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Immunohistochemical detection of cell components related to the acid secreting dynamics in gastric parietal cells.

免疫组织化学检测与胃壁细胞酸分泌动力学相关的细胞成分。

基本信息

批准号：
15590164
负责人：
TSUYAMA Shinichiro
金额：
$ 2.3万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590164/
关键词：
Stomach Parietal cell Proton pump Distribution in gland Ezrin Chloride channel High pressure freezing Freeze substitution プトトンポンプ胃底腺胃塩酸分泌休止 H^+K^+-ATPase ガストリンオメプラゾール急速凍結

项目摘要

The main purpose of this study was to ascertain whether or not the cells are functionally similar throughout the gland, and how transformation mechanism with cytoskeletal components is related to the distribution of ezrin molecules and chloride channel. In this study, the specimens were processed with cryofixation method showed good morphological preservation and retention of soluble components at the light and electron microscopic level. To suppress severe ice-crystal damage or loss of their immunocytochemical properties, stomach tissues from ad libitum-fed or starved rats were prepared with rapid freezing followed by freeze-substitution for light microscopy, and with high pressure freezing for electron microscopy. The transformable structure of parietal cell was examined between the active and resting phases of acid secretion. These structures were the dilated intracellular canaliculi, including numerous microvilli in the secreting phase, and the developed tubulovesicles in apical cy … More toplasm at resting phase. The immunohistochemical staining with anti-alpha- or beta-subunits of H^+・K^+-ATPase antibodies was localized on the cells from both phases. Labeling was more intense and clear on the membranes of intracellular canaliculi, microvilli and tubulovesicles than that with chemical fixation. The positively stained parietal cells were scattered widely throughout the gland. The cells in the secreting phase located in the upper one third of the gland were densely stained with an anti-phosphorylated ezrin antibody. In the resting phase, the antibody showed a weak reaction. However, the anti-nonphosphorylated ezrin antibody hardly labeled those cells. Anti-chloride channel antibody immunostaining showed a narrow zone of positive reaction around the neck region in the secreting phase. These labelings were observed on the membrane of intracellular canaliculi, microvilli and tubulovesicles. It was concluded that the cells in around the upper one third are more active cells for functional secretion than those of other segments, while parietal cells were distributed throughout the gastric gland. Less

本研究的主要目的是确定整个腺体中的细胞功能是否相似，以及细胞骨架成分的转化机制如何与ezrin分子和氯离子通道的分布相关。本研究中，冷冻固定法处理的标本在光镜和电镜水平上表现出良好的形态保存和可溶性成分的保留。为了抑制严重的冰晶损伤或其免疫细胞化学性质的损失，从自由采食或饥饿大鼠胃组织制备快速冷冻，然后冷冻替代光学显微镜，并与高压冷冻电镜。观察了壁细胞泌酸活动期与静息期之间的结构变化。这些结构是：分泌期细胞内小管扩张，包括大量微绒毛，顶端囊泡发达， ...更多信息休眠期的细胞质。用H^+·K^+-ATP酶α亚基或β亚基抗体对两个时期的细胞进行免疫组织化学染色。细胞内小管、微绒毛和管泡膜上的标记比化学固定法更强、更清晰。阳性染色的壁细胞广泛分布于整个腺体。位于腺体上三分之一的分泌期细胞用抗磷酸化埃兹蛋白抗体密集染色。在静止期，抗体显示弱反应。然而，抗非磷酸化埃兹蛋白抗体几乎没有标记这些细胞。抗氯离子通道抗体免疫组化染色显示分泌期颈部周围有一狭窄的阳性反应区。这些标记在细胞内小管、微绒毛和管泡的膜上观察到。结果表明，胃上1/3部的细胞是功能性分泌较活跃的细胞，而壁细胞则分布于整个胃腺。少