Visualizing calcium responses to acetylcholine convection along endothelium of arteriolar networks in Cx40BAC-GCaMP2 transgenic mice
Visualizing calcium responses to acetylcholine convection along endothelium of arteriolar networks in Cx40BAC-GCaMP2 transgenic mice
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DOI：
10.1152/ajpheart.00425.2011
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发表时间：
2011-09-01
影响因子：
4.8
通讯作者：
Segal, Steven S.
中科院分区：
文献类型：
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作者：
Bagher, Pooneh;Davis, Michael J.;Segal, Steven S.
Bagher P，Davis MJ，Segal SS.在Cx40（BAC）-GCaMP 2转基因小鼠中，钙对乙酰胆碱沿小动脉网络内皮沿着对流的响应可视化。美国生理学杂志心脏循环生理学301：H794-H802，2011年。首次发表于2011年6月10日; doi：10.1152/ajpheart.00425.2011。乙酰胆碱引起内皮依赖性血管舒张，随后细胞内钙升高。尽管在人类和动物研究中广泛应用，但在体内对血管内ACh的钙反应尚未可视化。微离子电渗法的乙酰胆碱在组织邻近的小动脉激活的“局部”扩散区域内的内皮细胞上的腔外毒蕈碱受体，但它是未知的，是否乙酰胆碱释放在这种方式获得访问的流动流导致进一步的行动下游。为了验证这一假设并提供体内钙信号转导的新见解，我们研究了在小动脉内皮细胞中选择性表达荧光钙传感器GCaMP 2的麻醉雄性Cx40 BAC-GCaMP 2转基因小鼠（n = 22; 5-9 mo; 33 +/- 1 g）的提睾肌微循环。次最大ACh刺激使用微离子导入法（1 μ m移液管尖端，500 nA）。刺激持续时间
Bagher P, Davis MJ, Segal SS. Visualizing calcium responses to acetylcholine convection along endothelium of arteriolar networks in Cx40(BAC)-GCaMP2 transgenic mice. Am J Physiol Heart Circ Physiol 301: H794-H802, 2011. First published June 10, 2011; doi:10.1152/ajpheart.00425.2011.-Acetylcholine evokes endothelium-dependent vasodilation subsequent to a rise in intracellular calcium. Despite widespread application in human and animal studies, calcium responses to intravascular ACh have not been visualized in vivo. Microiontophoresis of ACh in tissue adjacent to an arteriole activates abluminal muscarinic receptors on endothelial cells within a "local" region of diffusion, but it is unknown whether ACh released in such fashion gains access to the flow stream resulting in further actions downstream. To test this hypothesis and provide new insight into calcium signaling in vivo, we studied the cremaster muscle microcirculation of anesthetized male Cx40BAC-GCaMP2 transgenic mice (n = 22; 5-9 mo; 33 +/- 1 g) expressing the fluorescent calcium sensor GCaMP2 selectively in arteriolar endothelial cells. Submaximal ACh stimuli were delivered using microiontophoresis (1-mu m pipette tip, 500 nA). With stimulus duration