背景:精子发生过程受许多特异分子及细胞间作用的严格调节,包括染色体结构变化及一系列特定基因程序性表达调控。长期以来由于缺乏合适的体内和体外研究模型,精子发生分子机制的研究进展较慢,尤其缺乏对关键调节基因的认识。目的:筛选与精子发生相关的基因,并分析其表达特点。方法:将4,9,18,35,54d和6月龄小鼠睾丸组织cDNA探针与Affymetrix全基因组芯片进行杂交,筛选出差异表达的基因。然后通过反转录-聚合酶链反应分析差异表达基因在小鼠睾丸不同发育阶段中的表达。结果与结论:对Affymetrix全基因组芯片杂交结果分析后,筛选得到1个差异表达杂交点,通过在NCBI与小鼠全基因组序列Blast分析可知该差异表达基因是Tpap基因。该基因在4,9,18,35,54d和6月龄小鼠睾丸中杂交信号校正值分别是4.4(A)、12.9(A),262.4(P)、1136.7(P)、1617.5(P)和1128(P),表明4,9d小鼠睾丸中该基因无表达,18d小鼠开始表达,RT-PCR结果表明小鼠Tpap基因在小鼠9d及之前的睾丸中没有表达,在18d睾丸后开始表达,与基因芯片分析相一致。结果表明,Tpap基因为小鼠年龄依赖性表达基因,小鼠Tpap的表达与小鼠精子发生的过程有很强的一致性,推测该基因在哺乳动物精子发生中起重要作用。
BACKGROUND:These serial processes for forming male gametes are basically controlled by the programmed expression of a number of stage-specific genes.However,many aspects of the mechanisms of spermatogenesis have remained elusive because of a lack of suitable in vitro or in vivo models.OBJECTIVE:To screen genes involved in spermatogenesis,and to analyze its expression characteristics.METHODS:Testes cDNA samples from Balb/C mice of different postnatal days(4,9,18,35,54 days and 6 months,respectively)were hybridized with mouse whole genome Affymetrix chip to screen the testis-ralated genes.The characteristics of the selected genes were analyzed by various bioinformatics tools.RT-PCR was used here to identify the expression of the selected genes in mice testis.RESULTS AND CONCLUSION:The Affymetrix chip probe of mouse Tpap was graduated higher expression with developmental stages of mouse testis.The scaling hybridization signal intensities of the tested testis on days 4,9,18,35,54,and 6 months of postnatal were 4.4(Absent expression,A),12.9(A),262.4(Present expression,P),1136.7(P),1617.5(P)and 1128(P),respectively.These results indicated that the expression of mouse Tpap wasn't detected on days 4 and 9,but was detected on days 18,35,54,and 6 months of mouse testis in our Affymetrix chip analysis.By combination with the RT-PCR analysis of mouse Tpap,we observed mouse Tpap began to express at the age of day 18 in mouse.Tpap is an age-dependent gene in mouse testis.The expression of Tpap corresponds to the appearance of spermatids of mice and indicates that Tpap may have an important role in male mammalian spermatogenesis.