PLCγ1基因对绵羊早期胚胎体外发育的影响

doi:10.11843/j.issn.0366-6964.2022.08.016

Abstract

Abstract: The purpose of this study was to investigate the effect of PLCγ1 on the development of sheep early embryo in vitro, and to lay a foundation for revealing the action mechanism of PLCγ1 in the development of sheep early embryo. In this study, sheep oocytes with more than 3 layers of granulosa cells were selected as experimental materials, and they were divided into two groups after mature in vitro culture. PLCγ1 was introduced into MⅡ oocytes (n=127) by microinjection as experimental group. Microinjection of empty vector pcDNA3.1-EGFP was used as the control group (n=120), and after 48 h the cleavage rate were measured. The Ca²⁺ probe Rhod 2-AM was used to monitor the Ca²⁺ fluctuation of MⅡ oocytes and early embryos at 16, 48, 72, 96 and 120 h after microinjection. The results showed that the cleavage rate was ((19.67±0.2)%, P<0.01)) and morula development rate was ((9.00±0.17)%,P<0.05)) after microinjection of PLCγ1. After injection of PLCγ1, Ca²⁺ concentration in oocytes and early embryos varied at different developmental stages.It was observed that Ca²⁺ was mainly distributed in the cytoplasm and reached the peak at 48 h after injection (P<0.01). The results showed that PLCγ1 gene could induce Ca²⁺ oscillation during ovine oocyte development, initiate cleavage, and promote the development of early embryo.

Key words: PLCγ1, Ca²⁺, oocyte, morula, sheep

CLC Number:

S826.3

WU Xiaoxue, YUAN Liming, LIU Xinjie, LIU Suping, CHEN Ning, SAIWU Jiafu. Effect of PLCγ1 Gene on the Development of Sheep Early Embryo in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2578-2585.

