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目的:研究精子顶体小泡蛋白-1(ACRV1)在小鼠睾丸组织发育过程中的表达特征。方法:将4 d、9 d、18 d、35 d、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ACRV1基因。采用RT-PCR方法检测ACRV1基因在小鼠不同日龄和不同组织中的表达情况,采用免疫组织化学方法检测ACRV1蛋白在小鼠睾丸组织中的定位。结果: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网站与小鼠全基因组序列Blast分析可知该差异表达基因是ACRV1基因。RT-PCR结果表明ACRV1 mRNA呈小鼠睾丸特异性表达,在出生31 d小鼠睾丸开始高表达,在成年前达到高峰。ACRV1蛋白主要定位在睾丸圆形精子和长形精子细胞。结论:ACRV1基因存在发育的表达调控,为小鼠年龄依赖性表达基因,其表达与小鼠精子发生的过程有很强的一致性,且具有睾丸特异性表达的特征,因此推测该基因可能在精子发生中具有关键作用。
Objective: To study the expression of sperm acrosome protein 1 (ACRV1) during the development of mouse testis. METHODS: The cDNA probes of testis tissue from 4 days, 9 days, 18 days, 35 days, 54 days and 6 months old mice were hybridized with Affymetrix genome-wide cDNA microarray. The differentially expressed ACRV1 gene was screened out. The expression of ACRV1 gene in different age and different tissues of mice was detected by RT-PCR, and the localization of ACRV1 protein in mouse testes was detected by immunohistochemistry. RESULTS: After analysis of Affymetrix whole-genome chip hybridization results, one differential expression hybridization point was screened. According to Blast analysis on the NCBI website and mouse whole genome sequence, the differentially expressed gene was ACRV1 gene. RT-PCR results showed that ACRV1 mRNA was specifically expressed in the testes of mice, and began to be highly expressed in testis of mice on day 31 and reached a peak before adulthood. ACRV1 protein is mainly located in testicular round sperm and sperm cells. CONCLUSION: ACRV1 gene is regulated in development and age-dependent. The expression of ACRV1 gene is highly consistent with the process of mouse spermatogenesis and characterized by testicular-specific expression. Therefore, Spermatogenesis plays a key role.