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ERM proteins and Odf2 as organizers for apical membranes

ERM 蛋白和 Odf2 作为顶膜的组织者

基本信息

批准号：
17370070
负责人：
TSUKITA Sachiko
金额：
$ 8.9万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17370070/
关键词：
cytoskeleton cell polarity cell membrane actin filament cell adhesion ERM proteins primary cilia Odf2 細胞接着中心体エズリン胃酸プライマリーシリア HK-ATPase

项目摘要

1.Preparation and analysis of ERM protein knockout mice : Analysis of knockout mice clarified that radixin maintains the function of the bilirubin transporter, MRP2, by anchoring it to the luminal surface of capillary bile ducts in the liver. In knockout mice, MRP2 cannot function, and induces Dubin-Johnson syndrome-type hyperbilirubinemia. In this study, we showed the possibility of individual histological and hematological differences in this syndrome reflecting genetic background-associated differences in the basolateral surface MRP3 expression level.2.Analysis of the composition of cell adhesion apparatus : We established a preparation method of a concentrated cell adhesion apparatus fraction from a capillary bile duct fraction prepared from the liver in 1989, and identified major proteins composing the apparatus. In this study, we identified many novel components of the cell adhesion apparatus, which was not possible using previous methods, by taking advantage of the marked progre … More ss in mass spectrometry. Many of these tagged proteins were localized in the cell adhesion apparatus, as expected. We prepared antibodies against these proteins, and RNAi experiments in cultured cells are now underway. We are planning to investigate how these proteins coordinate in their roles in intercellular adhesion.3.Analysis of centrosome component protein, Odf2, knockout mice : In Odf2 conditional knockout mice, cell division may not be affected, and only the primary cilium may organ-specifically disappear. Using these mice, the role of primary cilia may be analyzed in vivo, which has remained unclear for a long time. Odf2 is a component of outer dense fibers (ODF) of sperms, and its disappearance causes male infertility. Since only a slight decrease in the expression caused infertility, preparation of conditional knockout mice was difficult, but we finally established a heterozygote generation. Since an unexpected function of the primary cilium was suggested at the cell culture level, analysis of individual animals may provide significant information. Less

1. ERM蛋白敲除小鼠的制备和分析：敲除小鼠的分析阐明，根毒素通过将胆红素转运蛋白MRP 2锚定到肝脏中毛细胆管的管腔表面来维持其功能。在敲除小鼠中，MRP 2不能发挥功能，并诱导Dubin-Johnson综合征型高胆红素血症。在这项研究中，我们显示了这种综合征个体组织学和血液学差异的可能性，反映了基底外侧表面MRP 3表达水平的遗传背景相关差异。2.细胞粘附装置组成分析：我们在1989年建立了从肝脏制备的毛细胆管部分制备浓缩细胞粘附器部分的方法，并确定了组成该装置的主要蛋白质。在这项研究中，我们利用细胞粘附装置的显着进展，鉴定了许多新的细胞粘附装置组件，这是使用以前的方法不可能的。 ...更多信息质谱中的ss。正如预期的那样，这些标记的蛋白质中的许多定位在细胞粘附装置中。我们制备了针对这些蛋白质的抗体，目前正在培养细胞中进行RNAi实验。3.中心体组分蛋白Odf 2基因敲除小鼠的分析：在Odf 2基因敲除小鼠中，细胞分裂可能不受影响，只有初级纤毛可能器官特异性消失。使用这些小鼠，初级纤毛的作用可以在体内进行分析，这在很长一段时间内仍然不清楚。ODF 2是精子外致密纤维（ODF）的一个组成部分，它的消失导致男性不育。由于只有轻微的表达下降导致不育，条件性基因敲除小鼠的制备是困难的，但我们最终建立了杂合子一代。由于在细胞培养水平上提示初级纤毛具有意想不到的功能，因此对个体动物的分析可提供重要信息。少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Tricellulin constitutes a novel barrier at tricellular contacts of epithelial cells.

DOI：
10.1083/jcb.200510043
发表时间：
2005-12-19
期刊：
The Journal of cell biology
影响因子：
0
作者：
Ikenouchi J;Furuse M;Furuse K;Sasaki H;Tsukita S;Tsukita S
通讯作者：
Tsukita S

Achlorhydria by ezrin knockdown: defects in the formation/expansion of apical canaliculi in gastric parietal cells.

DOI：
10.1083/jcb.200410083
发表时间：
2005-04-11
期刊：
The Journal of cell biology
影响因子：
0
作者：
Tamura A;Kikuchi S;Hata M;Katsuno T;Matsui T;Hayashi H;Suzuki Y;Noda T;Tsukita S;Tsukita S
通讯作者：
Tsukita S

Odf2-deficient mother centrioles lack distal/subdistal appendages and the ability to generate primary cilia