Summary
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.
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Acknowledgments
We would like to thank Professor John G. Forte for insightful discussion and support. This work was supported by grants from the Chinese Natural Science Foundation, Chinese 973 project (2002CB713700), Chinese 863 project (2001AA215331), and the National Institutes of Health (DK-56292) to X.Y. X.Y. is a GCC Distinguished Cancer Research Scholar.
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© 2008 Humana Press, a part of Springer Science+Business Media, LLC
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Ding, X., Wu, F., Guo, Z., Yao, X. (2008). Molecular Dissection of HCl Secretion in Gastric Parietal Cells Using Streptolysin O Permeabilization. In: Ivanov, A.I. (eds) Exocytosis and Endocytosis. Methods in Molecular Biology, vol 440. Humana Press. https://doi.org/10.1007/978-1-59745-178-9_16
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DOI: https://doi.org/10.1007/978-1-59745-178-9_16
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