Guo Rui, Yu Zuoren, Guan Jikui, Ge Yehua, Ma Jing, Li Sai, Wang Shali, Xue Shepu, Han Daishu
Department of Cell Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Mol Reprod Dev. 2004 Mar;67(3):264-72. doi: 10.1002/mrd.20026.
Spermatogenesis occurs in successive mitotic, meiotic, and post-meiotic phase, and involves a number of unique processes including meiosis and dramatic morphological changes. The unique differentiation mechanisms of spermatogenesis suggest the existence of germ-cell-specific molecules. The most straight forward strategy to elucidate differentiation mechanisms is to identify and characterize differentiation-specific molecules and their associated genes in germ cells. However, only a few genes specifically involved in spermatogenesis have been studied. In the present study, six different types of spermatogenic cells (primitive type A spermatogonia, type B spermatogonia, preleptotene spermatocytes, pachytene spermatocytes, round spermatids, and elongating spermatids) were isolated from Balb/c mice testes using velocity sedimentation and Atlas cDNA arrays containing 1,176 known mouse genes were used to determine the gene expression profiles of the spermatogenic cells. The expression of 260 genes were detected in six different stages of spermatogenic cells and a number of genes showed differential expression. The 23 differentially expressed genes were further analysed by reverse transcription polymerase chain reaction (RT-PCR) for their stage-specific and tissue-specific expression characteristics. Based on the results of RT-PCR, six genes highly express in both primitive type A and type B spermatogonia, four genes up-regulate in type B spermatogonia, two genes up-regulate in spermatocytes, two genes up-regulate in spermatids, three genes express constantly from primitive A spermatogonia to elongating spermatids, two genes express constantly from primitive A spermatogonia to round spermatids, two genes do not change in their expression during spermatogenesis, two genes can be detected highly in adult testis, but are undetectable in spermatogenic cells. The tissue-specific expression characteristics of the 23 genes showed that some of them specifically expressed in testes or other tissues. These data provide new information for further studies into spermatogenesis-related genes and may lead to the identification of genes with potential relevance to the differentiation of spermatogenic cells. In addition, some of these genes could be considered to be used as the molecular markers for different stages of spermatogenic cells.
精子发生过程包括连续的有丝分裂、减数分裂和减数分裂后阶段,涉及许多独特的过程,包括减数分裂和显著的形态变化。精子发生独特的分化机制表明存在生殖细胞特异性分子。阐明分化机制最直接的策略是鉴定和表征生殖细胞中分化特异性分子及其相关基因。然而,仅有少数几个特异性参与精子发生的基因得到了研究。在本研究中,利用速度沉降法从Balb/c小鼠睾丸中分离出六种不同类型的生精细胞(原始A型精原细胞、B型精原细胞、前细线期精母细胞、粗线期精母细胞、圆形精子细胞和伸长精子细胞),并使用包含1176个已知小鼠基因的Atlas cDNA阵列来确定生精细胞的基因表达谱。在生精细胞的六个不同阶段检测到了260个基因的表达,并且许多基因表现出差异表达。通过逆转录聚合酶链反应(RT-PCR)对23个差异表达基因的阶段特异性和组织特异性表达特征进行了进一步分析。基于RT-PCR的结果,六个基因在原始A型和B型精原细胞中均高表达,四个基因在B型精原细胞中上调,两个基因在精母细胞中上调,两个基因在精子细胞中上调,三个基因从原始A型精原细胞到伸长精子细胞持续表达,两个基因从原始A型精原细胞到圆形精子细胞持续表达。两个基因在精子发生过程中表达无变化,两个基因在成年睾丸中可高表达,但在生精细胞中未检测到。这23个基因的组织特异性表达特征表明,其中一些基因在睾丸或其他组织中特异性表达。这些数据为进一步研究精子发生相关基因提供了新信息,并可能有助于鉴定与生精细胞分化潜在相关的基因。此外,这些基因中的一些可被视为不同阶段生精细胞的分子标记。