不同月龄高原型藏绵羊睾丸组织中蛋白基因产物9.5和神经肽Y的分布比较

Distribution of Protein Gene Product 9.5 and Neuropeptide Y in Tibetan Plateau Sheep Testis at Different Ages

旨在比较蛋白基因产物9.5(PGP9.5)和神经肽Y(NPY)在不同发育时期藏绵羊睾丸的组织分布特征,探索其对藏绵羊睾丸发育及生殖机能调节的相关作用。采用睾丸摘除手术收集0.5、2、5、8、12及18月龄藏绵羊睾丸,应用免疫组织化学SP法及IPP图像分析技术研究PGP9.5和NPY的分布特征。PGP9.5和NPY在0.5月龄藏绵羊睾丸组织表达量较低,生殖母细胞中基本无表达,但2月龄时,PGP9.5表达量增加极显著(P<0.01),且同年龄段PGP9.5表达量均极显著高于NPY(P<0.01),至18月龄时,PGP9.5和NPY表达量明显升高,且与0.5月龄相比差异极显著(P<0.01),二者在12月龄及以后初级精母细胞中表达均为阴性。PGP9.5在Sertoli细胞阳性信号随睾丸发育随年龄增加逐渐降低,至18月龄时基本无表达,其在各年龄段Leydig细胞及其周围血管壁呈阳性表达。NPY在Leydig细胞的阳性信号较弱,2月龄及以后各发育阶段睾丸Sertoli细胞中均为阳性表达,在血管壁的表达亦随年龄增加而增强。高原缺氧环境中,不同月龄高原型藏绵羊睾丸组织中PGP9.5和NPY的表达差异与Sertoli细胞发育及精子发生调控关系密切;PGP9.5在Leydig细胞强阳性,表明其与激素合成分泌相关,NPY表达随着月龄而增强,有利于加强睾丸血管舒缩以适应对低氧的调节,或在局部血液循环与Sertoli细胞之间物质传递发挥作用。

This experiment was conducted to investigate the distribution of gene product 9.5（PGP 9.5）and neuropeptide Y（NPY）in Tibetan Plateau sheep testis and in an attempt to explore their relationships with testicular spermatogenesis at different ages.Testis from 0.5-,2-,5-,8-,12-and 18-month-old Tibetan Plateau sheep were sectioned and immunostained with antisera to the NPY and PGP 9.5.Furthermore,IPP（Image-Pro Plus）statistics methods were used to compare the average absorbance of PGP9.5and NPY expression in all of these testis.The testicular parenchyma did not show any NPY-and PGP9.5-containing immunoreactivity in 0.5-month-old sheep,and very scattered immunoreactivities were present within the gonocytes,but the expression of PGP9.5in testis of 2-month-old sheep was significantly stronger than NPY at the same age（P〈0.01）,both of the PGP9.5and NPY were strongly present in 18-month-old sheep（P〈0.01）.By contrast,no expression of NPY and PGP9.5immunoreactivities were observed withinthe primary spermatocytes in 18-month-old sheep.The expression of PGP9.5in Sertoli cells underlie age-related declines,which was poorly investigated at 12-month-old and no expression at 18-month-old,but they were obviously detected in Leydig cells and vscellum at different ages.The NPY expression in Leydig cells were also poorly investigated,and were mainly expressed in Sertoli cells in 2-month-old sheep testis,and after that,the expression of NPY in vscellum with a manner of age-related enhances.The results suggest that the PGP9.5and NPY expression was significantly difference in Tibetan Plateau sheep testis at different ages might serve as regulators for the testicular functions,also the expression of PGP9.5in Leydig cells in the testis reflect the functional regulates of testosterone.Namely,the distribution of NPY in the wall of testis vascular significantly stronger in a manner of age-related increase may provide new indexes for the research of plateau adaptability mechanism in testicular blood regulation,and play a role in the local microcirculation and the interactions with the Sertoli cells.