Dynamics of gastric mucous gel layer in vivo and in vitro using high pressure freezing method

高压冷冻法体内外胃粘膜凝胶层动力学研究

• 批准号: 16590148
• 负责人: SUGANUMA Tatsuo
• 金额: $ 2.18万
• 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: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590148/
• 关键词: high pressure freezing; gastric gland; electron microscopy; histochemistry; gastric acid secretion; isolated gastric mucosa; parietal cell; mucin; 凍結置換; 胃; 粘液細胞

Cryofixation is currently accepted as the best initial fixation step to preserve not only the fine structure but also the antigenicity of biological specimens. To elucidate the functional transformation of gastric parietal cells, we have newly developed an in vitro experimental model, named the isolated gastric mucosa. In this study, acid secretion of the parietal cell was stimulated with histamine or inhibited with cimetidine, and the samples were cryofixed by plunge freezing for light microscopy or high-pressure freezing for electron microscopy. As a result, the organization of glandular cells was well-preserved and quite similar to freshly excised rat gastric mucosa for at least 2 h after isolation. Immunohistochemistry of H+/K+-ATPase demonstrated a translocation of the enzyme from the cytoplasm to the apical membrane associated with histamine-stimulation. In cimetidine-treated mucosa, most of the parietal cells were morphologically in the resting state, showing numerous tubulovesicles in their cytoplasm. In contrast, histamine-stimulated parietal cells exhibited well-developed intracellular canaliculi lined with long microvilli.Further study should be required to clarify the functional morphological dynamics of gastric gland.

冷冻固定目前被认为是保存生物标本优良结构和抗原性的最佳初始固定步骤。为了阐明胃壁细胞的功能转化，我们建立了离体胃粘膜的体外实验模型。本研究采用组胺刺激或西咪替丁抑制顶叶细胞分泌酸性物质，并在光镜下采用骤降冷冻或电镜下采用高压冷冻对样品进行冷冻。结果表明，在分离后至少2小时内，腺细胞的组织结构保存完好，与新鲜切除的大鼠胃粘膜非常相似。H+/K+- atp酶的免疫组化显示，该酶从细胞质向顶膜的易位与组胺刺激有关。在西咪替丁处理的粘膜中，大多数壁细胞形态学上处于静息状态，细胞质中有大量的小管泡。相比之下，组胺刺激的壁细胞显示出发育良好的细胞内小管，内衬长微绒毛。胃腺的功能形态动力学有待进一步研究。

项目成果

Aggregation and loss of cytokeratin filament networks inhibit golgi organization in liver-derived epithelial cell lines

• DOI: 10.1002/cm.10152
• 发表时间: 2004-01-01
• 期刊: CELL MOTILITY AND THE CYTOSKELETON
• 作者: Kumemura, H;Harada, M;Sata, M
• 通讯作者: Sata, M

The application of 10 μm-thin stainless foil to a new assembly of the specimen carrier of high pressure freezing.

10μm薄不锈钢箔在高压冷冻标本载体新型组装中的应用

• 发表时间: 2005
• 期刊: J Electron Microsc (in press)
• 作者: Sawaguchi A;Ide S;Suganuma T
• 通讯作者: Suganuma T

Externalization and recognition by macrophages of large subunit of eukaryotic translation initiation factor 3 in apoptotic cells.

• DOI: 10.1016/j.yexcr.2005.05.006
• 发表时间: 2005-09
• 期刊: Experimental cell research
• 影响因子: 3.7
• 作者: Y. Nakai;A. Shiratsuchi;J. Manaka;H. Nakayama;K. Takio;Jian-Ting Zhang;T. Suganuma;Y. Nakanishi

The cryofixation of isolated rat gastric mucosa provides new insights into the functional transformation of gastric parietal cells:: an in vitro experimental model study

• DOI: 10.1679/aohc.68.151
• 发表时间: 2005-09-01
• 期刊: ARCHIVES OF HISTOLOGY AND CYTOLOGY
• 作者: Sawaguchi, A;Aoyama, F;Suganuma, T
• 通讯作者: Suganuma, T

Regeneration of gastric mucosa during ulcer healing follows ontogenetic course of rat fundic glands.

溃疡愈合过程中胃粘膜的再生遵循大鼠胃底腺的个体发育过程。

• 发表时间: 2004
• 期刊: Virchows Arch 445
• 作者: Nagaike R;Sawaguchi A;Kawan J;Aoyama F;Oinuma T;Suganuma
• 通讯作者: Suganuma