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Role of cell adhesion molecules during net formation in Hydorodictyon reticulatum

细胞粘附分子在网状水藻网形成过程中的作用

基本信息

批准号：
13640663
负责人：
HATANO Kyoko
金额：
$ 1.98万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13640663/
关键词：
Hydrodictyon reticulatum net formation cell adhesion microtubules zoospores

项目摘要

A monoclonal antibody that was raised against a homogenate of zoospores recognized cell adhesion molecules localized to the contact sites between zoospores. The molecule was constantly expressed in zoospores during net formation and decreased after the completion of the adhesion between zoospores. The subcellular localization of the molecule during net formation was analyzed by indirect immunofluorescence microscopy. The peripheral regions of the spherical zoospores and the contact regions of the polyhedral zoospores were stained brightly. Immediately after cessation of the movement of polyhedral zoospores, the molecule was located on the closely apposed surfaces of two cells. In order to examine the precise spatial distribution of the molecule, zoospores and net cells were double-stained with this monoclonal antibody and a monoclonal antibody against chicken β-tubulin. The molecule closely colocalized with the microtubule bundles at the contact regions of the polyhedral zoospores and the net cells. Colchicine-treated zoospores could not form hexagonal nets, then the peripheral regions of zoospores stained uniformly. Immunoelectron microscopy revealed the presence of gold particles on a Golgi stack and the plasma membrane in polyhedral zoospores and the contact regions of net cells. I treated isolated zoospores with this monoclonal antibody. Treatment with the antibody inhibited formation of hexagonal nets. These results suggest that localization of the molecule participate in the determination of the contact sites of zoospores and that the microtubule bundles are involved in the localization of the molecule during hexagonal net formation.

一种针对游动孢子匀浆的单抗可识别定位于游动孢子之间接触点的细胞黏附分子。该分子在游动孢子成网过程中持续表达，并在游动孢子之间完成黏附后下降。用间接免疫荧光显微镜分析该分子在网络形成过程中的亚细胞定位。球形游动孢子的周边区和多面体游动孢子的接触区染色较深。在多面体游动孢子停止运动后，分子立即位于两个细胞紧密相对的表面上。用该单抗和抗鸡β-微管蛋白的单抗对游动孢子和网状细胞进行双重染色，以确定分子的精确空间分布。在多面体游动孢子和网状细胞的接触区，分子与微管束紧密共存。秋水仙素处理的游动孢子不能形成六角形的网状，游动孢子的周边区域染色均匀。免疫电子显微镜显示，在高尔基体堆积、多角体游动孢子的质膜和网状细胞的接触区均有金颗粒存在。我用这种单抗处理分离的游动孢子。用该抗体处理可抑制六角网的形成。这些结果表明，分子的定位参与了游动孢子接触位置的确定，而微管束参与了六角形网状形成过程中分子的定位。