Genetic Ablation of the ClC-2 Cl- Channel Disrupts Mouse Gastric Parietal Cell Acid Secretion.
Genetic Ablation of the ClC-2 Cl- Channel Disrupts Mouse Gastric Parietal Cell Acid Secretion.
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DOI：
10.1371/journal.pone.0138174
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Blikslager AT
中科院分区：
文献类型：
--
作者：
Nighot MP;Nighot PK;Ma TY;Malinowska DH;Shull GE;Cuppoletti J;Blikslager AT
本研究旨在使用WT和ClC-2-/-小鼠胃检查ClC-2消融对细胞形态、壁细胞丰度、H/K ATP酶表达、壁细胞超微结构和酸分泌的影响。细胞的组织学，形态学和蛋白质进行了检查，使用成像技术，电子显微镜和蛋白质印迹。测定组胺对胃内容物pH值的影响。还使用先前建立的方法和促分泌素测量酸分泌，以获得最大的酸分泌和形态变化。与WT相比，ClC-2-/-胃粘膜组织学组织似乎被破坏，包括胃腺扩张、胃腺区域缩短和所有细胞层的解体。壁细胞数和H/K ATP酶表达分别显著减少34%（P<0.05）和53%（P<0.001），电镜观察胞质小管囊泡明显减少，但小管未扩张。在WT壁细胞，ClC-2是明显的，在一个类似的细胞位置的H/K ATP酶的免疫荧光，并出现与tubulovesicles的免疫金电子显微镜。与WT相比，在ClC-2-/-小鼠中胃内容物的组胺刺激的[H+]显著（P<0.025）降低9.4倍（89%）。与WT相比，在ClC-2-/-中组胺/卡巴胆碱刺激的胃酸分泌显著减少（范围84- 95%，P<0.005），而胃蛋白酶原分泌不受影响。ClC-2的基因切除导致胃腺区域减少，壁细胞数量减少，H/K ATP酶减少，小管囊泡减少和刺激酸分泌减少。
The present studies were designed to examine the effects of ClC-2 ablation on cellular morphology, parietal cell abundance, H/K ATPase expression, parietal cell ultrastructure and acid secretion using WT and ClC-2-/- mouse stomachs. Cellular histology, morphology and proteins were examined using imaging techniques, electron microscopy and western blot. The effect of histamine on the pH of gastric contents was measured. Acid secretion was also measured using methods and secretagogues previously established to give maximal acid secretion and morphological change. Compared to WT, ClC-2-/- gastric mucosal histological organization appeared disrupted, including dilation of gastric glands, shortening of the gastric gland region and disorganization of all cell layers. Parietal cell numbers and H/K ATPase expression were significantly reduced by 34% (P<0.05) and 53% (P<0.001) respectively and cytoplasmic tubulovesicles appeared markedly reduced on electron microscopic evaluation without evidence of canalicular expansion. In WT parietal cells, ClC-2 was apparent in a similar cellular location as the H/K ATPase by immunofluorescence and appeared associated with tubulovesicles by immunogold electron microscopy. Histamine-stimulated [H+] of the gastric contents was significantly (P<0.025) lower by 9.4 fold (89%) in the ClC-2-/- mouse compared to WT. Histamine/carbachol stimulated gastric acid secretion was significantly reduced (range 84–95%, P<0.005) in ClC-2-/- compared to WT, while pepsinogen secretion was unaffected. Genetic ablation of ClC-2 resulted in reduced gastric gland region, reduced parietal cell number, reduced H/K ATPase, reduced tubulovesicles and reduced stimulated acid secretion.