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

International Journal of Molecular Sciences

Yusuke Oshima³  Takeshi Imamura³  Atsuko Shintani⁴  Hiroko Kajiura-Kobayashi⁴  Terumasa Hibi¹  Takeharu Nagai²  Shigenori Nonaka⁴  Tomomi Nemoto¹ 

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

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: mdpi

【 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 them by two-photon excitation microscopy. In the results, two-photon excitation imaging of Yellow Cameleon-Nano 15 successfully visualized intracellular Ca²⁺ concentration under stimulation with the agonist at nanomolar levels. This is the first demonstration for 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 R_max and R_min values under in vivo condition. The mice will be useful for ultrasensitive Ca²⁺ imaging in vivo.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.