哺乳动物性别决定机制的研究进展

哺乳动物的性别决定包括初级性别决定和次级性别决定,是以SRY基因为主导,其他多个基因参与的级联调控过程。近年的研究表明。SRY、DAX1、SOX3等性染色体基因和SOX9、MIS、WT1、SF1等常染色体基因都参与性别决定的级联过程。结合中学生物学教材及发育生物学相关原理,从性染色体上和常染色体上与性别决定有关的基因阐述哺乳动物的性别决定机制,并简述了哺乳动物的性别决定模型。     

WT1在哺乳动物性别发育过程中的调控作用

哺乳动物的性别发育大致可分为性别决定和性别分化两步,是一个由WT1/Wt1、SRY/Sry和MIS/Mis等多基因参与的级联过程,但目前对于这些基因之间的相互作用尚不清楚。在性别发育过程中持续表达的WT1/Wt1与多种伴有性别发育异常的疾病相关,其重要性表现为对多个性别发育关键基因在转录水平和转录后水平的调控。简要概述了WT1/Wt1的复杂性及其对多基因的调控作用,以期为阐明性别发育机制和基因间的相互关系提供参考。     

江豚Sry基因和Dmrt家族六个基因的序列分析

Sry基因是哺乳动物性别决定的开关基因,其功能是调控下游Dmrt、Sox3、Sox9等基因来启动性别的分化过程。Dmrt家族是一个在性别决定和分化发育中具有重要功能的基因家族,该家族成员都含有一个具有DNA结合能力的保守基序-DM结构域。本文通过PCR及克隆获得了江豚的Sry基因全序列;通过PCR-SSCP法筛选获得了江豚的Dmrt家族六个不同DM序列。结果显示,雄性个体中存在Sry基因,而Dmrt基因在雌雄个体中都有,与其他动物相关基因进行聚类分析,显示它们在进化上具有高度的保守性。     

哺乳动物性别决定及其机制的研究

哺乳动物的性别决定与性别分化是在以SRY基因为主,SOX基因、DAX-1等众多基因的参与下实现的。哺乳动物性别决定的研究涉及到三个方面的内容:性别决定机制、性别决定进化和物种间性别决定差异。性别决定的进化和物种间性别决定的差异对性别决定机制的研究提供了重要的线索。     

昆虫性别决定机制研究进展

阐述昆虫的性别决定机制是理解昆虫性别分化调控的理论基础,也为人类有效控制害虫开辟了新方向。昆虫性别决定机制存在复杂性和多样性,但主要是内因即性别决定基因级联互作调控的结果。本文对近年来基于性别决定基因级联互作的昆虫性别决定机制研究进行了综述,主要包括性别决定基因概况和重要性别决定相关基因的分子级联互作关系。目前发现昆虫重要性别决定相关基因主要集中在常染色体上,且部分基因之间存在紧密的级联互作,如Sxl,tra,dsx,csd和fem等。在这些基因中,tra/fem→dsx的调控模式在已报道的昆虫中存在共性,即tra和dsx相对较保守且tra通过性特异剪切来调控下游dsx的转录形式。目前大多数昆虫的性别决定机制还不清楚,但近年来模式昆虫性别决定机制取得了一定进展,对非模式昆虫的研究还处于起步阶段但却越来越受到重视。     

PCR 扩增Sry 基因进行鲸类动物性别的鉴定

哺乳动物Y染色体短臂上的Sry 基因决定雄性发育方向。本研究参照哺乳动物Sry 基因保守区序列设计引物, 以非性别特异性的线粒体DNA 细胞色素b 基因作为阳性对照, 用PCR 扩增江豚、长喙真海豚等鲸类动物的Sry 基因片断并对其进行凝胶电泳分析来鉴定鲸类动物的性别。通过此方法对87 个已知性别鲸类动物标本的检验, 结果完全正确, 并进一步应用此方法成功地完成了另外33 个未知性别鲸类标本的性别鉴定。由此建立了一套简单、快速、可靠的鲸类动物的性别鉴定方法。     

