hamamatsu Photonics) with a 590–660-nm emission filter After establishing the cell-attached configuration (20–100 MΩ seal) simultaneous spike recording and calcium imaging were performed at the soma (sampling rate = 30 Hz Electrophysiological data were acquired using a patch-clamp amplifier (MultiClamp 700B; Molecular devices) in current-clamp mode The laser was tuned to 1,040 nm (40 mW at the front aperture of the objective) A 400-µm-diameter mono fiber-optic cannula (Kyocera) was implanted A custom-made metal head plate was attached to the skull with dental cement Mice were subjected to imaging after more than 2 days of the surgery Dual-color fiber photometry for green cAMP sensors and RCaMP3 was performed using the GCaMP and Red Fluorophore Fiber Photometry System (Doric) with 405-nm Photometry data were recorded at a sampling rate of 30 Hz by lock-in amplifier detection Mice were head-fixed during the recordings just after the first recording for 30-s forced running 25 mg per kg body weight propranolol (168-28071 Wako) or mock solution was intraperitoneally administered the same two recordings were performed for the same mice Half of the mice received propranolol on the first day and the other half received mock solution on the first day The mice were subjected to imaging more than 18 h after the surgery 192 × 256 pixels) were collected at 1.6 Hz in the awake condition The laser power for 940 nm of excitation was set to 26.5 mW For single-cell cAMP imaging with optical stimulation using soma-targeted ChRmine 192 × 256 pixels) were collected at 3.2 Hz in the awake condition with 940 nm of excitation 1,040-nm two-photon excitation was used for 4-s The imaging with a 940-nm laser was temporally stopped during the optical excitation The laser power for 940-nm excitation was set to 4.1 mW AAV (AAV2/1-eSyn-RCaMP3) was injected into the neonatal somatosensory cortex53 a 4.5-mm-diameter craniotomy was performed over an area including the primary somatosensory area of the right hemisphere A head plate was also fixed to the skull above the cerebellum The field of view was 3.0 × 3.0 mm2 (2,048 × 2,048 pixels) Laser power of 270 mW and 360 mW at the front of the objective lens was used to observe L2/3 and L5 neurons of awake mice The gratings were presented with an LCD monitor (19.5 inches placed 25 cm in front of the center of the left eye of the mouse Each stimulus trial consisted of a 4-s blank period (uniform gray at mean luminance) followed by a 4-s drifting sinusoidal grating (0.04 cycles per degree Eight drifting directions (separated by 45° The timing of each moving grating stimulus and the initiation of imaging were monitored with a data acquisition module (USB-6343 National Instruments) controlled by LabVIEW (2021) half-rise time and half-decay time were calculated by single exponential fitting No fluorescence cross-talk correction was performed where F is the fluorescence intensity at any timepoint and F0 is the average fluorescence before the drug application For in vivo cAMPinG1-NE spine imaging (Fig. 2g–i) the period after stimulation was defined as a 15-s period starting 10 s after the end of airpuff stimulation For two-photon Ca2+ imaging in HEK293T cells (Fig. 3c) ROIs drawn based on mCerulean images with Cellpose were used for both Ca2+ sensor and mCerulean images The red fluorescence intensity was divided by mCerulean fluorescence intensity in each cell for normalization For Ca2+ imaging using acute brain slices (Fig. 3d–h) background subtraction and bleach correction were performed before calculating ΔF/F ROIs were manually selected around somata in the time-series-averaged image where F is the fluorescence intensity at any timepoint and F0 is the resting baseline fluorescence measured 200 ms before stimulation The peak amplitude was defined as the maximum value of ΔF/F after the stimuli The rise and decay curves were fit to a single exponential The rise time was defined as the time from the beginning of the stimulus to the timepoint of the peak fluorescence amplitude The half-decay time was defined as the time from the maximum value of ΔF/F to half of that