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Open AccessArticle

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

1
Molecular Medicine for Pathogenesis, Graduate School of Medicine, Ehime University, Toon City, Ehime 791-0295, Japan
2
Division of Bio-Imaging, Proteo-Science Center, Ehime University, Toon City, Ehime 791-0295, Japan
3
Translational Research Center, Ehime University Hospital, Toon City, Ehime 791-0295, Japan
4
Laboratory for Spatiotemporal Regulations, National Institute for Basic Biology, Okazaki, Aichi 444-8585, Japan
5
Laboratory of Molecular and Cellular Biophysics, Research Institute for Electronic Science, Hokkaido University, Sapporo, Hokkaido 001-0020, Japan
6
The Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Osaka 567-0047, Japan
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2014, 15(11), 19971-19986; https://doi.org/10.3390/ijms151119971
Received: 9 September 2014 / Revised: 22 October 2014 / Accepted: 28 October 2014 / Published: 3 November 2014
(This article belongs to the Special Issue Laser Application in Life Sciences)
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 Rmax and Rmin values under in vivo condition. The mice will be useful for ultrasensitive Ca2+ imaging in vivo. View Full-Text
Keywords: 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 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
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Oshima, Y.; Imamura, T.; Shintani, A.; Kajiura-Kobayashi, H.; Hibi, T.; Nagai, T.; Nonaka, S.; Nemoto, T. Ultrasensitive Imaging of Ca2+ Dynamics in Pancreatic Acinar Cells of Yellow Cameleon-Nano Transgenic Mice. Int. J. Mol. Sci. 2014, 15, 19971-19986.

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