Tle4z1 Facilitate the Male Sexual Differentiation of Chicken Embryos.
Tle4z1 Facilitate the Male Sexual Differentiation of Chicken Embryos.
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Tle4z1 促进鸡胚胎的雄性分化
DOI：
10.3389/fphys.2022.856980
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发表时间：
2022
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
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鸡的性别分化是一个复杂的、受多基因精确调控的过程。然而，关于性别分化的关键基因仍不清楚。目的：探讨RNAseq测序筛选到的Tle4z1基因在鸡胚胎发育过程中性别分化的作用。Tle4z1在公鸡和母鸡的ESCs和PGCs的RNA-seq中有差异表达。然后，我们建立了一种有效的方法，分别在体内和体外过表达或下调Tle4z1的表达。应用组织形态观察、qRT-PCR和ELISA法检测Tle4z1基因在第0天胚胎注射载体的雄性分化过程中的作用。结果表明，Tle4z1在胚胎4.5天有明显的雄性偏好，这种现象持续存在于鸡胚胎的整个生长期。形态观察结果表明，Tle4z1基因敲除的遗传雄性(ZZ)胚胎两侧性腺发育不对称，性腺皮质增厚，具有典型的遗传雌性(ZW)性腺特征。此外，控制雌性分化的CYP19A1的表达显著增加，而雄性标记基因DMRT1、Sox9、WT1和AR的表达显著下调。此外，睾酮水平也显著降低，与Tle4z1的表达呈正相关(P<；0.01)。相反，当Tle4z1过表达时，ZW胚胎表现出去雌雄发育。我们通过基因调控过程和睾酮的合成，证明Tle4z1是一个雄性分化的新基因，为理解鸡性别分化的分子机制奠定了基础。
Sex differentiation is a complex and precisely regulated process by multiple genes in chicken. However, it is still unclear on the key genes of sex differentiation. To explore the function of Tle4z1 screened by RNA-seq sequencing on sex differentiation during the development of chicken embryos. Tle4z1 was differentially expressed from the RNA-seq of ESCs and PGCs in male and female chickens. Then, we established an effective method to overexpression or knocking down the expression of Tle4z1 in ovo and in vitro, respectively. Histomorphological observation, qRT-PCR and ELISA were applied to detect the function of Tle4z1 in the process of male sex differentiation by injecting vectors into embryos at day 0. It showed that Tle4z1 has significant male preference in embryonic day 4.5, such phenomenon persisted during the growth period of chicken embryos. Morphological observation results showed that the gonads on both sides of genetic male (ZZ) embryos with Tle4z1 knocking down developed asymmetrically, the gonadal cortex became thicker showing the typical characteristics of genetic female (ZW) gonads. Furthermore, the expression of Cyp19a1, which dominates female differentiation, was significantly increased, while the expression of male marker genes Dmrt1, Sox9, WT1 and AR was significantly downregulated. In addition, the concentration of testosterone also significantly decreased, which was positively correlated with the expression of Tle4z1 (P < 0.01). Conversely, the ZW embryo showed defeminized development when Tle4z1 was overexpressed. We prove that the Tle4z1 is a novel gene through the male sexual differentiation via gene regulation process and synthesis of testosterone, which construct the basis for understanding the molecular mechanism of sex differentiation in chickens.
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DOI：
10.3390/ani10122228
发表时间：
2020-11-27
期刊：
Animals : an open access journal from MDPI
影响因子：
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