兽类学报 ›› 2017, Vol. 37 ›› Issue (2): 189-199.DOI: 10.16829/j.slxb.201702010

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双峰驼睾丸和隐睾细胞外基质相关蛋白的组织化学特征及超微结构比较

袁莉刚 鲁玉荣 陶金忠 张勇   

  1. 甘肃农业大学动物医学院
  • 出版日期:2017-04-20 发布日期:2017-04-20

Comparison of histochemical and ultrastructural characteristics of extracellular matrix components in the scrotal and cryptorchid testes of the Bactrian camel

YUAN Ligang,LU Yurong,TAO Jinzhong,ZHANG Yong   

  1. College of Veterinary Medicine, Gansu Agricultural University
  • Online:2017-04-20 Published:2017-04-20

摘要: 为探索细胞外基质相关蛋白在隐睾双峰驼的分布情况及其组织化学特征,应用电镜技术和多种组织化学方法比较了隐睾和正常睾丸的超微结构,组织化学特点及层粘连蛋白(LN)、Ⅳ型胶原(Col Ⅳ)和硫酸乙酰肝素糖蛋白(HSPG)的分布特征。结果显示:(1)与正常睾丸间质结构相比,光镜下隐睾生精小管发育不全,间质内胶原纤维稀疏,网状纤维分布明显,间质血管及生精小管固有膜PAS及AB-PAS阳性反应较弱。电镜下,隐睾生精上皮基膜明显增生,外围I型胶原纤维较少,管周肌样细胞不典型;间质毛细血管及Leydig细胞周围纤维细胞多见,而正常睾丸在间质毛细血管及Leydig细胞周围多分布有成纤维细胞。(2) 免疫组织化学染色显示,正常睾丸组织的Col Ⅳ、LN及HSPG在Leydig细胞内均为强阳性表达,Col Ⅳ和LN在毛细血管内皮细胞强阳性表达,后者在Sertoli细胞的表达尤为明显,HSPG在精原细胞无表达;隐睾时Col Ⅳ、LN及HSPG在Leydig细胞内阳性表达均明显减弱,Col Ⅳ、LN在管周肌样细胞及毛细血管内皮细胞阳性表达也减弱明显,HSPG在精原细胞较强阳性表达,且在精子细胞呈强阳性表达。免疫组织化学图像分析结果显示,双峰驼正常睾丸组织中Col Ⅳ和LN的分布显著高于隐睾组织(P<0.05),HSPG检测结果在正常睾丸与隐睾之间无统计学差异(P>0.01)。该研究表明,双峰驼隐睾生精小管发育异常,间质组织中合成胶原纤维的能力下降,睾丸细胞外基质的重要成分Col Ⅳ,LN与正常组差异显著与生精小管及Leydig细胞异常发育有关,而HSPG在隐睾生精上皮的强阳性表达与精原细胞发育不成熟密切相关。

关键词: 双峰驼, 隐睾, 层粘连蛋白, 胶原蛋白, 硫酸乙酰肝素糖蛋白

Abstract:

he present study compared the histochemical and ultrastructural characteristics of the extracellular matrix components between the scrotal and bilateral cryptorchid testes of same-aged Bactrian camels. The scrotal and cryptorchid testes of 2-year-old Bactrian camel were prepared for light and electron microscopy using histochemistry and transmission electron microscopy methods. Image-Pro Plus (IPP) statistical methods were used to identify the characteristic indices for laminin (LN), type IV collagen (Col IV), and heparan sulfate proteoglycan (HSPG) distribution by immunohistochemistry methods. The observations made with light microscopy showed that, compared with the scrotal testes, the seminiferous tubules in the bilateral cryptorchid of Bactrian camels were underdeveloped and the collagen fibers were sparse, but the reticular fibers were distributed in interstitial tissues. The PAS-positive and AB-PAS reactions in the cryptorchidism basement membranes and capillaries were weakly stained. Electron micrographs of the seminiferous tubules showed that the fibrocytes were always located around the cryptorchid interstitial blood capillaries and Leydig cells, but instead of fibroblasts and collagen in the interstitial tissues of the scrotal testes, the seminiferous epithelial basement membranes were hyperplasic surrounding spare peripheral type I collagen fibers and the peritubular myoid cells were atypical. Immunostaining analysis showed that the Col IV, LN, and HSPG were all present in Leydig cells, and Col IV and LN were strongly expressed in the walls of the small vessels. The LN were strongly expressed in Sertoli cells, but HSPG was not expressed in the spermatogonia of the Bactrian camel scrotal testes. In the cryptorchidism testes, the expression of Col IV, LN, and HSPG were weakly expressed in Leydig cells,and Col IV and LN were significantly reduced in the endothelial cells of the blood capillaries and peritubular myoid cells, but HSPG was expressed more strongly in the spermatogonia and markedly present in the spermatids. The average absorbance of Col IV and LN expression was significantly higher in the Bactrian camel scrotal testes (P<0.05). In addition, the average absorbance of HSPG expression did not differ between the healthy and cryptorchid Bactrian camels (P>0.01). Taken together, the abnormalities in the seminiferous tubule and the decrease in the synthetic ability of interstitial collagen fibers are the histochemical and ultrastructural characteristics of cryptorchid Bactrian camels. The average absorbances of Col IV and LN expression were significantly lower in the cryptorchid testes were correlated with the developmental abnormalities of seminiferous tubules and Leydig cells, and the expression of HSPG was significant in the seminiferous epithelium and closely related to spermatogonia development.
 

Key words: Camelus bactrianus, Cryptorchidism, Heparan sulfate proteoglycans, CollagenⅣ, Laminin