棕色田鼠不同性别中R-spondin1基因的特殊表达

哺乳动物睾丸决定的诱导一般依赖于Sry基因,然而棕色田鼠指名亚种的Sry基因已经丢失,而棕色田鼠指名亚种雌雄个体依然有繁殖能力。我们在研究涉及性别决定的一些基因时,发现R-spondin1与性别决定有关。为了探讨R-spondin1在棕色田鼠中的性别决定中的作用,我们用RT-PCR检测R-spondin1在棕色田鼠性腺中的表达。研究结果表明R-spondin1仅在出生后不久的棕色田鼠指名亚种雌性个体的卵巢中表达,在出生后的棕色田鼠指名亚种雄性个体睾丸中未见其表达,这说明R-spondin1可能在棕色田鼠指名亚种卵巢发育中具有某种角色。     

鸡性别决定机制及相关基因研究进展

鸡性别决定虽然同哺乳动物一样受遗传控制,但其性染色体组成为ZZ／ZW,同哺乳动物相反呈现雌异型,并且鸡性腺性别分化同一些低等脊椎动物一样易受性激素影响。目前参照哺乳动物性别决定相关基因已获得了一些鸡同源基因序列(AMH,SF1,DAX1,SOX9)和3个可能与鸡性别决定有重要关联的候选基因(DMRT1,ASW和FET1)。对这些基因的表达模式及其在层次调控中的功能比较分析结果显示,鸡性别决定的遗传机制同其它脊椎动物相对一致,但也有明显的不同。     

siRNA介导Sry基因沉默对小鼠胚胎性别决定基因表达的影响

为研究Sry基因的调控网络, 采用siRNA表达载体介导的RNAi技术, 特异性地抑制睾丸决定因子Sry在小鼠胚胎中的表达, 并观察Sry基因沉默后对在两性性腺分化中起重要作用的Wt1, Sf1, Dax1, Gata4, Sox9及Amh基因表达的影响。利用本课题组先前构建的siRNA重组表达载体(pSilencer4.1/Sry217及pSilencer4.1/Sry565), 通过尾静脉注射法导入妊娠9.5天(9.5 dpc)的母鼠体内, 在11.5 dpc时取胚胎, 对性别鉴定为雄性的胚胎以RT-PCR法和Western-blot检测Sry基因的表达抑制效果, 并同时用定量PCR法检测Wt1等上述性别决定相关基因表达变化情况。结果表明, 注射干扰质粒后48 h Sry基因的mRNA和蛋白表达水平均降低, 其中siRNA表达质粒pSilencer 4.1/Sry 565的抑制效果显著, 可达到80%的抑制率。Sry基因沉默后, Wt1基因表达量显著升高; Sf1, Dax1, Gata4, Sox9基因表达水平没有明显变化; Amh基因无表达。试验结果表明, Sry基因表达抑制会导致Wt1基因表达升高; 另外, Sry基因激活Sox9基因的表达可能需要其他的辅助因子协同作用。     

Sry基因的研究进展

性别决定基因（Sex region of Y chromosome, 人类以SRY,小鼠以Sry表示）的研究进展是近几年来人类在性别决定,性别分化研究中获得的最大的突破性成果,该文从SRY（Sry)发现前关于性别决定因子的研究,SRY（Sry)的确定,小鼠Sry的结构研究,小鼠Sry的表达研究及Sry下游基因的确定等5个方面对小鼠Sry的研究进展进行综述,对进一步深入研究Sry下游基因存在的瓶颈问题人了一定的分析,并提出核移植技术可能对研究Sry的调节及其下游基因所需的特殊实验材料展现了新的希望。     

Absence of the candidate male sex-determining gene dmrt1b(Y) of medaka from other fish species

Although the sex-determining genes are known in mammals, Drosophila, and C. elegans, little is known in other animals. Fishes are an attractive group of organisms for studying the evolution of sex determination because they show an amazing variety of mechanisms, ranging from environmental sex determination and different forms of hermaphroditism to classical sex chromosomal XX/XY or WZ/ZZ systems and modifications thereof. In the fish medaka, dmrt1b(Y) has recently been found to be the candidate male sex-determining gene. It is a duplicate of the autosomal dmrt1a gene, a gene acting in the sex determination/differentiation cascade of flies, worms, and mammals. Because in birds dmrt1 is located on the Z-chromosome, both findings led to the suggestion that dmrt1b(Y) is a "non-mammalian Sry" with an even more widespread distribution. However, although Sry was found to be the male sex-determining gene in the mouse and some other mammalian species, in some it is absent and has obviously been replaced by other genes that now fulfil the same function. We have asked if the same might be true of the dmrt1b(Y) gene. We find that the gene duplication generating dmrt1b(Y) occurred recently during the evolution of the genus Oryzias. The gene is absent from all other fish species studied. Therefore, it may not be the male-sex determining gene in all fishes.     

