香猪睾丸间质细胞TAS1R3基因干扰后对自噬相关因子的影响

doi:10.11843/j.issn.0366-6964.2023.04.016

Abstract

Abstract: The purpose of this study was to explore the effect of TAS1R3 gene on the expression of autophagy-related genes in Congjiang Xiang pigs through mTOR. In this experiment, three 30-day-old healthy male Congjiang Xiang pigs were selected, testis tissue was collected and Leydig cells were cultured. The interference vector and empty vector were transfected into the cells, and their effects on the expression of TAS1R3 gene were detected by qRT-PCR. The effects of TAS1R3 gene interference on the expression of taste receptor family 1 receptor (TAS1R1), extracellular signal-regulated kinase 1 (ERK1), extracellular signal-regulated kinase 2 (ERK2), rapamycin target protein (mTOR), BECN1 and microtubule-associated protein 1 light chain 3 β (MAP1LC3B) were detected by qRT-PCR and Western blot. The results showed that when TAS1R3 gene was interfered, compared with shRNA-NC control group, the mRNA expression of TAS1R1, ERK1, ERK2 and mTOR genes decreased significantly(P<0.01), while the expression of autophagy factors BECN1 and MAP1LC3B increased significantly (P<0.01). The results of Western blot showed that when TAS1R3 gene was silenced, compared with shRNA-NC control group, the expression of T1R3, T1R1, p-mTOR, p-S6K1 and p-ERK12 decreased significantly(P<0.01), while the expression of autophagy protein Beclin 1 increased significantly(P<0.01). In this experiment, by transfecting TAS1R3 gene interference vector into Leydig cells, it was found that when TAS1R3 gene was interfered, it could affect the expression of mTOR gene, and then regulate the expression of downstream autophagy factors, suggesting that TAS1R3 gene is closely related to autophagy of Leydig cells, which provides a reference for further study on the regulation mechanism of TAS1R3 gene affecting the reproduction of Xiang pigs by regulating autophagy.

Key words: Congjiang Xiang pig, T1R3, RNA interference, leydig cell, autophagy

CLC Number:

S828.2

WANG Han, MENG Lijie, LIU Wenjiao, XU Yongjian, GONG Ting. Effect of TAS1R3 Gene Interference on Autophagy Related Factors in Leydig Cells of Xiang Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1525-1534.