value For simultaneous Ca2+ imaging and cell-attached recordings in vivo (Fig. 3i–n) where F is the fluorescence intensity at any timepoint and F0 is resting baseline fluorescence measured 200 ms before the action potentials Action potentials were detected by cell-attached recording of the signal Spike events (1–4 action potentials) were identified ensuring that no other action potentials occurred in the 1-s period before and after the first action potential The half-rise time was defined as the time from the beginning of the stimulus to the timepoint of the half of the peak fluorescence amplitude The period after repetitive visual stimuli was defined as a 40-s period starting after the end of the stimuli The DSI was calculated for cells showing Ca2+ responses The preferred direction (θpref) of each cell was defined as the stimulus that induced the largest Ca2+ ΔF/F The DSI was defined as DSI = (Rpref – Rpref+π)/(Rpref + Rpref+π) where Rpref and Rpref+π are ΔF/F values at the preferred (θpref) and the opposite (θpref + π) directions Imaging frames with notable motion artifacts were removed and supplied with the preceding frames For cAMPinG1-NE imaging with soma-targeted ChRmine (Fig. 5k) cAMP ΔF/F was defined as averaged ΔF/F during a 10-s period starting 20 s after the end of optical stimulation Fiber photometry analysis was performed using Python The cAMPinG1 signal was calculated as follows: (470-nm signal)/(405-nm signal) The 560-nm signal was recognized as the RCaMP3 signal For imaging in the V1 (Extended Data Fig. 6e–g) where F is mean fluorescence intensity in the last 10 s of the running period and F0 is the mean fluorescence intensity in 10 s before the start of running ΔR/R (pre) and ΔR/R (post) were defined as ΔR/R in response to running before and after the intraperitoneal administration For imaging in the dorsal striatum (Extended Data Fig. 8) the period during stimulation was defined as a 10-s running period and the period after stimulation was defined as 10 s after the end of the running period All experiments were conducted with at least two biological replicates involving independent transfection and mice All statistical analyses of the acquired data were performed with Python a statistical test matching the structure of the experiment and the structure of the data was employed *P < 0.05; **P < 0.01; ***P < 0.001; NS not significant (P > 0.05) for all statistical analyses presented in figures No statistical tests were done to predetermine the sample size Data acquirement and analysis were not performed blind to the conditions of the experiments Experimental sample sizes are mentioned in the figure panel and legends Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article The molecular and systems biology of memory Fluorescence ratio imaging of cyclic AMP in single cells Genetically encoded sensors towards imaging cAMP and PKA activity in vivo Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging Distinct temporal integration of noradrenaline signaling by astrocytic second messengers during vigilance Hypothalamic dopamine neurons motivate mating through persistent cAMP signalling A high-performance genetically encoded fluorescent indicator for in vivo cAMP imaging Sensitive genetically encoded sensors for population and subcellular imaging of cAMP in vivo Green fluorescent cAMP indicator of high speed and specificity suitable for neuronal live-cell imaging An improved genetically encoded fluorescent camp indicator for sensitive cAMP imaging and fast DRUG SCreening New DAG and cAMP sensors optimized for live-cell assays in automated laboratories A family of hyperpolarization-activated mammalian cation channels Structure and allostery of the PKA RII tetrameric holoenzyme Subcellular dynamics of Type II PKA in neurons Classification and phylogenetic analysis of the cAMP-dependent protein kinase regulatory subunit family PKA-I holoenzyme structure reveals a mechanism for cAMP-dependent activation Regulatory subunit of protein kinase A: structure of deletion mutant with cAMP binding domains Mutations of PKA cyclic nucleotide-binding domains reveal novel