过表达FOXL2基因对鸡胚性腺分化的影响

doi:10.16431/j.cnki.1671-7236.2020.09.019

摘要/Abstract

摘要： 试验旨在研究FOXL2基因对鸡胚性腺分化的影响。本试验分为试验1组、试验2组和空白对照组（鸡胚数量分别为260、100、20枚），试验组通过胚盘下腔注射的方法分别将pLV-FOXL2慢病毒重组质粒、pLV空质粒注入胚胎期第2天的鸡胚，空白对照组不做处理并与试验组一起孵化至出雏，利用CHD1基因遗传性别鉴定的方法对出雏的雏鸡进行性别检测，分析其性腺解剖学、组织学结构变化，并利用免疫组化的方法检测性腺FOXL2和CYP19A1蛋白表达量。结果显示，试验1组遗传性别为公的23只，遗传性别为母的18只，表型性别为公的21只，表型性别为母的18只，其中有2只表型性别不典型，左侧性腺发生变化，朝卵巢结构转变；试验2组遗传性别为公的9只，遗传性别为母的12只，表型性别与遗传性别一致。阳性PCR检测结果显示，试验1组获得阳性个体10个，阳性率为24.4%（10/41）；试验2组获得阳性个体8个，阳性率为38.1%（8/21）。性腺解剖学结果显示，阳性pLV-FOXL2雄性鸡胚左侧性腺体积明显大于右侧性腺，表现膨松状态；组织切片结果显示，雄性鸡胚性腺具有典型的卵巢皮质层和髓质层结构；阳性pLV-FOXL2雌性鸡胚性腺的发育无明显变化。免疫组化结果显示，FOXL2和CYP19A1蛋白在试验1组左右侧睾丸中的表达量与空白对照组母鸡卵巢中的表达量相似，显著高于试验2组（P<0.05）。以上结果表明，FOXL2基因可能促进鸡雄性性腺的性反转，在鸡性腺分化和发育过程中发挥着重要的作用。

关键词: 鸡胚; 性腺分化; FOXL2基因; 性别相关基因

Abstract: The aim of this study was to investigate the effect of FOXL2 gene on gonadal differentiation in chicken embryos.This test was divided into the experiment groups 1,2 and blank control group(chicken embryos were 260,100 and 20,respectively).In the experimental group,pLV-FOXL2 lentivirus recombinant plasmids and pLV empty plasmids were injected into the chicken embryo on the second day of the embryonic stage by subpaneoidal injection.The blank control group was not treated and incubated with the experimental groups until the chick was hatched.CHD1 gene genetic sex identification method was used to detect the sex of chicks,the changes of gonadal anatomy and histological structure were analyzed,and the expression levels of FOXL2 and CYP19A1 proteins were detected by immunohistochemistry.The results showed that in the experiment group 1,there were 23 male and 18 female of genetic sex,21 male and 18 female of phenotypic sex,and two of them had atypical phenotypic gender,with changes in the left gonadal gland toward ovarian structure.The phenotypic sex was consistent with the genetic sex in experiment group 2,with 9 male and 12 female.Positive PCR results showed that 10 positive individuals were obtained in the experiment group 1,with a positive rate of 24.4% (10/41),and 8 positive individuals were obtained in the experiment group 2,with a positive rate of 38.1% (8/21).The anatomical structure of the gonad showed that the volume of the left gonad was significantly larger than that of the right gonad in the positive plV-FOXL2 male embryos.The results of tissue sections showed that the gonad of male chicken embryo had typical structure of ovarian cortex and medulla.There was no significant change in the development of gonad in female embryos with positive pLV-FOXL2.Immunohistochemical results showed that the expression levels of FOXL2 and CYP19A1 proteins in the left and right testicle of experiment group 1 were similar to that in the hen ovary of the blank control group,and were significantly higher than that in experiment group 2 (P<0.05).These results suggested that FOXL2 gene might promote the sexual inversion of chicken gonads and played an important role in the differentiation and development of chicken gonads.

Key words: chicken embryo; gonadal differentiation; FOXL2 gene; sex related genes

中图分类号:

S831.2

徐文文, 夏利, 孙甜甜, 邓继贤, 杨祝良, 杨秀荣. 过表达FOXL2基因对鸡胚性腺分化的影响[J]. 中国畜牧兽医, 2020, 47(9): 2857-2865.

XU Wenwen, XIA Li, SUN Tiantian, DENG Jixian, YANG Zhuliang, YANG Xiurong. Effect of Overexpression of FOXL2 Gene on Gonadal Differentiation in Chicken Embryo[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2857-2865.

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