Expression and Distribution of Autophagy-related Molecules During Epididymal Development in Congjiang Xiang Pig (Sus scrofa)
WANG Han1,2, MENG Li-Jie1,2, GONG Ting1,2,*
1 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; 2 College of Animal Science, Guizhou University, Guiyang 550025, China
Abstract：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-Ⅰ was 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 might be an autophagy regulation pathway through mTOR in the epididymis of Congjiang Xiang pigs at different ages. This study provides basic data for exploring the regulatory mechanism of autophagy in epididymis development.
王涵, 蒙利洁, 龚婷. 从江香猪附睾发育中自噬相关分子的表达与分布[J]. 农业生物技术学报, 2022, 30(12): 2354-2362.
WANG Han, MENG Li-Jie, GONG Ting. Expression and Distribution of Autophagy-related Molecules During Epididymal Development in Congjiang Xiang Pig (Sus scrofa). 农业生物技术学报, 2022, 30(12): 2354-2362.
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