权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Molecular Cell Physiological Study on Paneth Cell Function

潘氏细胞功能的分子细胞生理学研究

基本信息

批准号：
10470012
负责人：
OKADA Yasunobu
金额：
$ 8.58万
依托单位：
National Institute for Physiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470012/
关键词：
Paneth cell ClィイD1-ィエD1 CFTR cyclic AMP patch-clamp

项目摘要

It is widely accepted that the small internal crypt is a ClィイD1-ィエD1 secreting epithelium where CFTR CIィイD1-ィエD1 channels are thought to mediate cyclic AMP-dependent CIィイD1-ィエD1 exit at the luminal membrane. Actually, abundant expression of CFTR mRNA in the small intestine has been shown by in situ hybridization. Paneth cells, which locate at the crypt base, have been implicated as components of the mucosal immunity and host defense by secreting anti-microbial substances, such as crypidins and lysozymes. Recently, the channel-forming crypidins have been reported to serve an apically localized C1ィイD1-ィエD1 conductance, thereby contributing to the intestinal CIィイD1-ィエD1 secretion. However, there has been no investigation as to whether Paneth cells express CFTR C1ィイD1-ィエD1 channels. In the present study, whole-cell patch clamp was first applied to Paneth cells in crypts isolated from guinea pig small intestine. Prominent activation of ClィイD1-ィエD1 currents could be observed only after stimulation with VIP or dibutyryl cyclic AMP plus forskolin. The current characters, including the current-voltage relationship, anion selectivity and DIDS-insensitivity, were phenotypically similar to those of CFTR CIィイD1-ィエD1 channel. However, both single-cell RT-PCR analysis and immunocytochemical staining failed to provide relevant evidence for molecular expression of the CFTR mRNA and protein. Thus, it is concluded that Paneth cells express cyclic AMP-activated CI' channels, but CFTR would not be the molecular identity of the ClィイD1-ィエD1 channel in Paneth cells of the guinea pig small intestine.

人们普遍认为，小内隐窝是一种CIィイD1-ィエD1分泌上皮，其中CFTRCIィイD1-ィエD1通道被认为介导依赖于cAMP的CIィイD1-ィエD1在管腔膜上的存在。事实上，原位杂交显示CFTRmRNA在小肠中有丰富的表达。位于隐窝底部的Paneth细胞通过分泌抗微生物物质，如隐孢子素和溶菌酶，被认为是粘膜免疫和宿主防御的组成部分。最近，通道形成的隐素被报道起到顶端定位的C_1ィイD_1-ィエD_1电导作用，从而促进肠道CIィイD_1-ィエD_1的分泌。然而，目前还没有关于Paneth细胞是否表达CFTRC1ィイD1-ィエD1通道的研究。本研究首次将全细胞膜片钳技术应用于豚鼠小肠隐窝的Paneth细胞。只有在血管活性肠肽或二丁酰环磷酸腺苷加福司可林刺激后，才能观察到氯ィイD1-ィエD1电流的显著激活。电流特性，包括电流-电压关系、阴离子选择性和DDS不敏感度，与CFTRCIィイd1-ィエd1通道相似。然而，单细胞RT-PCR分析和免疫细胞化学染色均未能为CFTRmRNA和蛋白的分子表达提供相关证据。因此，我们认为豚鼠小肠潘氏细胞表达cAMP激活的CI‘通道，但cftr不是豚鼠小肠潘氏细胞中CL-ィイD1-ィエD1通道的分子特征。