YAP1在发情期不同繁殖力湖羊子宫内膜中的表达模式及功能分析

doi:10.11843/j.issn.0366-6964.2023.01.018

Abstract

Abstract: The aim of this study was to analyze the expression patterns and function of the effector gene YAP1 in Hippo signaling pathway in endometrium of Hu sheep with different fertility. Based on the genealogical archives and polymorphism analysis of BMPR-1B, a total of 9 healthy pluriparous ewes were divided into 3 groups (HBB, LBB and LB+, n=3). RNA was extracted from endometrial tissues of Hu sheep with different fertility, and reversed transcript into cDNA. Real-time quantitative PCR (RT-qPCR) was used to analyze the expression of several major genes in the Hippo signaling pathway in endometrium of Hu sheep with different fertility. The amino acid homology and genetic relationship of YAP1 protein in different species were analyzed by bioinformatics methods. Hu sheep endometrial stromal cells (ESCs) were isolated in vitro, and the expression of YAP1 was downregulated by using RNAi and cell transfection techniques in vitro. The effect of YAP1 gene on the expression of uterine receptivity related genes was analyzed by RT-qPCR. Moreover, the effect of interfering YAP1 on cell apoptosis was analyzed by flow cytometry, and the expression of apoptosis and mitochondria related genes was analyzed by RT-qPCR and Western blot. The expression levels of YAP1, LATS1 and MST1 in Hippo signaling pathway were different in endometrium tissues of Hu sheep with different fertility, and the expression level of YAP1 gene in high fertility Hu sheep (HBB) endometrium was significantly higher than that in low fertility Hu sheep (LB+, LBB) endometrium. Amino acid homology and phylogenetic tree analysis showed that YAP1 protein of goat and cattle had the highest homology and the closest genetic relationship with sheep. The mRNA expression levels of receptivity related genes (FOXO1, PRL, VEGF, OPN, COX-2) in ESCs were significantly changed after interfering YAP1. Meanwhile, the apoptosis rate of ESCs was significantly increased, the expression levels of Bax, BIM, p53, caspase3, caspase8, caspase9, the ratio of Bax/Bcl-2 were upregulated, and expression levels of Bcl-2 was downregulated after YAP1 silencing. In addition, the mRNA expression levels of genes related to mitochondrial function (FIS1, DNM1L, MFN2, PPARGC1A, OPA1) were also significantly changed. YAP1 may establish good endometrial receptivity by inhibiting apoptosis and stabilizing mitochondrial function.

Key words: Hu sheep, Hippo pathway, YAP1, uterine receptivity

CLC Number:

S826.3

WANG Yahan, ZHENG Yujing, ZHONG Pei, LI Xiaodan, WANG Feng. Expression Patterns and Functional Analysis of YAP1 in Endometrium of Hu Sheep with Different Fertility during Estrus[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 189-200.

