Identification of differentially expressed genes of primary spermatocyte against round spermatid isolated from human testis using the laser capture microdissection technique

作     者：S.S.KOIDE 

作者机构：1230 York Avenue Center for Biomedical Research Population Council NY 10021 New York USA 

出 版 物：《Cell Research》 (细胞研究(英文版))

年 卷 期：2004年第14卷第6期

页      面：507-512页

核心收录：

学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 1001[医学-基础医学(可授医学、理学学位)] 10[医学] 

基　　金：国家自然科学基金(30070173and30240019) 国家高技术研究发展计划(863计划)(2001AA221131) 国家重点基础研究发展计划(973计划)(2002BA711A01) 

主　　题：laser capture microdissection suppressive subtractive hybridization spermatogenesis cytochrome c oxidase Ⅱ humanin. 

摘      要：The method of laser capture microdissection (LCM) combined with suppressive subtractive hybridization (SSH) was developed to isolate specific germ cells from human testis sections and to identify the genes expressed during differentiation and development. In the present study, over 10,000 primary spermatocytes and round spermatid cells were successfully isolated by LCM. Using the cDNAs from primary spermatocytes and round spermatids, SSH cDNAs library of primary spermatocyte-specific was constructed. The average insert size of the cDNA isolated from 75 randomly picked white clones was 500 bp, ranging from 250 bp to 1.7 kb. Using the dot-blot method, a total of 421 clones were examined, resulting in the identification of 390 positive clones emitting strong signals. Partial sequence of cDNAs prepared from each clone was determined with an overall success rate of 84.4%. Genes encoding cytochrome c oxidase II and the rescue factor-humanin were most frequently expressed in primary spermatocytes, suggesting th