aspects of cyclic nucleotide selectivity RIalpha subunit of PKA: a cAMP-free structure reveals a hydrophobic capping mechanism for docking cAMP into site B Engineering extrinsic disorder to control protein activity in living cells An improved genetically encoded red fluorescent Ca2+ indicator for detecting optically evoked action potentials Soma-targeted imaging of neural circuits by ribosome tethering Genetically-encoded yellow fluorescent cAMP indicator with an expanded dynamic range for dual-color imaging a multicolor GECI suite for in vivo imaging of complex brain circuit dynamics Sensitive red protein calcium indicators for imaging neural activity An expanded palette of genetically encoded Ca2+ indicators Improved orange and red Ca2+ indicators and photophysical considerations for optogenetic applications contiguous-wide two-photon imaging to reveal functional network architectures across multi-modal cortical areas A highly sensitive a-kinase activity reporter for imaging neuromodulatory events in awake mice Pupil fluctuations track rapid changes in adrenergic and cholinergic activity in cortex Spontaneous behaviors drive multidimensional Astrocyte heterogeneity revealed by expression of a GFAP-LacZ transgene Ultrasensitive fluorescent proteins for imaging neuronal activity Cortical layer-specific critical dynamics triggering perception Diametric neural ensemble dynamics in parkinsonian and dyskinetic states Constitutive activity of a G protein-coupled receptor contributes to human cerebral organoid formation Structural basis of ligand recognition and self-activation of orphan GPR52 Imaging cytoplasmic cAMP in mouse brainstem neurons Phase separation of a PKA regulatory subunit controls cAMP compartmentation and oncogenic signaling Receptor-associated independent cAMP nanodomains mediate spatiotemporal specificity of GPCR signaling A critical time window for dopamine actions on the structural plasticity of dendritic spines Common coupling map advances GPCR-G protein selectivity A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein eqFP578 PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome Oscillatory control of factors determining multipotency and fate in mouse neural progenitors Near-infrared fluorescent proteins for multicolor in vivo imaging In utero electroporation and cranial window implantation for in vivo wide-field two-photon calcium imaging using G-CaMP9a transgenic mice A Flp-dependent G-CaMP9a transgenic mouse for neuronal imaging in vivo Functional labeling of neurons and their projections using the synthetic activity-dependent promoter E-SARE Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems Protocol for cortical-wide field-of-view two-photon imaging with quick neonatal adeno-associated virus injection Pachitariu, M. et al. Suite2p: beyond 10,000 neurons with standard two-photon microscopy. Preprint at bioRxiv https://doi.org/10.1101/061507 (2016) A pyramid approach to subpixel registration based on intensity Cellpose: a generalist algorithm for cellular segmentation Download references We thank the following researchers for kindly sharing their reagents: H and a protocol about kinetics analysis); T Massachusetts Institute of Technology (Addgene plasmid Albert Einstein College of Medicine (Addgene plasmid Korea Institute of Science and Technology (Addgene plasmid Tokyo Institute of Technology (Addgene plasmid California Institute of Technology (Addgene plasmid University of North Carolina at Chapel Hill (Addgene kit Kato for technical assistance and members of the laboratories of M.S This work was supported in part by grants from Precursory Research for Embryonic Science and Technology (PRESTO)-JST (JPMJPR1906 to M.S.) Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) (JP19dm0207079 to S.M. Interdisciplinary and Emerging Brain Research Program (iBrain/MINDS) (JP21wm0525018 to S.Y Program for Technological Innovation of Regenerative Medicine (JP21bm0704060 to I.I.) Interstellar Initiative Beyond (JP22jm0610068 to M.S.) RIKEN Special Postdoctoral Researchers Program (to H.U.) The Konica Minolta Science and Technology Foundation (to M.S.) Tokyo Biochemical Research Foundation (to M.S.) Mochida Memorial Foundation (to M.S.) and KOSÉ Cosmetology Research Foundation (to M.S.) This work was also supported by Kyoto University Live Imaging Center Department of Optical Neural and Molecular Physiology Center for Living Systems Information Science Department of Brain Development and Regeneration Laboratory of Deconstruction of Stem Cells Institute for Frontier Life and Medical Sciences Laboratory for Haptic Perception and Cognitive Physiology Precursory Research for Embryonic Science and Technology (PRESTO) performed most of the experiments and analyzed data performed electrophysiological recordings for RCaMP3 characterization performed Ca2+ imaging with the FASHIO-2PM performed electrophysiological and pharmacological experiments for cAMPinG1 in acute brain slices designed the study and wrote the paper with input from all authors The remaining authors declare no competing interests Nature Methods thanks Xiaoke Bi and the other reviewer(s) for their contribution to the peer review of this work in collaboration with the Nature Methods team Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Note the cpGFP insertion site is close to cAMP in the cAMP-bound three-dimensional structure and exposed on the surface The amino acid sequence of cAMP-binding domains of PKA regulatory-subunit Excitation and emission spectra of cAMP-free and cAMP-saturated G-Flamp1 Fluorescence intensities of cAMP sensors in cAMP-free and cAMP-saturated states in HEK293T cell lysate gCarvi vs cAMPinG1: P = 4.2 × 10−13; G-Flamp1 vs cAMPinG1: P = 2.4 × 10−2 Flamindo2 vs gCarvi: P = 6.3 × 10−4; gCarvi vs G-Flamp1: P = 1.3 × 10−3; gCarvi vs cAMPinG1: P = 2.1 × 10−3 cGMP and cAMP titration curves of Flamindo2 and cAMPinG1 pH titration of cAMP-free and cAMP-saturated cAMP sensors In vitro binding kinetics of cAMPinG1 in response to cAMP microperfusion from 0 μM to 2 μM In vitro dissociation kinetics of cAMPinG1 in response to cAMP microperfusion from 0.5 μM to 0 μM Representative images of binding assay of PKA-R1α-mEGFP and cAMPinG1 with PKA-catalytic subunit (Cat)-RFP-CAAX in HEK293T cells Lines are for quantification of fluorescence distribution for (j) Quantification of line fluorescence distribution as detected by the red fluorescence of PKA-Cat-RFP-CAAX Fluorescence lifetime of cAMP sensors and fluorescence proteins expressed in HEK293T cells cAMPinG1 in the absence vs presence of 10 μM forskolin: P = 1.8 × 10−5; G-Flamp1 in the absence vs presence of 10 μM forskolin: P = 8.5 × 10−2 All shaded areas and error bars denote SEM Source data Traces of cAMPinG1-NE (left) and cAMPinG1mut-NE (right) in response to 25 µM forskolin and 50 µM IBMX and colored thick lines denote average response ΔF/F for the forskolin (FSK) and IBMX application n = 12 neurons in 2 slices (cAMPinG1mut-NE) Schematic of “Cre-Off” system for expressing cAMP sensors in all neurons and mCherry in dopamine receptor D1-stratal projection neurons (D1R-SPNs) in the striatum Schematic of imaging settings (left) and representative images of cAMPinG1-NE in an acute brain slice (right) Fluorescence in response to 10 s local puff of 1 µM dopamine was recorded in mCherry-positive D1R-SPNs (arrow) Dopamine receptor D2 (D2R)-positive neurons were not labeled with mCherry (arrowhead) due to the expression of Cre recombinase The responses to dopamine were aligned with a single-exponential fitting n = 7 neurons in 3 slices (cAMPinG1-NE to dopamine) n = 11 neurons in 4 slices (cAMPinG1-NE to ACSF) n = 9 neurons in 4 slices (G-Flamp1 to dopamine) Schematic of imaging settings (left) and representative images (right) Traces of cAMPinG1-NE and G-Flamp1 in response to 0.5 µM noradrenaline Number of spikes evoked by current injection in expressing cAMPinG1-NE and EGFP neurons mEPSC (miniature excitatory postsynaptic current) frequency (left) and amplitude (right) in cAMPinG1-NE and EGFP expressing neurons Frequency: P = 2.5 × 10−1; Amplitude: P = 4.9 × 10−1 Source data Mutations are indicated in R-GECO1 numbering Fluorescence intensities of red Ca2+ sensors in Ca2+-free and Ca2+-saturated conditions