Intravital Two-photon Imaging of Ca2+ signaling in Secretory Organs of Yellow Cameleon Transgenic Mice

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作者
Kai Jin
Toshihiro Imada
Shigeru Nakamura
Yusuke Izuta
Erina Oonishi
Michiko Shibuya
Hisayo Sakaguchi
Takahiro Adachi
Kazuo Tsubota
机构
[1] Keio University School of Medicine,Department of Ophthalmology
[2] Tokyo Medical and Dental University,Department of Immunology, Medical Research Institute
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关键词
Secretory Organs; Yellow Cameleon (YC); Intravital Imaging; Pancreatic Acinar Cells; FRET Ratio;
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摘要
Intracellular calcium ([Ca2+]i) signaling regulates physiological functions in most cells. In secretory organs, such as the pancreas, salivary gland, and lacrimal gland (LG), [Ca2+]i elevation in acinar cells triggers fluid secretion, which plays vital roles in the maintenance of functional health across the life-course. It is important to understand the secretory mechanism of secretory organs, but lack of analytic systems available for living animals limits the scope of research to gain deeper insights into the precise mechanism of secretion. We established an intravital imaging system for specific cell types of secretory organs to monitor the [Ca2+]i changes using mouse line expressing Yellow Cameleon 3.60, a genetically encoded Ca2+ indicator. Elevation of [Ca2+]i in specific cell types of secretory organs could be monitored after cholinergic stimulation ex vivo and intravitally. We found that a marked attenuation of LG [Ca2+]i response to cholinergic stimulation was induced under pathological conditions by postganglionic denervation. Intravital Ca2+ imaging in secretory organs will broaden our understanding of the cellular mechanisms in animal models of secretory diseases.
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