期刊
JOURNAL OF NEUROSCIENCE
卷 34, 期 47, 页码 15638-15647出版社
SOC NEUROSCIENCE
DOI: 10.1523/JNEUROSCI.5290-13.2014
关键词
cAMP; capacitance measurements; exocytosis; HCN channels; lactotrophs; RT-PCR
资金
- Ministry of Higher Education, Sciences, and Technology of the Republic of Slovenia [P3 310, J3 3654, J3 4051, J3 4146]
- Portuguese Foundation of Science and Technology of the Portuguese Ministry of Sciences, Technology, and High Education [441.00 Slovenia, SFRH/BD/41217/2007]
Hormone and neurotransmitter release from vesicles is mediated by regulated exocytosis, where an aqueous channel-like structure, termed a fusion pore, is formed. It was recently shown that second messenger cAMP modulates the fusion pore, but the detailed mechanisms remain elusive. In this study, we asked whether the hyperpolarization-activated cyclic nucleotide-gated (HCN) channels, which are activated by cAMP, are involved in the regulation of unitary exocytic events. By using the Western blot technique, a real-time PCR, immunocytochemistry in combination with confocal microscopy, and voltage-clamp measurements of hyperpolarizing currents, we show that HCN channels are present in the plasma membrane and in the membrane of secretory vesicles of isolated rat lactotrophs. Single vesicle membrane capacitance measurements of lactotrophs, where HCN channels were either augmented by transfection or blocked with an HCN channel blocker (ZD7288), show modulated fusion pore properties. We suggest that the changes in local cation concentration, mediated through HCN channels, which are located on or near secretory vesicles, have an important role in modulating exocytosis.
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