Molecular structure of sperm flagella in mammal

哺乳动物精子鞭毛的分子结构

• 批准号: 18570059
• 负责人: IIDA Hiroshi
• 金额: $ 2.53万
• 依托单位: Kyushu University
• 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-18570059/
• 关键词: spermatozoa; development and differentiation; molecular recognition; genes; 発生・文化; 鞭毛; 軸糸; Tektin; Spetex

This study is aimed to elucidate the molecular architecture of mammalian sperm flagella with special reference to Tektin4 and Spetx-1 that have been cloned by differential display and yeast two-hybrid technique. We have disclosed by immnunoelectron microscopy that Tektin4 is present at the surface of outer dense fiber (ODF) and Spetex-1 is a component of satellite fibrils. In addition, we showed that Tektin2 and Tektin5 were localized around ODF and the inner surface of mitochondrial sheath, respectively. We also found that Gcl-2 is associated with the cortex of ODF, like Tektin4. We biochemically studied the localization of these molecules by immunoblot analyses using fractionated samples prepared from purified rat spermatozoa, i.e., plasma membrane, axoneme, ODF, and fibrous sheath. The outcome revealed that all of these molecules were found to be detected in 6M urea-extracted fraction (ODF and ODF-related fractions). When Tektin4 and Spetex-1 were co-transfected into COS-7 cells, the two molecules were found to be co-localized in patch-like structures around nuclei of the cells, the observation suggesting that the two molecules could interact each other in the cells. Finally, we performed yeast two-hybrid screening to isolate molecules that interact with Tektin2 and Tektin5, which resulted in isolation of several novel molecules. Through study of these molecules, we will make a progress for understanding the molecular architecture of mammalian sperm flagella.

这项研究的目的是阐明哺乳动物精子鞭毛的分子结构，并特别提及Tektin4和Spetx-1，这些tektin4和spetx-1被差分显示和酵母两杂化技术克隆。我们已经通过Immnunoelectron显微镜揭示了Tektin4存在于外部致密纤维（ODF）的表面，而Spetex-1是卫星纤维的组成部分。此外，我们表明Tektin2和Tektin5分别位于ODF和线粒体鞘的内表面。我们还发现GCL-2与ODF的皮质相关，例如Tektin4。我们使用从纯化的大鼠精子（即质膜，轴突，ODF和纤维鞘）制备的分离样品来通过免疫印迹分析来研究这些分子的定位。结果表明，在6M尿素提取的部分（ODF和与ODF相关的分数）中发现了所有这些分子。当将Tektin4和Spetex-1共转染到COS-7细胞中时，发现两个分子在细胞核周围的斑块样结构中共定位，观察结果表明两个分子可以在细胞中相互相互作用。最后，我们进行了与Tektin2和Tektin5相互作用的分离分子的酵母两杂化筛选，从而隔离了几种新型分子。通过研究这些分子，我们将取得进步，以了解哺乳动物精子鞭毛的分子结构。

项目成果

Molecular localization of Spetex-1 in the middle piece of flagellum in rodent spermatozoa

啮齿动物精子鞭毛中段 Spetex-1 的分子定位

• 发表时间: 2006
• 作者: Takane Kaneko;Hiroshi iida
• 通讯作者: Hiroshi iida

Molecular localization fo tektin5 in sperm flagella

精子鞭毛中 tektin5 的分子定位

• 发表时间: 2008
• 作者: Murayama;E.;Yamamoto;E.;Kaneko;T.;Iida;H
• 通讯作者: H

Expression of Mina53, a product of a Myc target gene in mouse testis.

• DOI: 10.1111/j.1365-2605.2005.00572.x
• 发表时间: 2006-04
• 期刊: International journal of andrology
• 作者: M. Tsuneoka;Y. Nishimune;K. Ohta;K. Teye;Hiromitsu Tanaka;M. Soejima;H. Iida;T. Inokuchi;H. Kimu

ラット精子鞭毛の外側粗大線維皮質部に局在するgcl-2タンパク質について

关于定位于大鼠精子鞭毛外粗纤维皮层的gcl-2蛋白

• 发表时间: 2007
• 作者: 飯田 弘、村山 恵美、金子 たかね;瓦田 一彦、栗尾 仁之、佐田 純一、矢野 里枝子
• 通讯作者: 瓦田 一彦、栗尾 仁之、佐田 純一、矢野 里枝子

Strucutural proteins in sperm flagella, Spetex-I and Tektin4

精子鞭毛、Spetex-I 和 Tektin4 中的结构蛋白

• 发表时间: 2008
• 作者: Kaneko;T.;Kawarada;K.;Sata;J.;Iida;H
• 通讯作者: H