Molecular dissection of HCl secretion in gastric parietal cells using streptolysin O permeabilization.
Molecular dissection of HCl secretion in gastric parietal cells using streptolysin O permeabilization.
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DOI：
10.1007/978-1-59745-178-9_16
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发表时间：
2008
影响因子：
--
通讯作者：
Xia Ding;Fang Wu;Zhen Guo;X. Yao
中科院分区：
文献类型：
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作者：
Xia Ding;Fang Wu;Zhen Guo;X. Yao
组胺刺激的胃酸分泌涉及细胞内Ca2+的短暂升高和环腺苷单磷酸（cAMP）依赖性蛋白激酶a （PKA）通过磷酸化级联，其作用最终导致含有H， k - atp酶（腺苷三磷酸酶）的囊泡融合到顶壁细胞的顶质膜。为了剖析胃酸分泌背后的信号事件，我们利用溶链素O （SLO）建立了一个渗透胃腺模型。该模型的优点是它能够保留分泌机制所需的细胞质成分，同时允许大分子进入细胞质。我们的研究表明，慢速渗透腺体的酸分泌是一个camp依赖的过程，通过生化分析判断，这涉及到富含H， k - atp酶的管状囊泡向根尖质膜的募集。这些研究建立了一种功能渗透腺模型，其中静息到分泌的转变可以由第二信使触发，而细胞质环境的操纵可以很容易地实现。
Histamine-stimulated gastric acid secretion involves a transient elevation of intracellular Ca2+and the cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) cascade through phosphorylation, the actions of which ultimately result in the fusion of vesicles containing H,K-ATPase (adenosine triphosphatase) to the apical plasma membrane of parietal cells. To dissect the signaling events underlying gastric acid secretion, we have developed a permeabilized gastric gland model using streptolysin O (SLO). The advantage of this model is its ability to retain cytosolic components that are required for the secretory machinery while granting accessibility for the introduction of macromolecules into the cytoplasm. Our studies showed that acid secretion in SLO-permeabilized glands is a cAMP-dependent process and involves the recruitment of H,K-ATPase-rich tubulovesicles into the apical plasma membrane as judged by biochemical assays. These studies established a functional permeabilized gland model in which the resting-to-secreting transition can be triggered by second messengers, while the manipulation of the cytoplasmic environment can be achieved with ease.