不同发育阶段关中奶山羊睾丸间质细胞的生物学特性  

Biological Characteristics of Leydig Cells in Different Developmental Stages of Guanzhong Dairy Goats

作　　者：武永杰[1] 范宁娟[2] 陈鸿 赵芳娥 马琳[1] 徐永平[1] WU Yongjie;FAN Ningjuan;CHEN Hong;ZHAO Fang’e;MA Lin;XU Yongping(College of Veterinary Medicine,Northwest Agriculture and Forestry University,Yangling 712100,China;College of Life Sciences,Northwest Agriculture and Forestry University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《云南农业大学学报（自然科学版）》2024年第5期31-39,共9页Journal of Yunnan Agricultural University:Natural Science

基　　金：国家自然科学基金项目(32372969,32002242)。

摘　　要：【目的】研究幼龄期、初情期和成年期关中奶山羊睾丸间质细胞(Leydig cells,LCs)的形态、分布特点、超微结构特征及睾酮合成水平,解析不同发育阶段LCs的形态特征与睾酮合成水平之间的相关性。【方法】使用光镜和改良后的透射电镜技术观察不同月龄奶山羊LCs的形态、分布规律及超微结构特征;采用免疫组织化学染色方法检测睾酮合成关键酶3β-羟基类固醇脱氢酶的表达和分布;采用酶联免疫吸附试验检测血清中促黄体生成素(luteinizing hormone,LH)和睾酮含量的变化;采用荧光定量PCR和免疫印迹技术分析LCs成熟和睾酮合成关键基因/蛋白表达的规律。【结果】与幼龄期相比,初情期和成年期的LCs逐渐变大、呈不规则多边形,且逐渐向间质中央移动。初情期和成年期的LCs胞质内含有大量脂滴,且脂滴与线粒体、光面内质网等细胞器密切接触。随着性成熟,血清中的LH和睾酮含量显著或极显著增加,睾丸组织中与睾酮合成相关的基因表达水平显著或极显著上调。【结论】关中奶山羊LCs的睾酮合成能力与其发育成熟过程密切相关,初情期和成年期的LCs胞质内存在大量脂滴且与线粒体直接接触,这为形态学方法研究脂滴参与睾酮的合成提供了科学依据。[Purpose]To study the morphology,distribution characteristics,ultrastructure characteristics and testosterone synthesis level of Leydig cells(LCs)in Guanzhong dairy goats at juvenile stage,puberty,and adult stage.[Methods]The morphology,distribution and ultrastructural characteristics of LCs in dairy goats at different ages were observed by light microscope and modified transmission electron microscopy methods.Immunohistochemistry method was employed to detect the expression and distribution of 3 beta-hydroxysteroid dehydrogenase,which was the key enzymes of testosterone synthesis.The contents of luteinizing hormone(LH)and testosterone in serum were tested by enzyme-linked immunosorbent assay.The expression levels of LCs maturation-related genes and testosterone synthesis-related genes or proteins were analyzed by qPCR and Western-blotting.[Results]Compared with the juvenile stage,LCs in the puberty and adult stage gradually increased in size,formed irregular shapes,and gradually moved towards the center of the testicular interstitial.The cytoplasm of LCs in puberty and adult stage contained a large amount of lipid droplets,which were in close contact with organelles such as mitochondria and smooth endoplasmic reticulum.With the maturation of dairy goats,the contents of LH and testosterone in serum increased significantly or extremely significantly,and the expression levels of genes related to testosterone synthesis in testicular tissues were significantly or extremely significantly up-regulated.[Conclusion]The testosterone synthesis ability of Guanzhong dairy goat LCs is closely related to its development and maturation.There are a lot of lipid droplets in the cytoplasm of LCs during puberty and adult stage,which directly contact with mitochondria.The results provide a scientific basis for morphological study of lipid droplets involved in testosterone synthesis.

关 键 词：奶山羊 形态学方法 睾丸间质细胞 睾酮 脂滴 

分 类 号：S827.941[农业科学—畜牧学]