从江香猪附睾发育中自噬相关分子的表达与分布  

Expression and Distribution of Autophagy-related Molecules During Epididymal Development in Congjiang Xiang Pig(Sus scrofa)

作　　者：王涵 蒙利洁 龚婷[1,2] WANG Han;MENG Li-Jie;GONG Ting(Key Laboratory of Plateau Mountain Animal Genetics,Breeding and Reproduction,Ministry of Education/Key Laboratory of Animal Genetics,Breeding and Reproduction,Guizhou Province,Guizhou University,Guiyang 550025,China;College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室/贵州省动物遗传育种与繁殖重点实验室,贵阳550025 [2]贵州大学动物科学学院,贵阳550025

出　　处：《农业生物技术学报》2022年第12期2354-2362,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31702117);贵州省科学技术项目(黔科合基础[2020]1Y135,黔科合基础[2018]1404)。

摘　　要：自噬作为一种主要的降解/再循环途径,在维持细胞内稳态和生殖发育中发挥了重要作用。为探讨自噬在公猪(Sus scrofa)附睾发育过程中发挥的潜在作用,本研究以从江香猪附睾为对象,利用Western blot、免疫组织化学染色和qRT-PCR方法,分析了附睾在出生后15 d(初情期前)、30 d(初情期)、60 d(初情期后)和180 d(性成熟期)4个时期自噬相关分子的表达和分布。Western blot结果显示,Beclin1在60 d表达量最高、180 d表达量最低;哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)在30 d表达量最高、180 d最低;微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,MAP1LC3/LC3)-Ⅱ/LC3-Ⅰ在180 d最高,在30和60 d最低。免疫组织化学染色结果显示,Beclin1在15、30和180 d集中分布于附睾上皮主细胞和微绒毛,在60 d主要分布于附睾上皮周围的肌样细胞;LC3在15、30和60 d附睾中集中分布于主细胞和微绒毛,但180 d在上皮窄细胞有特异性表达。qRT-PCR结果显示,自噬相关基因Beclin1表达量在15 d最高、180 d最低,15和30 d差异不显著;MAP1LC3B基因表达量在180 d最高、60 d最低,各日龄段差异显著(P<0.05)。自噬相关蛋白12(autophagy related protein 12,Atg12)和Unc-51样激酶1(Unc-51-like kinase 1,ULK1)基因表达量变化趋势一致,在180 d最高、60 d最低,各日龄段差异显著(P<0.05);mTOR基因表达量在30 d最高、15 d最低,各日龄段差异显著(P<0.05)。上述结果提示,从江香猪附睾发育过程中可能存在mTOR介导的自噬调节通路。本研究为探讨自噬参与附睾发育的调节机制提供基础资料。As a major degradation/recycling pathway,autophagy plays an important role in maintaining intracellular homeostasis and reproductive development.In order to explore the potential role of autophagy in the development of epididymis in boars(Sus scrofa),the expression and distribution of autophagy-related molecules in the epididymis of Congjiang Xiang pig were analyzed by Western blot,immunohistochemistry,and qRT-PCR methods at 15 d(pre-puberty),30 d(in-puberty),60 d(post-puberty)and 180 d(sexual maturity)after birth.The results of Western blot showed that the expression of Beclin1 was the highest at 60 d and the lowest at 180 d,the expression of mammalian target of rapamycin(mTOR)was the highest at 30 d and the lowest at 180 d,and the expression of microtubule-associated protein 1 light chain 3(MAP1LC3/LC3)-Ⅱ/LC3-Ⅰwas the highest at 180 d and the lowest at 30 d.Immunohistochemical staining results showed that Beclin1 protein was mainly distributed in the main cells and microvilli of epididymal epithelium at 15,30,and 180 d,while it was strongly positive in the muscle-like cells around the epididymal epithelium at 60 d.LC3 distributed mainly in the main cells and microvilli in the epididymis at 15,30,and 60 d,but was specifically expressed in the epithelial narrow cells at 180 d.The results of qRT-PCR showed that the expression of autophagy-related gene Beclin1 was the highest at 15 d,the lowest at 180 d,and there was no significant difference between 15 and 30 d;The expression of MAP1LC3B gene was the highest at 180 d and the lowest at 60 d,and there was a significant difference among different age groups(P<0.05).The expression of autophagy related protein 12(Atg12)and Unc-51-like kinase 1(ULK1)genes showed the same trend,with the highest at 180 d and the lowest at 60 d,with significant differences among different ages(P<0.05);The expression of mTOR gene was the highest at 30 d and the lowest at 15 d,and there was a significant difference among different age groups(P<0.05).Above results indicate that there migh

关 键 词：从江香猪 附睾 发育 自噬 

分 类 号：S828.2[农业科学—畜牧学]