References 35

[1]	TEMUSSI P A.Sweet, bitter and umami receptors:A complex relationship[J].Trends Biochem Sci, 2009, 34(6):296-302.
[2]	YARMOLINSKY D A, ZUKER C S, RYBA N J P.Common sense about taste:From mammals to insects[J].Cell, 2009, 139(2):234-244.
[3]	SIMON S A, ROPER S D.Morphology of vertebrate taste organs and their nerve supply[J]. Mechanisms of Taste Transduction, 1993:29-82.
[4]	YOSHIDA R, NINOMIYA Y.Taste information derived from T1R-expressing taste cells in mice[J]. Biochem J, 2016, 473(5):525-536.
[5]	NUEMKET N, YASUI N, KUSAKABE Y, et al.Structural basis for perception of diverse chemical substances by T1r taste receptors[J]. Nat Commun, 2017, 8:15530.
[6]	MUROVETS V O, SOZONTOV E A, ZACHEPILO T G.The effect of the taste receptor protein T1R3 on the development of islet tissue of the murine pancreas[J].Dokl Biol Sci, 2019, 484(1):1-4.
[7]	WU B, ELDEGHAIDY S, AYED C, et al.Mechanisms of umami taste perception:from molecular level to brain imaging[J].Crit Rev Food Sci Nutr, 2022, 62(25):7015-7024.
[8]	XIE S Z, YANG G, JIANG X M, et al.Polygonatum cyrtonema Hua Polysaccharide promotes GLP-1 secretion from enteroendocrine L-Cells through sweet taste receptor-mediated cAMP signaling[J].J Agric Food Chem, 2020, 68(25):6864-6872.
[9]	HASS N, SCHWARZENBACHER K, BREER H.T1R3 is expressed in brush cells and ghrelin-producing cells of murine stomach[J].Cell Tissue Res, 2010, 339(3):493-504.
[10]	龚婷.甜味受体T1R3与Gα-味导素在小鼠睾丸类固醇激素合成中的功能及作用机制研究[D].南京:南京农业大学, 2016.GONG T.Roles of sweet taste receptor T1R3 and Alpha-gustducin in testicular streroidogenesis of mice[D].Nanjing:Nanjing Agricultural University, 2016.(in Chinese)
[11]	GONG T, WANG W Y, XU H Q, et al.Longitudinal expression of testicular TAS1R3 from prepuberty to sexual maturity in Congjiang Xiang Pigs[J].Animals, 2021, 11(2):437.
[12]	LEE A A, OWYANG C.Sugars, Sweet taste receptors, and brain responses[J].Nutrients, 2017, 9(7):653.
[13]	SAXTON R A, SABATINI D M.mTOR signaling in growth, metabolism, and disease[J].Cell, 2017, 168(6):960-976.
[14]	KIM Y C, GUAN K L.mTOR:A pharmacologic target for autophagy regulation[J].J Clin Invest, 2015, 125(1):25-32.
[15]	ZHOU Y F, REN J, SONG T X, et al.Methionine regulates mTORC1 via the T1R1/T1R3-PLCβ-Ca2+-ERK1/2 signal transduction process in C2C12 cells[J].Int J Mol Sci, 2016, 17(10):1684.
[16]	WAUSON E M, ZAGANJOR E, COBB M H.Amino acid regulation of autophagy through the GPCR TAS1R1-TAS1R3[J].Autophagy, 2013, 9(3):418-419.
[17]	WAUSON E M, ZAGANJOR E, LEE A Y, et al.The G protein-coupled taste receptor T1R1/T1R3 regulates mTORC1 and autophagy[J].Mol Cell, 2012, 47(6):851-862.
[18]	LIU J Q, WANG Y H, LI D W, et al.Milk protein synthesis is regulated by T1R1/T1R3, a G protein-coupled taste receptor, through the mTOR pathway in the mouse mammary gland[J].Mol Nutr Food Res, 2017, 61(9):1601017.
[19]	朱宇辰.氟对小鼠睾丸组织及间质细胞自噬的体内外观察[D].晋中:山西农业大学, 2016.ZHU Y C.A research of fluoride on autophagy in mice testis and Leydig cells:in vivo and vitro[D].Jinzhong:Shanxi Agricultural University, 2016.(in Chinese)
[20]	赵祥龙.尼古丁通过睾丸间质细胞自噬降低血清睾酮分泌[D].上海:上海交通大学, 2018.ZHAO X L.Nicotine decreases serum testosterone via autophagy in TM3 cells[D].Shanghai:Shanghai Jiao Tong University, 2018.(in Chinese)
[21]	MEYER D, VOIGT A, WIDMAYER P, et al.Expression of Tas1 taste receptors in mammalian spermatozoa:functional role of Tas1r1 in regulating basal Ca2+ and cAMP concentrations in spermatozoa[J].PLoS One, 2012, 7(2):e32354.
[22]	GAO F Y, LI G P, LIU C, et al.Autophagy regulates testosterone synthesis by facilitating cholesterol uptake in Leydig cells[J].J Cell Biol, 2018, 217(6):2103-2119.
[23]	王维勇.睾丸T1R3参与从江香猪初情期睾酮合成的机制研究[D].贵阳:贵州大学, 2020.WANG W Y.Study on testicular T1R3 involved in testosterone synthesis of Congjiang Xiang Pig at puberty[D].Guiyang:Guizhou University, 2020.(in Chinese)
[24]	LUDDI A, GOVERNINI L, WILMSKÖTTER D, et al.Taste receptors:new players in sperm biology[J].Int J Mol Sci, 2019, 20(4):967.
[25]	NICKLIN P, BERGMAN P, ZHANG B L, et al.Bidirectional transport of amino acids regulates mTOR and autophagy[J].Cell, 2009, 136(3):521-534.
[26]	YAN X J, ZHOU R M, MA Z Y.Autophagy-cell survival and death[J]. Adv Exp Med Biol, 2019:667-696.
[27]	XU H D, QIN Z H.Beclin 1, Bcl-2 and autophagy[J]. Adv Exp Med Biol, 2019:109-126.
[28]	ZHENG L F, ZHANG W, ZHOU Y F, et al.Recent advances in understanding amino acid sensing mechanisms that regulate mTORC1[J].Int J Mol Sci, 2016, 17(10):1636.
[29]	张伟. 参与mTOR活性调控的G蛋白鉴定及猪源T1R1/T1R3感应氨基酸筛选和功能研究[D].武汉:华中农业大学, 2015.ZHANG W.Identification of G protein that participates in the regulation of mTOR signaling and screening of amino acids sensed by swine T1R1/T1R3 and study of the function[D].Wuhan:Huazhong Agricultural University, 2015.(in Chinese)
[30]	任娇. 成肌细胞中T1R1/T1R3介导调控mTOR信号的氨基酸筛选及分子机制[D].武汉:华中农业大学, 2015.REN J.Screening of amino acids participating in the regulation of M TOR signaling by T1R1/T1R3 and the mechanism in myoblast[D].Wuhan:Huazhong Agricultural University, 2015.(in Chinese)
[31]	MÄKELÄ J A, KOSKENNIEMI J J, VIRTANEN H E, et al.Testis development[J].Endocr Rev, 2018, 40(4):857-905.
[32]	PLANT T M, ZELEZNIK A J.Knobil and Neill's physiology of reproduction[M].4th ed.Amsterdam:Academic Press, 2015.
[33]	ZHU Y C, YIN Q Q, WEI D D, et al.Autophagy in male reproduction[J].Syst Biol Reprod Med, 2019, 65(4):265-272.
[34]	LV C Y, WANG X L, GUO Y, et al.Role of selective autophagy in spermatogenesis and male fertility[J].Cells, 2020, 9(11):2523.
[35]	CHEN X W, LI C, CHEN Y, et al.Aflatoxin B1 impairs leydig cells through inhibiting AMPK/mTOR-mediated autophagy flux pathway[J].Chemosphere, 2019, 233:261-272.

Effect of TAS1R3 Gene Interference on Autophagy Related Factors in Leydig Cells of Xiang Pig

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