Tpap基因在小鼠睾丸中的表达

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4,9,18,35,54 d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全基冈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)、1 136.7(P)、1 617.5(P)和1 128(P),表明4,9 d小鼠睾丸中该基因无表达,18 d小鼠开始表达,RT-PCR结果表明小鼠Tpap基因在小鼠9 d及之前的睾丸中没有表达,在18 d睾丸后开始表达,与基因芯片分析相一致.结果表明,Tp印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.

Expression of Tpap gene in mouse testis

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.