| 摘要: | 組織蛋白是染色質中的基本組成蛋白質，組織蛋白與染色質規則纏繞成nucleosome，成為染色質的最基本單位，包括H2A、H2B、H3、H4及當作為二單位nucleosomes間的linker蛋白H1組織蛋白。組織蛋白與染色質形成類似線上的珠串（beads-on-a-string）構造，可使染色質規律排列成纏繞緊密的染色體結構。本研究是以過去我們在微陣分析（microarray）無精蟲症患者及正常生育者的睪丸組織表現的mRNA表現量分析，發現部分組織蛋白H3/B基因在睪丸組織表現的mRNA表現量有統計上顯著之差異，因此本研究希望針對此H3B基因參與造精過程做進一步深入研究。本研究以bioinformatics search方法進行染色體上序列的比對，檢索正確的基因所在位置及基因序列，並將組織蛋白進行分群結果，選定H3d、H3h2ba、H3fa及H3fb等為研究對象，其中H3fa及H3fb為正控制組的基因，進行此些H3B基因mRNA在老鼠睪丸發育不同階段，H3B基因在睪丸表現情形。研究結果顯示，組織蛋白H3d是屬於造精過程中早期表現的基因；組織蛋白H3h2ba是屬於造精過程中每一階段均會表現的基因；組織蛋白H3fa及H3fb是屬於造精過程晚期表現的基因，本研究且同時分析H1、H2及H4等其他組織蛋白在造精過程中表現情形，以期對於組織蛋白的表現及其參與造精過程的功能與角色有更深入的瞭解。
Mammalian spermatogenesis is a highly ordered process that occurs in mitotic, meiotic and post-meiotic phases. During spermatogenesis, chromatin is restructured, and dramatic changes in histone gene expression are observed. Histones are a major component of chromatin, the protein-DNA complex fundamental to genome packaging, function, and regulation in spermatogenesis . Indeed, during the post-meiotic maturation of male haploid germ cells, or spermiogenesis, histone are replaced by small basic proteins, which in mammals are transition proteins and protamines.The genes of this study included by the following databank: 1.The meiotic or postmeiotic spermatogenic cells by Nikolaus S. et al.2.Mouse spermatogenesis genes in SAGE 3.Genes of testis bank4. the mouse UniGene libraryCombining of the above mouse spermatogenesis genes banks by raw data, the 20 histone candidate genes were selected. Specific primers of 20 histone genes were designed by roche software. Total RNA from 5-d-old (5d) , 10-d-old (10d) , 15-d-old (15d) , 20-d-old (20d) ,35-d-old (35d) , and 47-d-old (47d) testes were isolated and determined by RT-PCR. The expression of these 20 histone genes were determined by semi-quantitative RT-PCR among normal, MA, and SCOS patients. The 20 andidate genes involved in spermatogenesis will determined by RNA immunoprecipitatin and EMSA. We hope to identify and characterize the histone genes expression patterns in spermatogenesis. The study may help to clarify the role of histone gene in human spermatogenesis. |
