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Date:
2021-02-05
[Abstract] Connexins are membrane bound proteins that facilitate direct and local paracrine mediated cell-to-cell communication through their ability to oligomerise into hexameric hemichannels. When neighbouring channels align, they form gap-junctions that provide a direct route for information transfer between cells. In contrast to intact gap junctions, which typically open under physiological conditions, undocked hemichannels have a low open probability and mainly open in response to injury. Hemichannels permit the release of small molecules and ions (approximately 1kDa) into the local intercellular environment, and excessive expression/activity has been linked to a number of disease conditions. Carboxyfluorescein dye uptake measures functional expression of hemichannels, where increased ...
[摘要] [摘要]连接蛋白是有助于直接和局部旁分泌通过自己的能力介导的细胞-细胞通信膜结合蛋白寡聚成六聚半通道。当相邻通道对齐时,它们形成间隙连接,为单元之间的信息传输提供直接路径。与通常在生理条件下打开的完整的间隙连接相反,未对接的半通道的打开概率较低,主要是在受伤时打开。半通道允许小分子和离子(约1kDa)释放到局部细胞间环境中,并且过度表达/活性 与许多疾病状况有关。羧基荧光素染料的摄取量度了半通道的功能性表达,其中增加的半通道活性/功能反映了增加的负荷。该技术依赖于通过开放性半通道对不可渗透膜的荧光示踪剂的吸收,可用于比较在不同条件下(例如对照与疾病)培养的细胞单层之间的通道活性。其他技术(例如生物素化和电生理学)可以分别测量细胞表面表达和半通道开放可能性,但是,羧基荧光素的摄取提供了一种简单,快速且经济高效的方法来测定多种细胞类型的体外半通道活性。
图形摘要:
使用染料吸收量度半通道活性
[背景]连接蛋白(CX )是一体的跨膜蛋白,其寡聚到连接子在细胞表面。连接子与相似的六聚体蛋白复合物停靠在相邻细胞上,形成用于间隙连接细胞间通讯的双向导管(GJIC;Bosco等人,2011 ...
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