小鼠睾丸支持细胞标志蛋白表达的研究

发布时间：2018-07-20 13:53

【摘要】：目的:探寻小鼠睾丸支持细胞不同发育阶段和功能状态下的特异性标志蛋白。方法:通过对小鼠睾丸组织进行免疫组织化学染色和实时定量(real-time)RT-PCR,观察受精后16 d胚胎(E16)到出生后6周的雄性小鼠睾丸组织蛋白和m RNA表达。结果:雄激素受体(AR)在出生后1周、2周、6周小鼠的睾丸支持细胞、外周小管肌样细胞、间质细胞细胞核内都有表达,且表达量逐渐增强,但在E16不表达;Wilms肿瘤基因(WT1)在E16、出生后1周、2周、6周的睾丸支持细胞细胞核内表达,但不表达于其它细胞;苗勒氏管抑制素(AMH)在E16、出生后2周的睾丸支持细胞胞质中表达,而在出生后6周不表达;β-链蛋白(β-catenin)和胞质紧密粘连蛋白-1(Zo-1)是血睾屏障特异性蛋白,出生后2周时它们分布于整个曲细精管,而出生后4周时规则地分布于靠近外壁的一圈,此时它们的血睾屏障已经完全建立。Real-time PCR分析睾丸组织m RNA表达,佐证了这些结果。结论:WT1可以作为支持细胞的特异性标志,而AMH则标记不成熟的睾丸支持细胞;AR单独不适合作为睾丸支持细胞的标志物,但与其他标志物结合,可能可区分睾丸支持细胞的成熟与否。而通过β-catenin和Zo-1染色,可以判断血睾屏障是否完全形成,为支持细胞是否成熟提供判断标记。
[Abstract]:Objective: to explore the specific marker proteins of mouse testicular Sertoli cells in different developmental stages and functional states. Methods: immunohistochemical staining and real-time quantitative (real-time) RT-PCR were used to detect the expression of protein and mRNA in testis of male mice from 16 days after fertilization (E16) to 6 weeks after birth. Results: the expression of androgen receptor (AR) in testicular Sertoli cells, peripheral tubuloid cells and interstitial nuclei of mice at 1 week, 2 weeks and 6 weeks after birth was gradually increased. However, the Wilms tumor gene (WT1) was not expressed in the Sertoli nuclei of testis in E16, but not in other cells, and AMH was expressed in the cytoplasm of Sertoli cells 2 weeks after birth. 尾 -catenin and cytoplasmic compact adhesion protein 1 (Zo-1) are blood-testis barrier specific proteins, which are distributed throughout the seminiferous tubules at 2 weeks after birth, and regularly in a circle near the outer wall at 4 weeks after birth. By this time, their blood-testis barrier had been completely established. Real-time PCR was used to analyze the expression of mRNA in testis, which confirmed these results. Conclusion: WT1 can be used as a specific marker for Sertoli cells, and AMH for immature Sertoli cells alone is not suitable as a marker of testicular Sertoli cells, but combined with other markers. It may be possible to distinguish the maturation of testicular Sertoli cells. 尾 -catenin and Zo-1 staining can be used to determine whether the blood-testis barrier is completely formed and provide a marker for the maturation of Sertoli cells.
【作者单位】：厦门大学物理与机电工程学院;厦门大学药学院;
【基金】：国家自然科学基金资助(基金编号31171375)
【分类号】：R339.21

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