精原干细胞增殖和分化阶段小鼠睾丸基因的差异表达

Mouse testicular gene expression pattern differences between spermatogonial stem cell proliferative and differential stages

目的分析精原干细胞增殖和分化阶段小鼠睾丸组织基因表达谱的变化，初步探讨精原干细胞增殖和分化的调控机制。方法48只雄性昆明白小鼠采用间隔24d二次腹腔注射白消安（10mg／kg）制备小鼠精子再生模型，8只作为对照组。根据精子再生过程生精上皮的组织形态学变化以及精原细胞增殖情况选取精原干细胞处于增殖和分化阶段的睾丸组织，运用基因表达谱芯片检测2个阶段的睾丸组织基因表达差异，对差异表达基因进行生物信息学分析。结果检测到睾丸组织差异表达基因911个，上调608个（增殖期／分化期）、下调303个。差异表达基因分别涉及生物学过程、分子功能和分子组成。84个信号通路功能改变差异有统计学意义（P〈0．05），包括Notch和Wnt信号通路。与干细胞相关的差异基因有56个，上调40个、下调16个。部分干细胞的阳性标记物（如Cd9、Stra8、Itgb1、Oct4和Thy1）和部分生长因子（如Fgf2、Csf1和Pdgfa）上调。结论小鼠精原干细胞增殖和分化过程的调控涉及许多基因（分属不同信号通路）的差异表达，这些基因和通路对精原干细胞增殖的作用还有待进一步研究。

Objective To detect the mouse testicular gene expression pattern differences between spermatogonial stem cell （SSC） proliferative and differential stages and study the molecular regulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneaily with two doses of busulfan （10 mg/kg） to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SGC proliferation and differentiation. Bioinformatics analysis was conducted in GO （gene ontology） and KEGG （Kyoto Encyclopedia of Genes and Genones） pathway to describe the potential roles that may play in spermatogonial stem cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical significance （P〈0.05）appeared in 84 KEGG signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among whiela 40 genes were up-regulated, including some stem cell biomarkers（such as Cd9, Stra8, Itgb1, Oct4 and Thy1）and some growth factors（such as Fgf2, Pdgfa and Csf1）. Conclusions The regulation of SSC proliferation and differentiation involves in many differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.