XY性反转雌性小鼠的鉴定和初步分析

doi:10.11843/j.issn.0366-6964.2019.11.008

摘要/Abstract

摘要： 旨在繁育并鉴定XY雌性小鼠，初步分析其生殖能力和性反转的机制，为小鼠发育过程中性别决定相关研究提供动物模型。XY雌性小鼠的鉴定分为表型鉴定和基因型鉴定两步：令B6.XY^TIR雄性小鼠与野生型B6雌性小鼠自然交配，选用30、60、90日龄仔鼠，观察其外生殖器特征和肛殖距以鉴定雌雄；处死交配后17.5天的孕鼠，取胎鼠，根据其性腺形态学和生殖细胞类型鉴定雌雄（表型鉴定）。提取仔鼠或胎鼠DNA，设计引物扩增Zfy基因（Y染色体上的特有基因），以确定其性染色体为XX或XY（基因型鉴定），结合表型鉴定，从而得到B6.XY^TIR雌性小鼠。本研究同时用全组织免疫荧光染色的方法分析胎儿期性腺的分化，用形态学观察的方法观察出生后小鼠内生殖系统，通过测序对Zfy基因的序列进行初步分析。本研究通过表型鉴定和基因型鉴定得到了一种XY性反转雌性小鼠；通过对出生小鼠内生殖系统的观察，发现其拥有与野生型XX雌鼠相似的卵巢、输卵管和子宫结构；同时初步研究了性腺分化过程中生殖细胞的分布，发现XY雌性胎鼠生殖细胞分布类型与野生型类似；在扩增Zfy基因进行基因型鉴定的同时，测序表明B6.XY^TIR的后代中，XY雄性和XY雌性均只有Zfy-1的基因，而未能扩增到Zfy-2基因；XY雌鼠在所有XY小鼠（99只）中所占的比例为47.47%。XY雌性小鼠拥有一套完整的雌性生殖系统，且性反转在胚胎发育过程中已经确立；B6.XY^TIR的Y染色体上可能缺失Zfy-2基因，推测其与性反转的发生有关系；本试验结果可为深入研究性腺分化和生殖细胞发生提供独特的视角。

Abstract: This study aimed to breed and identify XY female mice, and analyze their reproductive ability and the mechanism of sexual inversion, so as to provide an animal model for studying on gender determination during mouse development. The identification method of XY female mice was divided into two steps(phenotypic and genotypic identification). The first, phenotypic identification:B6.XY^TIR male was naturally mated with wild-type XX female mice, their offspring were analyzed. Male and female mice of 30-, 60- and 90-day-old were identified by external genital characteristics and anogenital distance, and the gender of fetuses of 17.5 days post coitum were identified by gonadal morphology and germ cell types. The second, genotype identification:In order to determine their sex chromosome as XX or XY, DNA of postnatal and fetal mice was extracted, primers were designed and Zfy gene (a unique gene on the Y chromosome) was amplified. Combined with phenotypic identification, B6.XY^TIR female mice were obtained. In this study, whole-tissue immunofluorescence staining was used to analyze the differentiation of gonads in fetal stage, and morphological observation was used to observe the internal reproductive system of postnatal mice, and the sequence of Zfy gene was preliminarily analyzed by sequencing. A type of XY sex reversed female mouse was obtained through phenotypic and genotypic identification. It was found by observation that the ovaries, fallopian tubes and uterine structures of the XY female mice were similar to those of wild-type XX female mice. Meanwhile, the preliminary study showed that the distribution of germ cells during gonad differentiation in XY female mice was similar to that in the wild-type XX female. Sequencing showed that only Zfy-1 gene, but not Zfy-2 gene, was amplified in B6.XY^TIR males and B6.XY^TIR females; XY females accounted for 47.47% of all 99 XY mice. The results showed that XY females had a complete female reproductive system, and sex reversed had already been formed during embryonic development. Zfy-2 might be missing on the Y chromosome of B6.XY^TIR female mice, which could be related to the occurrence of sexual inversion. The results provides a unique perspective for the further study of gonadal differentiation and gametogenesis.

中图分类号:

S814

曹江琴, 刘艳芳, 黄竹涛, 陈佳莉, 史悦, 贺思羽, 刘宗平, 朱家桥. XY性反转雌性小鼠的鉴定和初步分析[J]. 畜牧兽医学报, 2019, 50(11): 2244-2251.

CAO Jiangqin, LIU Yanfang, HUANG Zhutao, CHEN Jiali, SHI Yue, HE Siyu, LIU Zongping, ZHU Jiaqiao. Identification and Preliminary Analysis of XY Sex Reversed Female Mice[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(11): 2244-2251.

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