Localization of ClC-2 Cl- channels in rabbit gastric mucosa.
Localization of ClC-2 Cl- channels in rabbit gastric mucosa.
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DOI：
10.1152/ajpcell.2001.280.6.c1599
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发表时间：
2001-06
期刊：
影响因子：
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通讯作者：
A. M. Sherry;D. Malinowska;R. Morris;G. Ciraolo;J. Cuppoletti
中科院分区：
文献类型：
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作者：
A. M. Sherry;D. Malinowska;R. Morris;G. Ciraolo;J. Cuppoletti
穿过壁细胞顶分泌膜的 HCl 分泌涉及 H+-K+-ATP 酶、ClC-2 Cl- 通道和 K+ 通道。在本研究中，确定了兔胃粘膜和离体胃腺中 ClC-2 mRNA 和蛋白质的细胞和亚细胞分布。通过原位杂交和直接原位 RT-PCR 将 ClC-2 mRNA 定位于壁细胞。通过免疫过氧化物酶显微镜检查，ClC-2蛋白集中在壁细胞中。免疫荧光共聚焦显微镜表明，ClC-2 定位于受刺激壁细胞的分泌小管膜和静息壁细胞的细胞内结构。免疫金电子显微镜证实，ClC-2 存在于受刺激细胞的分泌小管膜和静息壁细胞的管小泡中。这些发现，连同先前对天然和重组通道的功能表征，强烈表明 ClC-2 是 Cl- 通道，它与 H+-K+-ATP 酶和 K+ 通道一起，导致 HCl 分泌穿过壁细胞分泌膜。
HCl secretion across the parietal cell apical secretory membrane involves the H+-K+-ATPase, the ClC-2 Cl- channel, and a K+ channel. In the present study, the cellular and subcellular distribution of ClC-2 mRNA and protein was determined in the rabbit gastric mucosa and in isolated gastric glands. ClC-2 mRNA was localized to parietal cells by in situ hybridization and by direct in situ RT-PCR. By immunoperoxidase microscopy, ClC-2 protein was concentrated in parietal cells. Immunofluorescent confocal microscopy suggested that the ClC-2 was localized to the secretory canalicular membrane of stimulated parietal cells and to intracellular structures of resting parietal cells. Immunogold electron microscopy confirmed that ClC-2 is in the secretory canalicular membrane of stimulated cells and in tubulovesicles of resting parietal cells. These findings, together with previous functional characterization of the native and recombinant channel, strongly indicate that ClC-2 is the Cl- channel, which together with the H+-K+-ATPase and a K+ channel, results in HCl secretion across the parietal cell secretory membrane.