References 36

[1]	张吉顺,刘念,谈姣嫣,等.湖羊消化道各段T1R1、T1R3及PepT1基因表达水平的分析[J].畜牧与兽医,2017,49(2):51-56.ZHANG J S,LIU N,TAN J Y,et al.Expression levels of T1R1/T1R3 and PepT1 genes in the gastrointestinal tract of Hu Sheep[J].Animal Husbandry & Veterinary Medicine,2017,49(2):51-56.(in Chinese)
[2]	OUYANG T H,MENG W,LI M H,et al.Recent advances of the Hippo/YAP signaling pathway in brain development and glioma[J].Cell Mol Neurobiol,2020,40(4):495-510.
[3]	李佳,周芳月,黄健,等.Hippo信号通路在雌性生殖系统中的研究进展[J].中国细胞生物学学报,2014,36(12):1689-1694.LI J,ZHOU F Y,HUANG J,et al.Research progress on Hippo signaling pathway in female reproductive system[J].Chinese Journal of Cell Biology,2014,36(12):1689-1694.(in Chinese)
[4]	AN S Y,GAO X X,WANG Z B,et al.Estradiol-17β regulates proliferation and apoptosis of sheep endometrial epithelial cells by regulating the relative abundance of YAP1[J].Anim Reprod Sci,2020,215:106328.
[5]	CHEN H X,SONG Y,YANG S Y,et al.YAP mediates human decidualization of the uterine endometrial stromal cells[J].Placenta, 2017,53:30-35.
[6]	吉新彦,钟国轩,赵斌.哺乳动物Hippo信号通路分子机制研究进展[J].遗传,2017,39(7):546-567.JI X Y,ZHONG G X,ZHAO B.Molecular mechanisms of the mammalian Hippo signaling pathway[J].Hereditas (Beijing),2017, 39(7):546-567.(in Chinese)
[7]	PICCOLO S,DUPONT S,CORDENONSI M.The biology of YAP/TAZ:Hippo signaling and beyond[J].Physiol Rev,2014,94(4):1287-1312.
[8]	ZHANG T,GUO S,ZHOU H,et al.Endometrial extracellular matrix rigidity and IFNτ ensure the establishment of early pregnancy through activation of YAP[J].Cell Prolif,2021,54(2):e12976.
[9]	许祺欣,苏仁伟.胚胎着床的过程与机制研究进展[J].生理学报,2020,72(1):91-104.XU Q X,SU R W.Research progress in process and mechanisms of embryo implantation[J].Acta Physiologica Sinica, 2020,72(1):91-104.(in Chinese)
[10]	OCHOA-BERNAL M A,FAZLEABAS A T.Physiologic events of embryo implantation and decidualization in human and non-human primates[J].Int J Mol Sci,2020,21(6):1973.
[11]	LI X D,YAO X L,XIE H Q,et al.PPP2R2A affects embryonic implantation by regulating the proliferation and apoptosis of Hu sheep endometrial stromal cells[J].Theriogenology,2021,176:149-162.
[12]	张涛.IFN-τ和生物力激活YAP维持奶牛生殖能力的作用及机制研究[D].武汉:华中农业大学,2021.ZHANG T.Study on the function and mechanism of IFN-τ and biomechanical activating YAP to maintain reproductive ability of dairy cows[D].Wuhan:Huazhong Agricultural University,2021.(in Chinese)
[13]	陈万方.OPN N-糖基化修饰对子宫内膜容受性建立的调控机制研究[D].大连:大连医科大学,2019.CHEN W F.Mutation of N-linked glycosylation in osteopontin affected the establishment of endometrial receptivity[D]. Dalian:Dalian Medical University,2019.(in Chinese)
[14]	SHEKIBI M,HENG S,NIE G Y.MicroRNAs in the regulation of endometrial receptivity for embryo implantation[J].Int J Mol Sci,2022,23(11):6210.
[15]	CHA J,SUN X F,DEY S K.Mechanisms of implantation:strategies for successful pregnancy[J].Nat Med,2012,18(12):1754-1767.
[16]	KIM Y H,CHOI J,YANG M J,et al.A MST1-FOXO1 cascade establishes endothelial tip cell polarity and facilitates sprouting angiogenesis[J].Nat Commun,2019,10(1):838.
[17]	REN B.FoxO1 transcriptional activities in VEGF expression and beyond:a key regulator in functional angiogenesis?[J].J Pathol,2018,245(3):255-257.
[18]	WANG X H,VALLS A F,SCHERMANN G,et al.YAP/TAZ orchestrate VEGF signaling during developmental angiogenesis[J]. Dev Cell,2017,42(5):462-478.E7.
[19]	SOLDI R,MITOLA S,STRASLY M,et al.Role of αvβ3 integrin in the activation of vascular endothelial growth factor receptor-2[J]. EMBO J,1999,18(4):882-892.
