摘要
在Balb/c小鼠精子发生过程中,许多基因都具有严格的时空表达特性。实验利用半定量RT-PCR验证了12个小鼠精子发生相关基因的组织学分布,采用SYBR GreenⅠ荧光定量PCR分析了它们在不同发育阶段生精细胞中的差异表达。结果显示,所有基因仅在睾丸组织中高表达;Prm1、Prm2、Tnp1、Tnp2在长形精子细胞中的表达水平最高,分别是粗线期精母细胞阶段的1.9、2.8、3.2和2倍;Dnajb3呈上调表达,在长形精子细胞中的含量是粗线期精母细胞阶段的2.5倍;Akap4在长形精子细胞阶段的表达水平尤为突出,是粗线期精母细胞的5.5倍;Spata3和Spata4在圆形及长形精子细胞中的表达量相近,分别是粗线期精母细胞阶段的3倍和1.5倍;hils1和Tex24在圆形精子细胞阶段的表达水平最高,分别是粗线期精母细胞阶段的1.9和1.4倍;Spag4l和Papolb从粗线期精母细胞到长形精子细胞阶段呈明显的下调表达,分别下降了45%和34%。结果提示,被检测的基因具有明显的阶段特异性表达特征,为深入研究这些基因在小鼠精子发生过程中的作用提供了新资料,同时也为荧光定量PCR技术在精子发生相关基因定量表达研究中的可行性提供了充分例证。
Many testis-specific and stage-specific genes are required for mouse spermatogencsis. In this study, semi- quantitative RT-PCR was used to confirm the tissue distribution of 12 mouse testicular genes, and fluorescence quantitative PCR labeled with SYBR Green I was applied to demonstrate the stage-specific expression of candidate genes at different spermatogenic stages. The results showed that all the genes examined were highly expressed in testis. The expression of Prm1, Prm2, Tnp1, and Trip2 was predominant in elongating spermatids, 1.9-, 2.8-, 3.2-, and 2-fold increase as compared with that in pachytene spermatocytes, respectively. Dnajb3 expression was upregulated 2.5-fold from pachytene spermatocytes to elongating spermatids. Akap4 was strongly expressed at elongating spermatid stage, with 5.5-fold increase than that in pachytene spermatocytes. Both Spata3 and Spata4 were equally expressed in round and elongating spermatids, 3- and 1.5-fold increase as compared with pachytene spermatocytes, respectively. The expression levels of hilsl and Tex24 were the highest in round spermatids, with 1.9- and 1.4-fold increase from pachytene spermatocytes, respectively. Spag41 and Papolb were down-regulated from pachytene spermatocytes to elongating spermatids, decreasing 45 % and 34 %, respectively. The results not only demonstrate the stage-specific expression of these genes, but also provide new data for further investigations into the functions of these specific genes during spermatogenesis.
出处
《动物学杂志》
CAS
CSCD
北大核心
2009年第1期39-46,共8页
Chinese Journal of Zoology
基金
国家自然科学基金青年项目(No.30700911)
关键词
精子发生
荧光定量PCR
小鼠
生精细胞
Spermatogenesis
Fluorescence quantitative PCR
Mouse
Spermatogenic cell
