卵母细胞成熟相关激素和生长因子受体在马扩展型和紧凑型卵丘-卵母细胞复合体表达的研究  

Differential Expression of Oocyte Development-related Hormone and Growth Factor Receptors in Equine Expanded and Compact Cumulus-oocyte Complexes

作　　者：神英超 陶力 任宏 王希生 田书岳 杜明 芒来[1] 格日乐其木格 SHEN Yingchao;DAVSHILT Toli;REN Hong;WANG Xisheng;TIAN Shuyue;DU Ming;DUGARJAVIIN Manglai;BOU Gerelchimeg(Inner Mongolia Key Laboratory of Equine Genetics,Breeding and Reproduction,College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学动物科学学院、内蒙古自治区马遗传育种与繁殖重点实验室,呼和浩特010018

出　　处：《畜牧兽医学报》2023年第9期3735-3744,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：内蒙古自治区直属高校基本科研业务费项目(BR220402);中央引导地方科技发展资金项目(2022ZY0132);内蒙古自治区自然科学基金项目(2020BS03035)。

摘　　要：旨在比较一些与卵细胞成熟相关的激素和生长因子受体在两种卵丘-卵母细胞复合体(Ex-COC和Cp-COC)中的表达以探究两种类型的马卵母细胞成熟和发育能力差异的原因。本研究在屠宰场采集蒙古马离体卵巢带回实验室回收COCs,并根据卵丘细胞形态区分Ex-和Cp-COCs;通过qPCR和免疫荧光检测FSHR、LHR、IGF1R、IGF2R、ESR1、ESR2,BMP15和GDF9受体BMPR1B、BMPR2和ALK5在两种COCs中的表达模式;将雌二醇、IGF2、GDF9和BMP15分别添加进马IVM培养体系中检测其对马卵母细胞体外成熟率的影响。结果显示,这些受体基因的表达量在Ex-和Cp-COCs的卵丘细胞和卵母细胞中均有差异,在Cp-COCs中,卵丘细胞的大部分受体基因表达量高于卵母细胞;而在Ex-COCs中,卵母细胞与卵丘细胞的基因表达水平相似或高于卵丘细胞。相较于Cp-COCs的卵母细胞,大部分的受体基因在Ex-COCs的卵母细胞中表现出更高表达水平。体外成熟培养试验表明雌二醇和IGF2的添加可能有利于提高马的卵母细胞体外成熟率。马扩展型和紧凑型COCs中FSH、LH、IGF1、IGF2、雌激素、BMP15和GDF9等重要激素或生长因子受体表达水平的差异可以解释其体外成熟和发育能力的差异,在成熟培养体系中添加IGF2和E2可能会提高马两种类型卵母细胞体外成熟率。The present study was conducted to compare the expression of some growth factors and hormone receptors associated with oocyte maturation in Ex-COC and Cp-COC to investigate the reasons for the difference in maturation and development capacity in the two types of equine oocytes.Isolated ovaries from Mongolian horses were collected at the slaughterhouse and brought back to the laboratory to recover COCs,Ex-and Cp-COCs were distinguished according to cumulus morphology;the expression pattern of FSHR,LHR,IGF 1R,IGF 2R,ESR1,ESR 2,the BMP15 and GDF9 receptors BMPR 1B,BMPR 2,and ALK 5 were studied by qPCR and immunofluorescence in both COCs.Estradiol,IGF2,GDF9,and BMP15 were added to the equine IVM culture system to examine their effects on the maturation rate of equine oocytes in vitro.The results showed that the expression of these receptor genes was difference in cumulus cells and oocytes between Ex-and Cp-COCs,with most receptor genes being expressed at higher levels in cumulus than in oocytes in Cp-COCs,while in Ex-COCs,the levels in cumulus were similar with oocytes or higher in oocytes.Most of the receptor genes showed higher expression levels in oocytes from Ex-COCs compared to oocytes from Cp-COCs.and the differential expression of growth factor and hormone receptor genes might be responsible for the difference in developmental capacity.In vitro maturation culture experiments revel that the addition of estrogen and IGF2 may be beneficial in increasing the rate of maturation of equine oocytes in vitro.Different expression levels of the important hormones or growth factor receptors such as FSH,LH,IGF1,IGF2,estrogen,BMP15,and GDF9 in equine Ex-and Cp-COCs could explain the differences in maturation and developmental capacity in vitro,and the addition of IGF2 and E2 to the maturation culture system may increase the in vitro maturation rate of equine oocytes.

关 键 词：马卵母细胞 促卵泡素(FSH) 胰岛素生长因子1/2(IGF1/2) 雌激素 生长分化因子9(GDF9) 骨形态发生蛋白15(BMP15) 体外成熟 

分 类 号：S821.2[农业科学—畜牧学]