[20]	GUO X,YI H,LI T C,et al.Role of vascular endothelial growth factor (VEGF) in human embryo implantation:clinical implications[J]. Biomolecules,2021,11(2):253.
[21]	MENTER D G,SCHILSKY R L,DUBOIS R N.Cyclooxygenase-2 and cancer treatment:understanding the risk should be worth the reward[J].Clin Cancer Res,2010,16(5):1384-1390.
[22]	SIERRA J C,HOBBS S,CHATURVEDI R,et al.Induction of COX-2 expression by helicobacter pylori is mediated by activation of epidermal growth factor receptor in gastric epithelial cells[J].Am J Physiol Gastrointest Liver Physiol,2013,305(2):G196-G203.
[23]	AURIEMMA R S,DEL VECCHIO G,SCAIRATI R,et al.The interplay between prolactin and reproductive system:focus on uterine pathophysiology[J].Front Endocrinol,2020,11:594370.
[24]	MAN S M,KANNEGANTI T D.Converging roles of caspases in inflammasome activation,cell death and innate immunity[J]. Nat Rev Immunol,2016,16(1):7-21.
[25]	XIE Z L,KOYAMA T,SUZUKI J,et al.Coronary reperfusion following ischemia-Different expression of bcl-2 and bax proteins,and cardiomyocyte apoptosis[J].Jpn Heart J,2001,42(6):759-770.
[26]	姚云峰,王宝成,王俊.促凋亡蛋白Bim在肿瘤治疗中的研究进展[J].肿瘤学杂志,2019,25(3):247-251.YAO Y F,WANG B C,WANG J.Research progress on pro-apoptotic protein Bim in tumor therapy[J].Journal of Chinese Oncology,2019,25(3):247-251.(in Chinese)
[27]	KIM Y S,LI X F,KANG K H,et al.Stigmasterol isolated from marine microalgae navicula incerta induces apoptosis in human hepatoma HepG2 cells[J].BMB Rep,2014,47(8):433-438.
[28]	ESKES R,DESAGHER S,ANTONSSON B,et al.Bid induces the oligomerization and insertion of Bax into the outer mitochondrial membrane[J].Mol Cell Biol,2000,20(3):929-935.
[29]	CHI X K,NGUYEN D,PEMBERTON J M,et al.The carboxyl-terminal sequence of bim enables bax activation and killing of unprimed cells[J].Elife,2020,9:e44525.
[30]	李书萍,李波,张阳.抑制YAP对宫颈癌细胞增殖、凋亡、迁移及p53、Fra-1表达的影响[J].遵义医科大学学报,2022, 45(1):31-36.LI S P,LI B,ZHANG Y.Inhibition of YAP on proliferation,apoptosis,migration and expression of p53 and FRA-1 in cervical cancer cells[J].Journal of Zunyi Medical University,2022,45(1):31-36.(in Chinese)
[31]	张舒文.CA3靶向抑制YAP1诱导肝癌HepG2细胞凋亡及机制研究[D].锦州:锦州医科大学,2020.ZHANG S W.Targeted inhibition of YAP1-induced apoptosis of hepatocelular carcinoma hepG2 cells by CA3 and its mechanism[D].Jinzhou:Jinzhou Medical University,2020.(in Chinese)
[32]	YU R,LENDAHL U,NISTÉR M,et al.Regulation of mammalian mitochondrial dynamics:opportunities and challenges[J].Front Endocrinol (Lausanne),2020,11:374.
[33]	肖成,胡进,纪红军,等.小鼠子宫内膜基质细胞中HOXA10调控DNM1L表达机制的研究[J].南京农业大学学报,2016, 39(6):1023-1029.XIAO C,HU J,JI H J,et al.The regulation mechanism of HOXA10 on DNM1L expression in mouse endometrial stromal cells[J].Journal of Nanjing Agricultural University,2016,39(6):1023-1029.(in Chinese)
[34]	HOPPINS S,LACKNER L,NUNNARI J.The machines that divide and fuse mitochondria[J].Annu Rev Biochem,2007, 76:751-780.
[35]	MAMMOTO A,MUYLEART M,KADLEC A,et al.YAP1-TEAD1 signaling controls angiogenesis and mitochondrial biogenesis through PGC1α[J].Microvasc Res,2018,119:73-83.
[36]	安世钰,张国敏,尤佩华,等.PPARGC1A/NRF1/TFAM通路核心基因在湖羊睾丸发育过程中的表达变化[J].南京农业大学学报,2019,42(2):345-351.AN S Y,ZHANG G M,YOU P H,et al.Research on the change of PPARGC1A/NRF1/TFAM pathway genes expression in the development of Hu sheep testis[J].Journal of Nanjing Agricultural University,2019,42(2):345-351.(in Chinese).

