Expression of Crumbs-3(Crb3) in Testis at Different Postnatal Stages of Mouse(Mus musculus) | 小鼠生后不同发育阶段睾丸组织中Crumbs-3(Crb3)的表达

Chinese. 为确定极性调控蛋白Crumbs-3(Crb3)在小鼠(Mus musculus)睾丸组织的定位及其在曲精小管精子发生及间质细胞发育过程中的分布规律，本研究运用常规石蜡切片-HE染色方法和免疫荧光组织化学技术，分别对小鼠生后不同发育阶段睾丸组织结构及Crb3在不同发育时期睾丸组织中的分布进行了研究。结果显示，随着个体发育，曲精小管的直径及生精上皮的厚度不断增大，管腔则由小增大、变小再增大；睾丸曲精小管内生精细胞处于动态的发育变化中，2周龄的生精小管内仅有增殖分裂的数层精原细胞和支持细胞，4周龄时出现各级生精细胞、但无精子形成；之后，继续增殖分裂，至8周管腔内有大量精子形成；并随日龄增加生精细胞密度不断加大；支持细胞的形态随着个体发育逐渐变得清晰而易于辨认；间质细胞则随个体发育体积不断增大、核质比例逐渐下降，而胞质嗜酸性逐渐增强。免疫荧光染色结果显示，Crb3蛋白主要分布在各周龄睾丸曲精小管生精细胞胞膜以及睾丸间质细胞胞膜，且间质细胞上的阳性信号明显强于生精细胞。研究结果提示，Crb3不仅与生精上皮细胞的极性建立和维持有关，而且可能参与睾丸类固醇激素的合成和分泌。

English. The aim of present research is to determine the localization of Crumbs-3(Crb3) in mouse testis and its distribution rules on spermatogenesis in seminiferous tubules and developmental process of Leydig's cells. We studied the histological structure and distribution of Crb3 at different postnatal development stages of mouse(Mus musculus) testis by common paraffin sectioning method stained with Hematoxilin and Eosin method and immunofluorescence histochemistry. The results showed that the diameter and thickness of seminiferous tubules increased，their lumens experienced a process from small to big, then narrowed before widened lastly as individual development. Spermatogenic cells in seminiferous tubules were in dynamic development changes, in which there were only some spermatogonium and Sertoli cells at 2th week of age, then all spermatogenic cells except spermatozoa could be seen at 4th week of age, a great quantity of spermatozoa appeared at 4th week of age after proliferation of spermatogenic cells. Then the density of spermatogenic cells increased as development. The shape of Sertoli cells became clear and easily to be recognized, and the volume of leydig cells and eosinophilic cytoplasm increased gradually, while nuclear-cytoplasmic ratio decreased gradually as development. Crb3 protein were mainly distributed in membrane of both spermatogenic cells and leydig's cells, and the positive reaction in leydig's cells were obviously stronger than that in spermatogenic cells.The results hint that Crb3 not only relates with the establishment and maintenance of seminiferous epithelium cells, but also probably participates in regulating the synthesis and secretion of steroid hormone in testis.