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FUNCTIONAL DYMAMISM OF GASTRIC MUCOSA BY MEANS OF HIGH-PRESSURE FREEZING METHOD

高压冷冻法增强胃粘膜的功能活力

基本信息

批准号：
18590190
负责人：
SUGANUMA Tatsuo
金额：
$ 2.46万
依托单位：
UNIVERSITY OF MIYAZAKI
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590190/
关键词：
HIGH-PRESSURE FREEZING GASTRIC MUCOSA ELECTRON MICROSCOPE GASTRIC-PIT PARIETAL CELL ISOLATED GASTRIC MUCOSA STIMULATION OF ACID SECRETION HISTOCHEMISTRY AUTOPHAGOSOME 粘液壁細胞ラット

项目摘要

To elucidate a functional transformation of gastric parietal cells, we have newly developed an isolated rat gastric mucosa model whose parietal cells exhibited a reverting process from the active to the resting state of acid secretion. Immunohistochemistry of proton pump demonstrated a progressive translocation of proton pump from the apical to the cytoplasmic region. At the ultrastructural level, the apical membranes were subsequently internalized into the cytoplasm forming unique penta-laminar membranes. They then formed a number of multilamellar autophagosomes that were intensely labeled for proton pump. The parietal cells exhibited well-developed Golgi apparatus and lysosomal compartments involving the multilamellar membranes. The present findings provide new insights into the functional transformation in gastric parietal cells reverting to their resting conformation.It is clinicopathologically important to elucidate the cell kinetics for the maintenance of normal gastric epithelium. In rat gastric mucosa isolated after stimulation, a number of cells were exfoliated into the gastric lumen of the pit region. It was undertaken to clarify the origin of exfoliated cells and their histochemical profiles by taking the advantages of cryotechniques. Most of the exfoliated cells were identified as pit-parietal cells labeled with both peanut-lectin and anti-proton pump antibody. Quantitative analysis verified a time-dependent increase in the number of exfoliated cells in the gastric mucosa isolated after stimulation. Most of the exfoliated cells were negative to the apoptotic markers such as TUNEL staining and caspase-3. Ultrastructurally, autophagusome-like structures consisting of proton pump positive membranes were frequently seen in the exfoliated pit-parietal cells. In addition, the pit-parietal cell exfoliation was accompanied by sealing of their basal portion with the cytoplasmic processes of adjacent surface mucous cells.

为了阐明胃壁细胞的功能转化，我们新建立了离体大鼠胃粘膜模型，其胃壁细胞从分泌酸的活跃状态恢复到分泌酸的静止状态。质子泵免疫组化显示质子泵从根尖区向细胞质区进行性移位。在超微结构水平上，顶膜随后内化到细胞质中，形成独特的五层膜。然后它们形成了许多多层自噬体，这些自噬体被强烈地标记为质子泵。壁细胞显示发育良好的高尔基体和溶酶体室，包括多层膜。本研究结果为胃壁细胞恢复到其静息构象的功能转化提供了新的见解。阐明细胞动力学对维持正常胃上皮具有重要的临床病理学意义。在刺激后分离的大鼠胃粘膜中，许多细胞脱落到胃坑区胃腔内。利用冷冻技术的优势，阐明脱落细胞的起源及其组织化学特征。大部分脱落细胞被鉴定为核壁细胞，同时被花生凝集素和抗质子泵抗体标记。定量分析证实了刺激后胃粘膜脱落细胞数量的时间依赖性增加。大部分脱落细胞TUNEL染色、caspase-3等凋亡标志物均为阴性。超微结构上，在脱落的窝壁细胞中可见由质子泵正膜组成的自噬体样结构。此外，凹壁细胞的脱落伴随着其基部与邻近表面黏液细胞的细胞质突起的封闭。