Expression Patterns and Functional Analysis of YAP1 in Endometrium of Hu Sheep with Different Fertility during Estrus

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 36

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Linli, PENG Xuelan, LI Bo, CHENG Lefan, CIREN Lamu, ZHANG Enping. Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3275-3285.
[2]	XIONG Chengkun, ZHANG Daoliang, YANG Yue, DING Hongyan, ZHAO Jie, LI Yu, WANG Xichun, FENG Shibin, ZHAO Chang, TANG Jishun, WU Jinjie. Effect of Rutin on Rumen Fermentation, Rumen Flora Structure and Antioxidant Properties in Perinatal Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2898-2909.
[3]	YE Qianwen, CHEN Zhuo, LI Xin, SUN Yawei, JIN Xiaoye, LI Ziqian, WUMAIERJIANG·Yahefu, ZHONG Qi, MA Xuelian, YAO Gang. Dynamic Changes of Gut Microbiota Composition and Its Predicted Metabolism Function in One-month-old Sucking Lambs [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1095-1108.
[4]	WEI Xiao, ZHANG Jiantong, LONG Tanghui, LI Kairong, LI Yanjiao, OUYANG Kehui, QIU Qinghua. Effects of Dietary Energy Level on Rumen Fermentation Characteristics and Microbial Composition of Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3042-3051.
[5]	LI Yu, DUAN Chunhui, SONG Zhipan, YUE Sicong, WANG Yuan, ZHANG Yingjie, LIU Yueqin. Effects of PGF2α on Reproductive Hormones and Related Cytokines during Luteal Phase in Ewes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1807-1818.
[6]	YANG Hua, ZHAO Xinyue, WAN Zhen, WANG Feng, ZHANG Yanli. Effect of HSD17B12 Gene on Gonadotropin Synthesis in Hu Sheep Pituitary Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 703-713.
[7]	GUO Xiaoxiao, LI Yinxia, WANG Yue, ZHANG Han, ZHANG Jun, QIAN Yong, MENG Chunhua, WANG Huili, ZHONG Jifeng, CAO Shaoxian. Polymorphisms in the 5'Regulatory Region of PLAG1 Gene and Their Association with Early Body Weight of Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 331-343.
[8]	Lü Songjie, FU Li, FAN Wenhua, ZHANG Shushan, WU Caifeng, XU Jiehuan, DAI Jianjun, ZHANG Defu. Protective Effect of Mitochondria-targeted Antioxidant Mitoquinone on the Damage of Hu Sheep Frozen Sperm [J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50(12): 2554-2559.
[9]	ZHAO Guohong, WANG Shiqin, WANG Fen, WANG Hong, DIAO Qiyu, HAN Xiangmin, ZHANG Naifeng. Effects of Yeast Culture Supplementation in High-Concentration Diet on Nutrient Digestibility and Rumen Fermentation of Fattening Hu Sheep [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2156-2165.
[10]	FAN Yi-xuan, DENG Kai-ping, TANTAI Wen-jing, HUANG Xin-ai, WANG Feng, CHEN Ming. Effect of Selenium Yeast in Containing PUFAs Dietary on Fatty Acid Composition and Antioxidant Capacity of Hu Lambs [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1661-1673.
[11]	YAO Xiao-lei, HUANG Xin-ai, XIAO Shen-hua, ZHENG Lin-feng, FAN Li-jie, JIN Yu-yue, LIU Zi-fei, ZHANG Yan-li, WANG Jie, WANG Feng. Expression Patterns and Correlation of Adiponectin Receptors and Testosterone Secretion Related-genes Expression in Male Reproductive Organs at Different Developmental Stages of Hu Sheep [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1642-1650.
[12]	WANG De-di, LI Yin-xia, YAO Yi-long, PAN Zeng-xiang, JUE Ken, YI Ming, CAO Shao-xian, LI Qi-fa. Transcription Factor PAX4 Regulates the Transcriptional Activity of FSHR Gene in Hu Sheep [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 919-926.
[13]	NIE Hai-tao, HUANG Xin-ai, LIU Hao, GUO Yi-xuan, WANG Zi-yu, ZHANG Yan-li, WAN Yong-jie, FAN Yi-xuan, ZHANG Guo-min, WANG Feng, WANG Jie. Effect of Different Nutrient Levels on Follicle Development and Glucose Metabolic Pathway Involved in Folliculogenesis of Hu Sheep during Luteal Phase [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 942-953.
[14]	LI Xue-ling, ZHANG Nai-feng, MA Tao, TAO Da-yong, CHAI Jian-min, WANG Yu-rong, ZHANG Fan, DIAO Qi-yu. Effects of Lysine, Methionine, Threonine and Tryptophan in Starter on Growth Performance, Nitrogen Utilization and Serum Parameters in Weaned Lambs [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(4): 678-689.
[15]	ZHANG Fan,CUI Kai, WANG Jie,DIAO Qi-yu. Effects of Dietary Nutrient Levels on the Development of Late Pregnant Ewes and Fetal [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 474-482.