in HEK293T cell lysate Ca2+-free jRGECO1a vs Ca2+-free RCaMP3: P = 2.6 × 10−5; Ca2+-free XCaMP-R vs Ca2+-free RCaMP3: P = 1.9 × 10−7 Ca2+-saturated jRGECO1a vs Ca2+-saturated RCaMP3: P = 1.3 × 10−9; Ca2+-saturated XCaMP-R vs Ca2+-saturated RCaMP3: P = 1.4 × 10−12 Excitation and emission spectra of jRGECO1a and RCaMP3 in Ca2+-free and Ca2+-saturated states Comparison of excitation spectra of red Ca2+ indicators and Hill coefficients (i) of red Ca2+ sensors Kd values (h) of jRGECO1a vs RCaMP3: P = 9.0 × 10−1; XCaMP-R vs RCaMP3: P = 8.0 × 10−6 Hill coefficients (i) of jRGECO1a vs RCaMP3: P = 3.8 × 10−2; XCaMP-R vs RCaMP3: P = 6.1 × 10−8 Normalized fluorescence of red Ca2+ sensors in Ca2+-free and Ca2+-saturated states as a function of pH One-photon (k) and two-photon (l) bleaching curves of jRGECO1a XCaMP-R and RCaMP3 expressed in HEK293T cells Source data Schematic of the experimental procedure of two-photon mesoscale Ca2+ imaging using fast-scanning high optical invariant two-photon microscopy (FASHIO-2PM) Left: A representative full FOV of FASHIO-2PM Right: Magnified images and Ca2+ traces of representative 6 somata and 6 dendrites Representative two-photon images and averaged traces of cAMPinG1-ST (a) and G-Flamp1 (c) in response to forced running and G-Flamp1 in response to forced running cAMPinG1-ST vs cAMPinG1mut-ST: P = 2.6 × 10−2 cAMPinG1-NE was expressed in neurons in the V1 Fiber photometry was performed during forced running Thirty minutes after intraperitoneal administration of propranolol fiber photometry was again performed during the forced running Averaged traces of cAMPinG1-NE in response to forced running before and after intraperitoneal administration of mock solution (left) or propranolol (right) Difference of ΔF/F in response to forced running before and after the intraperitoneal administration Representative images of cAMPinG1 and RCaMP3 Averaged fluorescence transients of cAMPinG1 and RCaMP3 Source data a,b, Representative two-photon images and traces of RCaMP3 and cAMPinG1-ST of the cells in Fig. 5b-c and G-Flamp1 (right) in response to visual stimuli and G-Flamp1 in response to visual stimuli Unpaired two-tailed t-test after removing outliers with Smirnov-Grubbs’ test (P = 0.05) cAMPinG1-ST vs cAMPinG1mut-ST: P = 1.9 × 10−5 Averaged ΔF/F of RCaMP3 and cAMPinG1mut-ST in cells which showed Ca2+ response to visual stimuli Source data AAVs encoding RCaMP3 and green cAMP sensors were injected into the dorsal striatum (dStr) Dual-color fiber photometry was performed in the dStr during a forced running task we employed different excitation wavelengths: 560 nm for RCaMP3 imaging and 405 nm and 470 nm for cAMPinG1 ratiometric imaging Representative single-trial traces of cAMPinG1-NE (left) Averaged fluorescence traces of cAMPinG1-NE (left) cAMPinG1mut-NE and G-Flamp1 during and after the stimulation cAMPinG1-NE vs cAMPinG1mut-NE (during stim): P = 3.7 × 10−4; cAMPinG1-NE vs G-Flamp1 (during stim): P = 3.6 × 10−5; cAMPinG1-NE vs cAMPinG1mut-NE (after stim): P = 3.1 × 10−5; cAMPinG1-NE vs G-Flamp1 (after stim): P = 4.2 × 10−5 Source data Single timepoint imaging of cAMPinG1 (top) and G-Flamp1 (bottom) in the absence or presence of 10 μM forskolin Single timepoint imaging of cAMPFIRE-L in the absence or presence of 10 μM forskolin Relative change (ΔR/R) in fluorescence ratio (R) in the absence or presence of 10 μM forskolin R is the ratio of green fluorescence with 488 nm excitation to that with 405 nm excitation R is the ratio of cyan fluorescence with 458 nm excitation to yellow fluorescence with 514 nm excitation cAMPinG1 vs G-Flamp1: P < 1.0 × 10−13; cAMPinG1 vs cAMPFIRE-L: P < 1.0 × 10−13; G-Flamp1 vs cAMPFIRE-L: P < 1.0 × 10−13 A representative image of the cAMPinG1 stable cell line Proliferation assay of the cAMPinG1 stable cell line and original HEK293T cell line The original HEK cell line in 1.5 % FBS condition was for a negative control of the assay cAMPinG1 cell line in 10% FBS vs original cell line in 1.5% FBS: P = 1.9 × 10−5; Original cell line in 10% FBS vs original cell line in 1.5% FBS: P = 5.0 × 10−4 Single timepoint imaging of cAMPinG1 cell