探討新穎蛋白LRWD1 在睪丸與精子上所扮演的角色

CNU IR > Pharmacy and Science > Dept. of Biotechnology (including master's program) > Dissertations and Theses > Item 310902800/9688

Please use this identifier to cite or link to this item: https://ir.cnu.edu.tw/handle/310902800/9688

Title:	探討新穎蛋白LRWD1 在睪丸與精子上所扮演的角色 Investigating the role of the novel LRWD1 protein in testis and spermatozoa
Authors:	林揚斌 Yang-Ben Lin
Contributors:	鄧燕妮嘉南藥理科技大學：生物科技研究所
Keywords:	尾巴中心體造精過程 tail Spermatogenesis Centrosome
Date:	2008
Issue Date:	2008-12-29 15:21:14 (UTC+8)
Abstract:	LRWD1 (Leucine-Rich repeats and WD repeat domain containing 1)是在睪丸組織裏大量表現的基因。人類的LRWD1基因位於染色體7q22.1，整個基因體含有15個表現子（exon），蛋白質分子量約70kDa。利用SMART網站（http://smart.embl-heidelberg.de/）預測LRWD1蛋白整個基因體包含的重要domain，發現LRWD1蛋白有一個白胺酸多重複序列（Leucine-Rich repeats, LRR）與三個色胺酸-天門冬胺酸（trypotophan-aspartic acid, WD40）的motif。經由西方墨點法分析，LRWD1為睪丸組織主要表現的蛋白；利用睪丸組織切片來做免疫組織化學染色，發現LRWD1蛋白表現在減數分裂時期。免疫螢光染色分析人類與老鼠的精子細胞發現LRWD1表現在中段（middle piece）與尾部（tail）上，在中段上的表現位置與中心體的蛋白centrin 和γ-tubulin 的位置相同，故LRWD1 蛋白疑似為中心體蛋白（Centrosomal protein）。從睪丸組織分離的成熟與未成熟的精細胞，進行免疫螢光染色分析，發現LRWD1可能為老鼠精子上中心體未退化的蛋白。在雙尾型尾部異常的精蟲發現2個由LRWD1表現中心，並由此中心向後延伸出精蟲尾部。LRWD1在不同活動力精蟲的表現經統計分析後發現，活動力較高低與LRWD1的表現有正相關性(r=0.4098; P=0.0006)，也就是說，LRWD1蛋白的表現可能會影響精蟲活動力。 LRWD1 (Leucine-Rich repeats and WD repeat domain containing 1) is a testis-enriched gene. The human LRWD1 gene contains 15 exons and is mapped to chromosome 7q22.1. The protein is predicted to contain one leucine-rich repeats (LRR) and three WD-40 domains by SMART protein prediction web site (http://smart.embl-heidelberg.de/). The Western blot analysis showed the LRWD1 organization display mainly in testis. We also found the protein was mainly expressed in postmeiotic germ cells. Immunofluorescent microscopy study of human and mouse spermatozoa showed strong homogeneous staining in the centrosome region by colocalization with centrin and γ-tubulin and less intense signal in the tail region. In the isolated germ cell, LRWD1 was the non-degenerated centrosomal protein on mouse spermatogenesis. Therefore, LRWD1 was suspected in the functional discussion of sperm morphology and motility. There are two LRWD1 centers, extent two tails, in the two tails defect sperm. A significant positive correlation was seen between LRWD1 expression and sperm motility (r=0.4098; P=0.0006). Meanwhile, LRWD1 expression might directly involve in the function of sperm motility.
Relation:	校內一年後公開，校外永不公開
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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