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活体双光子成像在黄色 Cameleon 转基因小鼠分泌器官中的钙信号研究。

Intravital Two-photon Imaging of Ca signaling in Secretory Organs of Yellow Cameleon Transgenic Mice.

机构信息

Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan.

Department of Immunology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.

出版信息

Sci Rep. 2018 Oct 26;8(1):15880. doi: 10.1038/s41598-018-34347-1.

Abstract

Intracellular calcium ([Ca]i) signaling regulates physiological functions in most cells. In secretory organs, such as the pancreas, salivary gland, and lacrimal gland (LG), [Ca]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 [Ca]i changes using mouse line expressing Yellow Cameleon 3.60, a genetically encoded Ca indicator. Elevation of [Ca]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 [Ca]i response to cholinergic stimulation was induced under pathological conditions by postganglionic denervation. Intravital Ca imaging in secretory organs will broaden our understanding of the cellular mechanisms in animal models of secretory diseases.

摘要

细胞内钙 ([Ca]i) 信号调节大多数细胞的生理功能。在胰腺、唾液腺和泪腺 (LG) 等分泌器官中,腺泡细胞中的 [Ca]i 升高会引发液体分泌,这在整个生命周期中对维持器官功能健康起着至关重要的作用。了解分泌器官的分泌机制很重要,但由于缺乏适用于活体动物的分析系统,限制了研究范围,难以深入了解分泌的精确机制。我们建立了一种用于分泌器官特定细胞类型的活体成像系统,使用表达 Yellow Cameleon 3.60 的小鼠系来监测 [Ca]i 变化,Yellow Cameleon 3.60 是一种基因编码的 Ca 指示剂。在离体和活体条件下,我们可以监测到特定细胞类型的分泌器官中 [Ca]i 的升高。我们发现,在神经节后去神经支配的病理条件下,LG 对胆碱能刺激的 [Ca]i 反应明显减弱。活体分泌器官的 Ca 成像将拓宽我们对分泌性疾病动物模型中细胞机制的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f599/6203801/fc4ee91d58c5/41598_2018_34347_Fig1_HTML.jpg

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