line in the absence (top) or presence (bottom) of 50 μM forskolin Source data and mCherry triple stable cell line in the absence (top) and presence (bottom) of 100 nM dopamine a Gs-coupling GPCR) and iRFP670 triple stable cell line in the absence (top) or presence (bottom) of 1,000 nM ACTH Representative images of a mixture of the cell lines with 100 nM dopamine (middle) or 1,000 nM ACTH (bottom) Representative cells expressing DRD1/RFP or MC3R/iRFP were indicated by arrows or arrowheads 488 ex/405 ex ratio of each cell line in the presence of dopamine or ACTH ACTH + DRD1 vs ACTH + MC3R: P = 2.4 × 10−11; ACTH + DRD1 vs dopamine + DRD1: P = 2.1 × 10−13 ACTH + MC3R vs dopamine + MC3R: P = 2.1 × 10−13; dopamine + DRD1 vs dopamine + MC3R: P = 2.1 × 10−13 Source data FASHIO-2PM for RCaMP3 imaging from L5 neurons in an awake condition The size of the imaging area is 3.0 × 3.0 mm2 (2,048 × 2,048 pixels) Images are filtered with Kalman stack filter by ImageJ Mouse was presented with moving grating in eight directions to the contralateral eye in awake condition Sequential excitation at 940 nm and 1,040 nm was used for dual-color imaging of RCaMP3 and cAMPinG1-ST Three optical planes spaced 30 μm apart were imaged at 3.4 Hz per plane using a piezo objective scanner The size of the imaging area is 339 × 339 μm2 (512 × 512 pixels) Download citation DOI: https://doi.org/10.1038/s41592-024-02222-9 Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. 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Brand: SmackdownHeight: 5'7"Weight: 205 lbsHometown: Tokyo JapanFinishing Move: Kick of DeathOther Aliases: Yoshihiro Tajiri Yoshihiro Tajiri is one of the most respected athletes in the WWE locker room Beginning his career for the International Wrestling Association Tajiri became an instant success -- combining high-flying traditional maneuvers with innovative offensive attacks of his own creation In less than a year Tajiri had moved on to the resurrected NWA to challenge (and lose to) Dan Severn for the Heavyweight Title Tajiri then moved on to Mexico under a mask (calling himself Aquarius) where he earned the coveted EMLL Light Heavyweight championship Tajiri later went back to his home country of Japan where his feuds with Shinjiro Otani and Jushin Liger wowed their stunned audiences By the time Tajiri had moved to ECW in 1998 to feud with Super Crazy Yoshihiro would earn himself several television titles; honing his skills for a move to the WWF in 2001.Which is exactly where he went that May Utilized in a short feud with Tazz and X-Pac Tajiri became the WWF Light Heavyweight Champion and even hooked up with Torrie Wilson for a short amount of time he's lost and won the Cruiserweight strap on several occasions grabbed the Tag Team belts with Eddie Guerrero and his currently feuding with masked star Rey Mysterio Tajiri just wont another Cruiserweight belt just a few weeks ago defeating Rey Mysterio in a hard-fought Smackdown melee Look for more good things to come from Tajiri in subsequent years Snapmare & Dropkick 1 - Circle + right It Thrusts Down - Circle (opponent near edge Raise the Opponent Up - Circle (opponent must be slumped down at lower turnbuckle) Turnbuckle Dropkick 2 - X (opponent must be slumped down at lower turnbuckle) Irish Whip - Circle (opponent must be leaning on ropes) Rolling Cradle Pin 1 - Circle (opponent must be dazed) Double Team Moves (against standing adversary) Dropkick & Rolling Clutch - Circle + down Back to Superstars... his new place isn't too far from Mom and Dad and you'll still get plenty of chances to see him Animal Adventure Park's youngest giraffe built by construction crews at the park in a month's time to ensure Tajiri has space to grow through the winter months Fans will still be able to catch up with him and check his progress because the new space is equipped to stream video from Tajiri's own webcam in Harpursville.  