Ultrasensitive Imaging of Ca2+ Dynamics in Pancreatic Acinar Cells of Yellow Cameleon-Nano Transgenic Mice
AbstractYellow 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
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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.
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. International Journal of Molecular Sciences. 2014; 15(11):19971-19986.Chicago/Turabian Style
Oshima, Yusuke; Imamura, Takeshi; Shintani, Atsuko; Kajiura-Kobayashi, Hiroko; Hibi, Terumasa; Nagai, Takeharu; Nonaka, Shigenori; Nemoto, Tomomi. 2014. "Ultrasensitive Imaging of Ca2+ Dynamics in Pancreatic Acinar Cells of Yellow Cameleon-Nano Transgenic Mice." Int. J. Mol. Sci. 15, no. 11: 19971-19986.