Metabolic and functional analysis of pancreatic islet cells using the fluorescent ATP probe gene knock-in mouse

• Project/Area Number: 16K09746
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Metabolomics
• Research Institution: Kyoto University
• Principal Investigator: NAGASHIMA KAZUAKI 京都大学, 医学研究科, 客員研究員 (40324628)
• Project Period (FY): 2016-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2019: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2017: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2016: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
• Keywords: 膵β細胞 / ATP / 不均一性 / 糖代謝異常 / エネルギー・糖代謝異常 / イオンチャネル / 膵島細胞

Outline of Final Research Achievements

Using pancreatic islet cells from fluorescent ATP probe gene knock-in mouse and the fluorescent Ca2+ probe Fura-2AM, we have carried out dual measurement of intracellular ATP([ATP]c) and Ca2+ ([Ca]c) concentrations and found that even beta cells in the same islet have different response to the stimuli. Using single-cell transcriptome analysis, we also found heterogeneity among beta cells in gene expressions. These data suggest beta cells in an islet are heterogeneous both functionally and genetically.

Academic Significance and Societal Importance of the Research Achievements

ATPは細胞内代謝に深く関わる。血糖低下させる唯一のホルモンであるインスリンは膵β細胞から分泌され、その機序は代謝説で説明されているが未だ詳細不詳な点が多い。本研究では、細胞内ATP濃度を測定可能な蛍光ATPプローブノックインマウス膵島とCa2+インジケーターFura2を用いて細胞内代謝変化を代謝・機能的に検討、個々のβ細胞遺伝子発現も検討し、同じβ細胞間でも代謝・機能的にも遺伝子発現的にも不均一であること、膵島内では様々な状態の膵β細胞が共存して存在（代謝・機能的にも遺伝子発現的にも不均一なβ細胞が共存）している可能性が示唆された。

Research Products

• [Int'l Joint Research] University of British Columbia(カナダ)
• [Int'l Joint Research] University of Geneva(スイス)
• [Int'l Joint Research] University of British Columbia(カナダ)
• [Int'l Joint Research] University of Geneva(スイス)
• [Int'l Joint Research] University of British Columbia(Canada)
• [Int'l Joint Research] University of Geneva(Switzerland)
• [Journal Article] Transcriptome Analysis Dissects the Replicating Process of Pancreatic Beta Cells in Partial Pancreatectomy Model (2020)
  • Author(s): Tatsuoka H, Sakamoto S, Yabe D, Kabai R, Kato U, Okumura T, Botagarova A, Tokumoto S, Usui R, Ogura M, Nagashima K, Mukai E, Fujtani Y, Watanabe A, Inagaki N
  • Journal Title: iScience Volume: 23 Issue: 12 Pages: 101774-101774
  • DOI: 10.1016/j.isci.2020.101774
  • Peer Reviewed / Open Access
• [Journal Article] GPR40 activation initiates store-operated Ca2+ entry and potentiates insulin secretion via the IP3R1/STIM1/Orai1 pathway in pancreatic β-cells (2019)
  • Author(s): Usui R, Yabe D, Fauzi M, Goto H, Botagarova A, Tokumoto S, Tatsuoka H, Tahara Y, Kobayashi S, Manebe T, Baba Y, Kurosaki T, Herrera PL, Ogura M, Nagashima K, Inagaki N
  • Journal Title: Sci Rep. Volume: 9 Issue: 1 Pages: 15562-15562
  • DOI: 10.1038/s41598-019-52048-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Store-operated Ca2+ entry activated by STIM1 plays an essential role in GPR40-mediated GIIS potentiation (2019)
  • Author(s): Usui R, Yabe D, Fauzi M, Goto H, Botagarova A, Tokumoto S, Tatsuoka H, Tahara Y, Kobayashi S, Manabe T, Baba Y, Kurosaki T, Ogura M, Nagashima K, Inagaki N
  • Organizer: American Diabetes Association the 79th Scientific Sessions
  • Int'l Joint Research
• [Presentation] ER Ca2+ sensor, stromal interaction molecule 1 (STIM1) plays an important role in GPR40-mediated potentiation of glucose-induced insulin secretion. (2018)
  • Author(s): Usui R, Yabe D, Goto H, Fauzi M, Botagarova A, Tokumoto S, Tatsuoka H, Tahara Y, Ogura M, Nagashima K, Inagaki N
  • Organizer: Asian Islet Biology and Incretin 2018
  • Int'l Joint Research
• [Presentation] The role of ER Ca2+sensor, stromal interaction molecule 1 (STIM1) in GPR40-mediated potentiation of glucose-induced insulin secretion. (2018)
  • Author(s): Usui R, Yabe D, Goto H, Fauzi M, Tokumoto S, Tatsuoka H, Tahara Y, Ogura M, Nagashima K, Inagaki N
  • Organizer: 54th Annual Meeting of the European Association for the Study of Diabetes
  • Int'l Joint Research
• [Presentation] ATP and Ca2+ dynamics in pancreatic β-cells:Ex vivo analysis using ATP biosensor GO-ATeam1 (2017)
  • Author(s): Usui R, Yabe D, Fauzi M,Tokumoto S, Tatsuoka H, Tahara Y, Ogura M, Nagashima K, Inagaki N.
  • Organizer: The 9th Scientific Meeting of the Asian Association for the Study of Diabetes (AASD)
  • Int'l Joint Research
• [Presentation] Genetic and epigenetic programs for regenerating pancreatic beta-cells following partial pancreatectomy in mice (2017)
  • Author(s): Tatsuoka H, Nakamura M, Yabe D, Usui R, Muhammad F, Tokumoto S, Tahara Y, Ogura M, Nagashima K, Watanabe A, Inagaki N
  • Organizer: Kyoto Diabetes Mini-Symposium: Beta Cell Replacement Strategies.
  • Int'l Joint Research
• [Presentation] 蛍光ATPプローブ遺伝子ノックインマウスを用いた膵β細胞内代謝解析 (2016)
  • Author(s): 杉崎 和, 山本正道, 龍岡久登, 臼井亮太, 今村博臣, 田原裕美子, 小倉かさね, 佐藤広規, 佐藤雄一, 小倉雅仁, 長嶋一昭, 稲垣暢也.
  • Organizer: 第59回日本糖尿病学会年次学術集会
  • Place of Presentation: 京都
  • Year and Date: 2016-05-19