|
|
|
| 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.
|
|
Received: 12 January 2022
|
|
|
|
Corresponding Authors:
* tgong@gzu.edu.cn
|
|
|
|
[1] 蒙利洁, 王维勇, 徐永健, 等. 2021. 从江香猪AR在附睾组织发育过程中的表达模式[J]. 农业生物技术学报, 29(6): 1103-1112.
(Meng L J, Wang W Y, Xu Y J, et al.Expression pattern of AR in Congjiang Xiang pig during epididymal tissue development[J]. Journal of Agricultural Biotechnology, 29(6): 1103-1112.)
[2] 蒙利洁, 王维勇, 杨艺, 等. 2020. T1R1和T1R3在从江香猪附睾发育中的表达模式[J]. 畜牧兽医学报, 51(11): 2720-2730.
(Meng L J, Wang W Y, Yang Y, et al.2020. Expression patterns of T1R1 and T1R3 in epididymis development of Congjiang Xiang pig[J]. Journal of Animal Husbandry and Veterinary Medicine, 51(11): 2720-2730.)
[3] 张友旺, 刘俊, 马开槐, 等. 2019. 自噬对精子发生及活力的影响[J]. 中华男科学杂志, 25(01): 72-76.
(Zhang Y W, Liu J, Ma K H, et al.2019. Effect of autophagy on spermatogenesis and motility[J]. Chinese Journal of Andrology, 25(01): 72-76.)
[4] 朱文倩, 张学明. 2021. 附睾结构及其功能研究进展[J]. 中国兽医学报, 41(6): 1195-1209.
(Zhu W Q, Zhang X M.2021. Research progress on structure and function of epididymis[J]. Chinese Journal of Veterinary Medicine, 41(6): 1195-1209.)
[5] Abdel-Maksoud F M, Hussein M T, Attaai A.2019. Seasonal variation of the intraepithelial gland in camel epididymis with special reference to autophagosome[J]. Microscopy and Microanalysis, 25(4): 1052-1060.
[6] Baechler B L, Bloemberg D, Quadrilatero J.2019. Mitophagy regulates mitochondrial network signaling, oxidative stress, and apoptosis during myoblast differentiation[J]. Autophagy, 15(9): 1606-1619.
[7] Busada J T, Chappell V A, Niedenberger B A, et al.2015. Retinoic acid regulates Kit translation during spermatogonial differentiation in the mouse[J]. Developmental Biology, 397(1): 140-149.
[8] Cheng C, Liu Z G.2019. Autophagy and the metabolism of misfolding protein[J]. Advances in Experimental Medicine and Biology, 1206: 375-420.
[9] Dacheux J L, Dacheux F.2014. New insights into epididymal function in relation to sperm maturation[J]. Reproduction, 147(2): R27-R42.
[10] De Santi F, Lunardi I D, Beltrame F L, et al.2020. Muscular atrophy, impaired epithelial autophagy and UCHL1 increase in androgen-deficient cauda epididymis[J]. Reproduction, 159(6): 693-705.
[11] Fan X, Liu Y, Yue M, et al.2021. Effect of cryptorchidism on the histomorphometry, proliferation, apoptosis, and autophagy in boar testes[J]. Animals, 11(5): 1379.
[12] Galluzzi L, Green D.2019. Autophagy-independent functions of the autophagy machinery[J]. Cell, 177(7): 1682-1699.
[13] Gao H, Khawar M B, Li W.2019. Autophagy in reproduction[J]. Advances in Experimental Medicine and Biology, 1206: 453-468.
[14] Gao F Y, Li J P, Liu C, et al.2018. Autophagy regulates testosterone synthesis by facilitating cholesterol uptake in leydig cells[J]. The Journal of Cell Biology, 217(6): 2103-2119.
[15] Gervasi M G, Visconti P E.2017. Molecular changes and signaling events occurring in spermatozoa during epididymal maturation[J]. Andrology, 5(2): 204-218.
[16] Hga B, Yo A, Hy A, et al.2021.Induction of autophagy via the ROS-dependent AMPK-mTOR pathway protects copper-induced spermatogenesis disorder[J]. Redox Biology, 49: 102227.
[17] Huang Q, Liu Y, Zhang S, et al.2021. Autophagy core protein atg5 is required for elongating spermatid development, sperm individualization and normal fertility in male mice[J]. Autophagy, 17(7): 1753-1767.
[18] James E R, Carrell D T, Aston K I, et al.2020. The role of the epididymis and the contribution of epididymosomes to mammalian reproduction[J]. International Journal of Molecular Sciences, 21(15): 5537.
