Lee Kai-Fai, Yeung William S B, Chow Judy F C, Shum Cathy K, Luk John M
Department of Obstetrics and Gynecology, University of Hong Kong, Pokfulam, China.
Biol Reprod. 2004 Apr;70(4):1010-7. doi: 10.1095/biolreprod.103.022061. Epub 2003 Dec 3.
Spermatogenesis is a complicated process of germ cell differentiation, involving programmatic expression of diverse cell type- and developmental stage-specific genes. To date, the vitamin-A-deficiency (VAD) rats and postnatal rats are two models commonly used to study spermatogenesis. In the present study, we studied the expression of 1185 known genes in the vitamin-A-deficient and retinol-reinitiated spermatogenesis of rat testis using Clontech Atlas rat cDNA expression arrays. The mRNA expression patterns of post-vitamin-A (PVA) testis on Days 15 and 35 were compared with those of the spermatogenic arrested rat testis on Day 0. About 9% (110/1185) of the genes studied were highly expressed. When compared with VAD rat testis on Day 0, 20 and 31 genes were differentially expressed by a factor of twofold or greater on Days 15 and 35, respectively. Four genes (cytochrome P450 17, sulfated glycoprotein 2, protein kinase inhibitor, and cathepsin L) that play important roles in spermatogenesis were selected and their gene expression patterns were confirmed by semiquantitative reverse transcription-polymerase chain reaction. Comparison of the expression patterns of these genes between PVA-VAD and postnatal rat testis in developmentally matched stages revealed substantial differences during the early stages of spermatogenesis. This discrepancy could be caused by either the presence of arrested but mature somatic cells in the PVA-VAD testis that may contribute to a unique gene expression pattern in this model or the direct effect of retinol on spermatogenesis. Therefore, caution is needed in interpreting the gene expression data using the PVA-VAD and postnatal rat models in studying spermatogenesis in rat testes.
精子发生是一个复杂的生殖细胞分化过程,涉及多种细胞类型和发育阶段特异性基因的程序性表达。迄今为止,维生素A缺乏(VAD)大鼠和出生后大鼠是常用于研究精子发生的两种模型。在本研究中,我们使用Clontech Atlas大鼠cDNA表达阵列研究了1185个已知基因在维生素A缺乏和视黄醇重新启动的大鼠睾丸精子发生中的表达。将维生素A缺乏后(PVA)第15天和第35天睾丸的mRNA表达模式与第0天精子发生停滞的大鼠睾丸的表达模式进行比较。所研究的基因中约9%(110/1185)高度表达。与第0天的VAD大鼠睾丸相比,分别在第15天和第35天有20个和31个基因差异表达达两倍或更高倍数。选择了四个在精子发生中起重要作用的基因(细胞色素P450 17、硫酸化糖蛋白2、蛋白激酶抑制剂和组织蛋白酶L),并通过半定量逆转录-聚合酶链反应确认了它们的基因表达模式。在发育匹配阶段比较PVA-VAD和出生后大鼠睾丸中这些基因的表达模式,发现在精子发生早期存在显著差异。这种差异可能是由于PVA-VAD睾丸中存在停滞但成熟的体细胞,可能导致该模型中独特的基因表达模式,或者是视黄醇对精子发生的直接作用。因此,在使用PVA-VAD和出生后大鼠模型解释大鼠睾丸精子发生的基因表达数据时需要谨慎。