what will you see when you tune into Tajiri's livestream inside the barn In addition to the typical structure of a giraffe stall much like the space Tajiri shared with his mom which he currently shares with Bongo antelopes in separate stalls is slightly smaller than April and Oliver's the giraffe-designated space is bigger by about 100 square feet There's an automatic watering system — essentially water fountains — higher-quality insulation and heating sources Park owner Jordan Patch says the chute is a space keepers will train Tajiri to walk into in order to keep him stationary while he's given veterinary care While the distance between Tajiri's new home and the barn where he was born isn't far — about 500 yards across the park — moving a giraffe so building the barn was a quick turnaround during a time when not much is usually happening at Animal Adventure Park "We now have a giant project we're undertaking when we'd normally be pulling back and hunkering down," Patch said, "so our day-to-days are a little more stressful." all of the groundwork for the barn was prepared By the time the crews had spent a month building the barn and outdoor spaces ► April the Giraffe's son, Tajiri, to stay at Animal Adventure Park ► Animal Adventure Park owner celebrates personal joys ► Animal Adventure's Year of the Giraffe 2017 a professional transporter drove up from the Catskill area with a large aluminum livestock trailer tall enough to fit a giraffe calf and low enough to the ground to make entrance and exit easy When Tajiri made the successful move into his new bachelor pad "He is happy. His parents are happy. The team is happy," Animal Adventure Park posted on Facebook "We're working against three timelines," Patch said while the barn was being built Tajiri is at the natural age when a male calf in the wild would be driven out of the herd but most male giraffes are nomadic bachelors who leave their mother and go off on their own or form a bachelor group with other young males "It's designed by nature to avoid inbreeding," Patch said that natural instinct is simulated by separating young giraffes from their mothers at the appropriate time Originally, a plan was in place to send Tajiri to a different facility Animal Adventure Park partnered with the Wildlife Conservation Center of Virginia which planned to oversee the exhibit design buildout and transport of Tajiri to its new facility "We wholeheartedly were behind the transition of Tajiri to another facility," Patch said "It's about genetic diversity and a sustainable captive population But near the end of October, when the move was planned, the park changed its decision, citing requirements and unsatisfied deadlines in the given time frame which may soon be occupied by a new giraffe on the property "Females would be a long-term plan," Patch said in the barn built years back when the first giraffe stepped foot on the Animal Adventure Park property in Harpursville April the giraffe is in the midst of her second pregnancy at the park Announced in July, April's calf is due in March. big changes are likely in April's development as she enters the last trimester of a giraffe's 15-month gestation period and Patch says the park plans to launch several innovative and educational features to give fans of April the giraffe more ways to interact and learn while they watch and wait for the next baby giraffe to be born at Animal Adventure Park.  More: Here's everything you need to know about April the giraffe's pregnancy note the new barn and its yard take up about a third of the space originally designated for the park's new Wilds of Asia exhibit guests' first stop when they enter the park will be Tajiri the giraffe "The only way to do this correctly is also the only option," Patch said After his birth captivated millions of animal lovers throughout the world, Tajiri will soon say goodbye to Animal Adventure Park.  In a Facebook post Tuesday morning Animal Adventure Park announced that Tajiri Animal Adventure is partnering with the Wildlife Conservation Center of Virginia. The center will oversee the exhibit design build out and transport of Tajiri to their new facility "This move will allow Tajiri to play a vital role in the conservation and propagation initiatives that align with the facilities mission statement," the Facebook post stated Tajiri will be joined by female companions to "continue the genetic pool of healthy giraffes in management programs." Tajiri's departure will be documented for fans "A Tajiri cam is in your future," the post promises.  