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目的探索细胞外基质相关蛋白在高原地区藏绵羊与小尾寒羊睾丸的分布及组织化学特征。方法应用组织化学方法、Image-Pro Plus(IPP)图像分析技术及电子显微镜,观察比较藏绵羊(4只)与小尾寒羊(5只)睾丸组织化学特点及层黏连蛋白(LN)、Ⅳ型胶原(ColⅣ)和硫酸乙酰肝素糖蛋白(HSPG)的分布特征。结果与小尾寒羊睾丸相比,藏绵羊生精小管基膜及间质组织内胶原纤维及网状纤维丰富;过碘酸-雪夫(PAS)及阿利新蓝-过碘酸-雪夫(AB-PAS)染色显示,藏绵羊睾丸间质血管及生精小管固有膜阳性反应更为丰富。免疫组织化学显示,ColⅣ在藏绵羊及小尾寒羊生精小管上皮均呈阳性表达,LN在藏绵羊Sertoli细胞及管周肌样细胞呈弱阳性表达,而在小尾寒羊Sertoli细胞、Leydig细胞及管周肌样细胞为中等阳性表达;HSPG在藏绵羊及小尾寒羊肌样细胞均呈强阳性表达,而在Sertoli细胞及Leydig细胞均为弱表达。免疫组织化学图像分析结果显示,藏绵羊睾丸组织中ColⅣ和LN的分布显著低于小尾寒羊睾丸组织(P<0.01),HSPG检测结果则显著高于小尾寒羊睾丸组织(P<0.01)。电子显微镜下观察,藏绵羊及小尾寒羊生精小管固有膜可见发育良好的生殖上皮基膜以及1层明显的Ⅰ型胶原纤维,藏绵羊固有膜胶原纤维层丰富且Leydig细胞内脂滴明显。结论高原环境下藏绵羊睾丸固有膜及间质结缔组织较为丰富,在一定程度上影响了生精上皮发育;藏绵羊睾丸间质血管壁及生精小管固有膜AB-PAS阳性反应增强与Leydig细胞分泌功能相关,小尾寒羊睾丸组织LN表达显著增加及HSPG显著降低与生精上皮发育程度关系密切。
Objective To explore the distribution and histochemical characteristics of extracellular matrix-associated proteins in Tibetan sheep and Small Tail Han sheep in the plateau. Methods The histochemical characteristics and laminin (LN), Ⅳ (Ⅳ) mRNA expression in testis of Tibetan sheep (4) and Small Tail Han sheep (5) were observed by histochemical method and Image- Pro Plus (IPP) Col Ⅳ and Heparan Sulfate Glycoprotein (HSPG) distribution characteristics. Results Compared with the testis in the Small Tail Han sheep, the collagen fibers and reticular fibers were abundant in the seminiferous tubules and interstitium of Tibetan sheep. PAS) staining showed that Tibetan sheep testicular interstitial blood vessels and seminiferous tubules inherent membrane positive reaction is more abundant. Immunohistochemistry showed that ColⅣ was positive in seminiferous tubules of Tibetan sheep and Small Tail Han sheep, while LN was weakly positive in Sertoli cells and perivascular myoid cells of Tibetan sheep, while in Sertoli cells, Leydig cells and Tubular myoid-like cells were moderately positive expression; HSPG in Tibetan sheep and Small Tail sheep muscle-like cells were strongly positive expression, but in Sertoli cells and Leydig cells were weakly expressed. Immunohistochemical image analysis showed that the distribution of ColⅣ and LN in testis tissue of Tibetan sheep was significantly lower than that in Small Tail Han sheep (P <0.01), but HSPG was significantly higher than that in Small Tail Han sheep (P <0.01). Under the electron microscope, the well-developed genital epithelial basal lamina and type 1 collagen fibers were observed in the lamina propria of Tibetan sheep and Small Tail Han sheep. The lamina propria of Tibetan Sheep was abundant and the lipid droplets in Leydig cells were obvious. Conclusion The testicular lamina propria and interstitial connective tissue of Tibetan sheep were abundant in the plateau environment, which affected the development of the seminiferous epithelium to a certain extent. The positive reaction of AB-PAS in the testicular stromal wall and the lamina propria of Tibetan sheep was enhanced with that of Leydig cells Secretion, LN in testis tissue of small-tailed Han sheep significantly increased and HSPG decreased significantly with the development of seminiferous epithelial cells closely related.