Ultrasensitive Imaging of Ca2+ Dynamics in Pancreatic Acinar Cells of Yellow Cameleon-Nano Transgenic Mice

International Journal of Molecular Sciences

Atsuko Shintani¹  Shigenori Nonaka¹  Hiroko Kajiura-Kobayashi¹  Tomomi Nemoto²  Terumasa Hibi²  Yusuke Oshima³  Takeshi Imamura³  Takeharu Nagai⁴ 

[1] Laboratory for Spatiotemporal Regulations, National Institute for Basic Biology, Okazaki,Aichi 444-8585, Japan;Laboratory of Molecular and Cellular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan;Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon City, Ehime 791-0295, Japan;The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567-0047, Japan;

关键词: Yellow Cameleon;    Ca2+ indicator;    Ca2+ imaging;    Förster resonance energy transfer (FRET);    genetically encoded Ca2+ indicators (GECI);    Yellow Cameleon-Nano(YC-Nano);    two-photon excitation fluorescence microscopy;    live-cell imaging;    exocytotsis;    acinar cell;   

DOI  :  10.3390/ijms151119971

来源: DOAJ

【 摘 要 】

Yellow Cameleons are genetically encoded Ca2+ indicators in which cyan and yellow fluorescent proteins and calmodulin work together as a fluorescence (Förster) resonance energy transfer Ca2+-sensor probe. To achieve ultrasensitive Ca2+ imaging for low resting Ca2+ or small Ca2+ transients in various organs, we generated a transgenic mouse line expressing the highest-sensitive genetically encoded Ca2+ indicator (Yellow Cameleon-Nano 15) in the whole body. We then focused on the mechanism of exocytotic events mediated by intracellular Ca2+ signaling in acinar cells of the mice with an agonist and observed themby two-photon excitation microscopy. In the results, two-photon excitation imaging of Yellow Cameleon-Nano 15 successfully visualized intracellular Ca2+ concentrationunder stimulation with the agonist at nanomolar levels. This is the first demonstrationfor application of genetically encoded Ca2+ indicators to pancreatic acinar cells. We also simultaneously observed exocytotic events and an intracellular Ca2+ concentration under in vivo condition. Yellow Cameleon-Nano 15 mice are healthy and no significant deteriorative effect was observed on physiological response regarding the pancreatic acinar cells. The dynamic range of 165% was calculated from Rmax and Rmin values under in vivo condition. The mice will be useful for ultrasensitive Ca2+ imaging in vivo.

【 授权许可】

Unknown