April the Giraffe sprang to fame in February of 2017 when a live cam streamed her pregnancy with Tajiri to the world.  More: IT'S A BOY! 'Perfect delivery' for April the Giraffe at Animal Adventure Park 2017 to an audience of about about 1.2 million viewers on the live YouTube stream and around 800,000 viewers of the park's Facebook Live video He was named by one of the park's former giraffe keepers after a naming contest revealed the most voted option to be "Allysa's choice." it was planned that he would leave the park for safety purposes According to a previous report by the Press & Sun Bulletin / pressconnects.com Tajiri was expected to leave the park after April raised him naturally and he has weaned himself after six to 12 months But the park won't be without a baby for long On July 25, after months of questions and speculation, it was confirmed that April is pregnant with her fifth calf. She is due in the spring of 2019. This pregnancy will be live-streamed as well.  Keep an eye on these social media platforms for updates: Follow @MaggieGilroy on Twitter More: Animal Adventure owner pulls application for funding, says park used as 'political pawn' More: Here we go again: Animal Adventure Park confirms April the giraffe is pregnant More: No giraffe news yet, but Animal Adventure Park's about to get bigger On Valentine's Day, Animal Adventure Park announced male giraffe Tajiri has a new companion An as-yet-unnamed female giraffe made her online debut Thursday when the Harpursville park launched a live video feed from Tajiri's barn, a second facility constructed at the park in November when the young calf moved out of the barn he shared with parents April and Oliver at the park Tajiri's companion is a 14-year-old female reticulated giraffe who is 14 feet tall She arrived at the park in January from another facility Park owner Jordan Patch compared this new pairing to the park's resident parent giraffes and emphasized the park's commitment to furthering the species "Tajiri is now one step closer to starting his own family," Patch said "which truly means that Taj now has the opportunity to fulfill his role in the species management program Tajiri is at the natural age when a male calf in the wild would be driven out of the herd so the park built a new barn with three stalls to house more giraffes but most male giraffes are nomadic bachelors who leave their mother and go off on their own or form a bachelor group with other young males "designed by nature to avoid inbreeding," Patch said More: Animal park owner's baby faces fraught health journey which he previously shared with Bongo antelopes in separate stalls Since Tajiri's new companion arrived at the facility unnamed the park has launched a naming contest to let giraffe fans chime in with their suggestions To vote, visit namethegiraffe.com. Votes are $5 for every five votes and the proceeds will in part support giraffe conservation initiatives the park's annual Ava's Little Heroes fundraiser and Animal Adventure Park Inside the Barn: Animal Adventure Park moves Tajiri away from April the Giraffe, but not far There will be two rounds of voting; the first began Thursday and runs through Feb Tajiri's new companion's name will be announced March 1 and Tajiri's first sibling, is expected in March In the barn built years back when the first giraffe stepped foot on the Animal Adventure Park property in Harpursville Announced in July 2018, April's calf is due in March. In the last trimester of a giraffe's 15-month gestation period Patch says the park plans to launch several innovative and educational features to give fans of April the giraffe more ways to interact and learn while they watch and wait for the next baby giraffe to be born at Animal Adventure Park.  Follow Katie Sullivan Borrelli on Twitter @ByKatieSullivan. Support our journalism and become a digital subscriber today. Click here for our special offers. By Craig T. Kojima