References 35

[1]	周荣, 杨秀萍.卵母细胞减数分裂的调控机制研究进展[J].国际生殖健康/计划生育杂志, 2008, 27(2):80-84.ZHOU R, YANG X P.Recent advances in understanding the regulation of oocyte meiosis[J].Journal of International Reproductive Health/Family Planning, 2008, 27(2):80-84.(in Chinese)
[2]	ZHANG M J, SU Y Q, SUGIURA K, et al.Granulosa cell ligand NPPC and its receptor NPR2 maintain meiotic arrest in mouse oocytes[J].Science, 2010, 330(6002):366-369.
[3]	SUN L, HODEIFY R, HAUN S, et al.Ca2+homeostasis regulates Xenopus oocyte maturation[J].Biol Reprod, 2008, 78(4):726-735.
[4]	刘红林, 汪河海, 范必勤, 等.显微注射Ca2+激活小鼠MⅡ期卵母细胞[J].畜牧兽医学报, 1997, 28(3):206-210.LIU H L, WANG H H, FAN B Q, et al.Activation of mouse oocytes (M Ⅱ) by intracellular microinjection of calcium[J]. Acta Veterinaria et Zootechnica Sinica, 1997, 28(3):206-210.(in Chinese)
[5]	JANG H J, SUH P G, LEE Y J, et al.PLCγ1:potential arbitrator of cancer progression[J].Adv Biol Regul, 2017, 67:179-189.
[6]	WOO S J, JO H I, LEE H H, et al.Molecular characterization and expression analysis of olive flounder (Paralichthys olivaceus) phospholipase C gamma 1 and gamma 2[J].Fish Shellfish Immunol, 2017, 63:353-366.
[7]	JANG H J, SUH P G, LEE Y J, et al.PLCγ1:potential arbitrator of cancer progression[J].AdvBiol Regul, 2018, 67:179-189.
[8]	MANDAL S, BANDYOPADHYAY S, TYAGI K, et al.Recent advances in understanding the molecular role of phosphoinositide-specific phospholipase C gamma 1 as an emerging onco-driver and novel therapeutic target in human carcinogenesis[J].Biochim Biophys Acta Rev Cancer, 2021, 1876(2):188619.
[9]	MALCUIT C, KNOTT J G, HE C L, et al.Fertilization and inositol 1, 4, 5-trisphosphate (IP3)-induced calcium release in type-1 inositol 1, 4, 5-trisphosphate receptor down-regulated bovine eggs[J].Biol Reprod, 2005, 73(1):2-13.
[10]	郝肖琼, 贾方毅, 付旭阳.PLCγ1在小鼠卵母细胞减数分裂恢复中的作用[J].中国比较医学杂志, 2019, 29(12):16-22, 51.HAO X Q, JIA F Y, FU X Y.The role of PLCγ1 in mouse oocyte meiotic resumption[J].Chinese Journal of Comparative Medicine, 2019, 29(12):16-22, 51.(in Chinese)
[11]	RUNFT L L, CARROLL D J, GILLETT J, et al.Identification of a starfish egg PLC-γ that regulates Ca2+ release at fertilization[J].Dev Biol, 2004, 269(1):220-236.
[12]	FINLEY J.Oocyte activation and latent HIV-1 reactivation:AMPK as a common mechanism of action linking the beginnings of life and the potential eradication of HIV-1[J].Med Hypotheses, 2016, 93:34-47.
[13]	FERRER-BUITRAGO M, BONTE D, DESUTTER P, et al.Single Ca2+transients vs oscillatory Ca2+signaling for assisted oocyte activation:limitations and benefits[J].Reproduction, 2018, 155(2):R105-R119.
[14]	KRAWCZYK P, MATUSZYK J.The mechanism of phospholipase Cγ1 activation[J].Postepy Hig Med Dosw(Online), 2011, 65:470-477.
[15]	FEITOSA W B, LOPES E, VISINTIN J A, et al.Endoplasmic reticulum distribution during bovine oocyte activation is regulated by protein kinase C via actin filaments[J].J Cell Physiol, 2020, 235(7-8):5823-5834.
[16]	KAUFMAN M H.Parthenogenetic activation of oocytes[J/OL].Cold Spring Harb Protoc, 2018, 2018(1), doi:10.1101/pdb.prot094409.
[17]	SINGH K P, MOHAPATRA S K, KAUSHIK R, et al.Parthenogenetic activation of buffalo (Bubalusbubalis) oocytes:comparison of different activation reagents and different media on their developmental competence and quantitative expression of developmentally regulated genes[J].Zygote, 2021, 29(1):49-58.
[18]	LAN G C, HAN D, WU Y G, et al.Effects of duration, concentration, and timing of ionomycin and 6-dimethylaminopurine (6-DMAP) treatment on activation of goat oocytes[J].Mol Reprod Dev, 2005, 71(3):380-388.
[19]	MEHLMANN L M, CARPENTER G, RHEE S G, et al.SH2 Domain-mediated activation of phospholipase Cγis not required to initiate Ca2+release at fertilization of mouse eggs[J].Dev Biol, 1998, 203(1):221-232.
[20]	袁瑞莹.不同处理方式对小鼠卵母细胞孤雌激活钙离子振荡的影响[D].广州:南方医科大学, 2018.YUAN R Y.Effects of different treatments on the[Ca2+] i oscillations inparthenogenetic activation of mouse oocyte[D].Guangzhou:Southern Medical University, 2018.(in Chinese)
[21]	YESTE M, JONES C, AMDANI S N, et al.Oocyte activation and fertilisation:crucial contributors from the sperm and oocyte[J].Results Probl Cell Differ, 2017, 59:213-239.
[22]	SILVESTRE F, BONI R, FISSORE R A, et al.Ca2+signaling during maturation of cumulus-oocyte complex in mammals[J].Mol Reprod Dev, 2011, 78(10-11):744-756.
[23]	TAROZZI N, NADALINI M, COTICCHIO G, et al.The paternal toolbox for embryo development and health[J].Mol Hum Reprod, 2021, 27(7):gaab042.
[24]	FINKELSTEIN M, ETKOVITZ N, BREITBART H.Ca2+signaling in mammalian spermatozoa[J].Mol Cell Endocrinol, 2020, 516:110953.
[25]	MARTIN J H, BROMFIELD E G, AITKEN R J, et al.Biochemical alterations in the oocyte in support of early embryonic development[J].Cell Mol Life Sci, 2017, 74(3):469-485.
[26]	SETTE C, BEVILACQUA A, GEREMIA R.Involvement ofphospholipase Cγ1 in mouse egg activation induced by a truncated form of the C-kit tyrosine kinase present in spermatozoa[J].J Cell Biol, 1998, 142(4):1063-1074.
[27]	PAN B, LI J L.The art of oocyte meiotic arrest regulation[J].Reprod Biol Endocrinol, 2019, 17(1):8.
[28]	ANIFANDIS G, MICHOPOULOS A, DAPONTE A, et al.Artificial oocyte activation:physiological, pathophysiological and ethical aspects[J].Syst Biol Reprod Med, 2019, 65(1):3-11.
[29]	PARRINGTON J, ARNOULT C, FISSORE R A.Theeggstraordinary story of how life begins[J].Mol Reprod Dev, 2019, 86(1):4-19.
[30]	YANG F L, HUANG L, TSO A, et al.Inositol 1, 4, 5-trisphosphate receptors are essential for fetal-maternal connection and embryo viability[J].PLoS Genet, 2020, 16(4):e1008739.
[31]	LIANG S H, ZHAO J, WANG Q Y, et al.Carbon monoxide enhances calcium transients and glucose-stimulated insulin secretion from pancreatic β-cells by activating phospholipase C signal pathway in diabetic mice[J]. Biochem Biophys Res Commun, 2021, 582:1-7.
[32]	SETTE C, PARONETTO M P, BARCHI M, et al.Tr-kit -induced resumption of the cell cycle in mouse eggs requires activation of a Src-like kinase[J].EMBO J, 2002, 21(20):5386-5395.
[33]	杨霖, 罗深秋.PLC-γ1对PDGF介导的细胞内钙流的影响[J].中国医学物理学杂志, 2001, 18(2):93-94, 100.YANG L, LUO S Q.The effect of PLC-γ1on mobilization of intracellular Ca2+ induced by PDGF[J].Chinese Journal of Medical Physics, 2001, 18(2):93-94, 100.(in Chinese)
[34]	YIN X Q, ECKBERG W R.Characterization of phospholipases C β and γ and their possible roles in Chaetopterus egg activation[J].Mol Reprod Dev, 2010, 76(5):460-470.
[35]	WU H, SMYTH J, LUZZI V, et al.Spermfactor induces intracellular free calcium oscillations by stimulating the phosphoinositide pathway[J].Biol Reprod, 2001, 64(5):1338-1349.

