生殖细胞及体细胞相关标记蛋白在小鼠胚胎雌雄生殖嵴的表达差异研究

Comparative study of gonadogenesis in fetal stage between male and female mice

目的比较小鼠胚胎雌雄生殖嵴发育过程中的生殖细胞及体细胞相关标记蛋白的表达差异。方法运用免疫荧光技术分别在雌、雄小鼠12.5、13.5、14.5、18.5dpc(days postcoitum)及1dpp(days postpartum)的生殖嵴中标记DDX4(DEAD Asp-Glu-Ala-Asp box polypeptide 4)、OCT4(POU domain,class 5,transcription factor 1)、GATA4(Transcription factor GATA-4)、FOXL2(Forkhead box protein L2)、SOX9(Transcription factor SOX-9)、γ-H2AX(The phosphorylated form ofHistone H2AX)及磷酸化RB1(Retinoblastoma-associated protein)(P-RB1)。观察这些蛋白质在小鼠胚胎的不同时间点雌雄生殖嵴中的表达变化情况。结果 DDX4在早期胚胎各阶段雌、雄生殖嵴的生殖细胞呈现持续的表达;FOXL2和SOX9分别在早期胚胎雌、雄生殖嵴的体细胞持续表达;而其它标记蛋白(OCT4、GATA4、γ-H2AX和P-RB1)在早期胚胎不同阶段却呈现差异表达模式,OCT4在雄性生殖嵴的生殖细胞各个阶段均有表达,而在雌性生殖嵴14.5dpc已不表达;GATA4在雄性生殖嵴的体细胞各个阶段均有表达,但在雌性生殖嵴18.5dpc已不表达;γ-H2AX在雌性生殖嵴18.5dpc已不表达;P-RB1在雄性生殖嵴13.5dpc的生殖细胞开始不表达,而在雌性生殖嵴18.5dpc开始不表达。结论不同细胞组分功能蛋白的特异性表达模式对于早期胚胎生殖嵴的发育和分化起到重要作用。

Objective To compare the main biological processes in the development of gonads between male and female mice.Methods Female and male gonads collected from 12.5,13.5,14.5,18.5dpc and 1dpp were labeled by markers DDX4,OCT4,GATA4,FOXL2,SOX9,γ-H2 AX and phospho-RB1(PRB1)to observe the expression patterns of these markers at different time points between male and female mouse embryos.Results DDX4 was found to be expressed consistently at germ cells in every time point of female and male gonads,FOXL2 and SOX9were distributed persistently in somatic cellsof female and male gonads,respectively,while the other markers(OCT4,GATA4,γ-H2 AX and P-RB1)showed differential expression among these stages between male and female embryos.OCT4 was expressed consistently at germ cells in every time point of male gonads,while no signal was detected from 14.5dpc in females;GATA4was expressed consistently at somatic cells in every time point of male gonads,while no signal was detected from 18.5dpc in females;γ-H2 AX showed no expression from 18.5dpc in female;P-RB1 ceased to express in germ cells from 13.5dpc in male and 18.5dpc in female gonads.Conclusion The specific expression of functional proteins from different cell lineages may play crucial roles in the gonadal development and differentiation between male and female mouse embryos.