[19] Kalachev A V, Yurchenko O V, Kiselev K V.2019. Macroautophagy is involved in residual bodies formation during spermatogenesis in sea urchins, Strongylocentrotus intermedius[J]. Tissue and Cell, 56: 79-82.
[20] Levine B, Kroemer G.2008. Autophagy in the pathogenesis of disease[J]. Cell, 132(1): 27-42.
[21] Levine B, Kroemer G.2019. Biological functions of autophagy genes: A disease perspective[J]. Cell, 176(1-2): 11-42.
[22] Li W, Zhang L.2019a. Regulation of ATG and autophagy initiation[J]. Advances in Experimental Medicine and Biology, 1206: 41-65.
[23] Li Y J, Chen Y Y.2019b. AMPK and autophagy[J]. Advances in Experimental Medicine and Biology, 1206: 85-108.
[24] Lin P W, Chu M L, Liu H S.2020. Autophagy and metabolism[J], The Kaohsiung Journal of Medical Sciences, 37(1): 12-19.
[25] Lu J, Wu M, Yue Z.2020. Autophagy and Parkinson's disease[J]. Advances in Experimental Medicine and Biology, 1207: 21-51.
[26] Meng L J, Wang W Y, Xu Y J, et al.2020. Postnatal differentiation and regional histological variations in the ductus epididymidis of the Congjiang Xiang pig[J]. Tissue and Cell, 67: 101411.
[27] Qi Z, Chen L.2019. Endoplasmic reticulum stress and autophagy[J]. Advances in Experimental Medicine and Biology, 1206: 167-177.
[28] Sahin P, Gungor-Ordueri N, Celik-Ozenci C.2018. Inhibition of mTOR pathway decreases the expression of pre-meiotic and meiotic markers throughout postnatal development and in adult testes in mice[J]. Andrologia, 50: e12811.
[29] Schimming B, Baumam C, Pinheiro P, et al.2017. Aquaporin 9 is expressed in the epididymis of immature and mature pigs[J]. Reproduction in Domestic Animals, 52: 617-624.
[30] Tabecka-Lonczynska A, Mytych J, Solek P, et al.2020. Autophagy as a consequence of seasonal functions of testis and epididymis in adult male European bison (Bison bonasus, Linnaeus 1758)[J]. Cell and Tissue Research, 379(3): 613-624.
[31] Tanida I, Ueno T, Kominami E.2008. LC3 and autophagy[J]. Methods in Molecular Biology, 445: 77-88.
[32] Tarique I, Tariq M, Bai X, et al.2020. Interaction of epididymal epithelia and their secretions with spermatozoa supports functional and morphological changes during long-term storage in the Chinese soft-shelled turtle (Pelodiscus sinensis)[J]. Microscopy and Microanalysis, 26(3): 542-550.
[33] Torres-Quiroz F, Filteau M, Landry C R.2015. Feedback regulation between autophagy and PKA[J]. Autophagy, 11(7): 1181-1183.
[34] Wang Y, Zhang H.2019. Regulation of autophagy by mTOR signaling pathway[J]. Advances in Experimental Medicine and Biology, 1206: 67-83.
[35] Wei Z, De I, Dun M, et al.2018. Characteristics of the epididymal luminal environment responsible for sperm maturation and storage[J]. Frontiers in Endocrinology, 9: 59.
[36] Wong S Q, Kumar A V, Mills J, et al.2020. Autophagy in aging and longevity[J]. Human Genetics, 139(3): 277-290.
[37] Xi H, Ren F, Li Y, et al.2021. Changes in histology, protein expression, and autophagy in dairy goat testes during non-breeding season[J]. Biology of Reproduction, 105(5): 5.
[38] Xu H, Qin Z.2019. Beclin1, Bcl-2 and autophagy[J]. Advances in Experimental Medicine and Biology, 1206: 109-126.
[39] Yan X J, Zhou R, Ma Z Y.2019. Autophagy-cell survival and death[J]. Advances in Experimental Medicine and Biology, 1206: 667-696.
[40] Yang J, Zhou R, Ma Z.2019. Autophagy and energy metabolism[J]. Advances in Experimental Medicine and Biology, 1206: 329-357.
[41] Zhou W, Stanger S J, Anderson A L, et al.2019. Mechanisms of tethering and cargo transfer during epididymosome-sperm interactions[J]. BMC Biology, 17(1): 1-18. |
|
|
|