Effect of PLCγ1 Gene on the Development of Sheep Early Embryo in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 35

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LI Wanjun, XU Jiehuan, HE Mengxian, KONG Yuting, ZHANG Defu, DAI Jianjun. Cytochalasin B Alleviates the Migration Disorder of Cortical Particle Caused by Vitrification in Porcine Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1999-2010.
[2]	PENG Peiya, CHEN Yuhan, YANG Long, WANG Ming, ZHAO Ruiting, HE Jun, YIN Yulong, LIU Mei. Research Progress of Copy Number Variation in Livestock [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1356-1369.
[3]	ZHANG Shaohua, WANG Shuai, ZOU Yang, LIU Zhongli, CAI Xuepeng. Advances in Detection Approaches for Ovine Haemonchosis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1499-1510.
[4]	YANG Yang, YU Qian, LIU Yucheng, YANG Hua, ZHAO Zhuo, WANG Limin, ZHOU Ping, YANG Qingyong, DAI Rong. Identification and Tissue Expression Analysis of the Sheep MYL Gene Family [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1551-1564.
[5]	LAN Xinrui, ZHAO Baobao, ZHANG Bihan, LIN Xiaoyu, MA Huiming, WANG Yongsheng. Effects of β-sitosterol on Porcine Oocyte Maturation and Embryonic Development in Vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1629-1637.
[6]	LI Yujun, HE Honghong, YANG Lixue, YANG Xiaogeng, LI Jian, ZHANG Huizhu. Advances in Regulation of Mammalian Embryonic Development by Mitochondrial Autophagy [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 905-912.
[7]	CHANG Xindan, HU Fan, WU Zhiwu, YE Bingsen, LIU Tiehai, LIN Jie, HE Zhixiong, TAN Zhiliang. Effect of High Proportion Rumen Bypass Fat Diet on Feeding Behavior of Growing Mutton Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1077-1084.
[8]	GAO Hui, FANG Min, JIANG Lingling, MA Yaoyu, LIU Qiang, ZHANG Gang, NIU Xiaoxia, WANG Pu, LI Yong, ZHANG Sinong. Meta-analysis of Bluetongue Virus Prevalence in Sheep Flocks in China from 2012—2022 [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 706-717.
[9]	WANG Haibo, ZHAN Jinshun, GU Zhiyong, CHEN Xinfeng, PAN Yue, JIA Haobin, ZHONG Xiaojun, LI Kairong, ZHAO Shengguo, HUO Junhong. Comparative Study on Meat Quality Characteristics of Three-Way Hybrid Sheep Charolais×Duper×Hu and Charolais×Australian White×Hu and Hu Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 110-119.
[10]	DUAN Xiangru, KANG Jia, YANG Ruochen, SHAN Xinyu, LI Taichun, ZHAO Wen, ZHANG Yingjie, LIU Yueqin. Effect of L-cysteine on Proliferation, Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 179-191.
[11]	SHEN Yingchao, DAVSHILT Toli, REN Hong, WANG Xisheng, TIAN Shuyue, DU Ming, DUGARJAVIIN Manglai, BOU Gerelchimeg. Differential Expression of Oocyte Development-related Hormone and Growth Factor Receptors in Equine Expanded and Compact Cumulus-oocyte Complexes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3735-3744.
[12]	ZHANG Hanyue, ZHAO Dan, LIANG Yu, ZHAO Bishi, FAN Mengdan, QIAO Liying, LIU Jianhua, YANG Kaijie, PAN Yangyang, LIU Wenzhong. miR-150 Regulates Ovine Preadipocyte Differentiation by Targeting AOC3 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3262-3274.
[13]	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.
[14]	HE Mingyang, MA Yujing, WANG Yong, YANG Ruochen, LIU Yueqin, ZHANG Yingjie, DUAN Chunhui. Effects of Melatonin on Proliferation, Apoptosis of Ovarian Granulosa Cells, and Its Secretion of Steroid Hormones of Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3313-3324.
[15]	SONG Meijun, HAO Kexing, HAI Siyu, CHEN Yan, WANG Jing, HU Guangdong. Effects of SRIF-14 on Proliferation and Apoptosis of Endometrial Epithelial Cells in Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3325-3334.