Sex chromosome evolution in non-mammalian vertebrates

Birds, reptiles, amphibia and fish have an enormous variety of chromosomal sex determination mechanisms that apparently do not follow any phylogenetic or taxonomic scheme. A similar picture is now emerging at the molecular level. Most genes that function downstream of the mammalian master sex-determining gene, Sry, have been found in non-mammalian vertebrates. Although the components of the machinery that determines sex seem to be conserved, their interaction and most importantly the initial trigger is not the same in all vertebrates. This variety is the consequence of the extremely dynamic process of the evolution of sex determination mechanisms and sex chromosomes, which is prone to create differences rather than uniformity.     

Cloning and mutational analysis of SRY.

A candidate for the male sex-determining gene has recently been isolated. This sex-determining gene (SRY) has been found to be mutated in some individuals with failed testis development, and, in mouse transgenesis, the SRY murine homologue (Sry) causes female-to-male sex reversal. The cloning of SRY should facilitate the characterisation of other genes in the testis-determining pathway and provide information on the mechanism of mammalian developmental decisions.     

Mammalian sex reversal and intersexuality: deciphering the sex-determination cascade

The sex-determination cascade constitutes a model of the exquisite mechanisms of gene regulation that lead to the development of mammalian embryos. The discovery of the sex-determining region of the Y chromosome (SRY) in the early 1990s was the first crucial step towards a general understanding of sex determination. Since then, several genes that encode proteins with a role in this cascade, such as WT1, SF-1, SOX9, DAX-1 and WNT4, have been identified. Many of the interactions between these proteins have still to be elucidated, while, no-doubt, others are still to be identified. The study of mammalian intersexes forms a promising way towards the identification of the still-missing genes and a comprehensive view of mammalian sex determination. Intersexuality in the goat, studied for over a century, will, presumably, bring to light new genes involved in the female sex-determination pathway.     

Characterisation and expression of Sox9 in the Leopard gecko, Eublepharis macularius

Since the discovery of the sex-determining gene, Sry, a number of genes have been identified which are involved in sex determination and gonadogenesis in mammals. Although Sry is known to be the testis-determining factor in mammals, this is not the case in non-mammalian vertebrates. Sox9 is another gene that has been shown to have a male-specific role in sex determination, but, unlike Sry, Sox9 has been shown to be involved in sex determination in mammals, birds, and reptiles. This is the first gene to be described that has a conserved role in sex determination in species with either chromosomal or environmental sex-determining mechanisms. Many reptiles do not have sex chromosomes but exhibit temperature-dependent sex determination (TSD). Sox9 has been shown to be expressed in both turtle and alligator during gonadogenesis. To determine if Sox9 also has a role in a gecko species with TSD, we studied gonadal expression of Sox9 during embryonic development of the Leopard gecko (Eublepharis macularius). Gecko Sox9 was found to be highly conserved at the nucleotide level when compared to other vertebrate species including human, chick, alligator, and turtle. Sox9 was found to be expressed in embryos incubated at the male-producing temperature (32.5 degrees C) as well as in embryos incubated at the female-producing temperatures (26 and 34 degrees C), Northern blot analysis showed that Sox9 was expressed at both temperatures from morphological stages 31 to 37. mRNA in situ hybridisation on isolated urogenital systems showed expression at both female- and male-producing temperatures up to stage 36. After this stage, no expression was seen in the female gonads but expression remained in the male. These data provide further evidence that Sox9 is an essential component of a testis-determining pathway that is conserved in species with differing sex-determining mechanisms.     

An Immune-Related Gene Evolved into the Master Sex-Determining Gene in Rainbow Trout, Oncorhynchus mykiss

Since the discovery of Sry in mammals [1, 2], few other master sex-determining genes have been identified in vertebrates [3-7]. To date, all of these genes have been characterized as well-known factors in the sex differentiation pathway, suggesting that the same subset of genes have been repeatedly and independently selected throughout evolution as master sex determinants [8, 9]. Here, we characterized in rainbow trout an unknown gene expressed only in the testis, with a predominant expression during testicular differentiation. This gene is a male-specific genomic sequence that is colocalized along with the sex-determining locus. This gene, named sdY for sexually dimorphic on the Y?chromosome, encodes a protein that displays similarity to the C-terminal domain of interferon regulatory factor 9. The targeted inactivation of sdY in males using zinc-finger nuclease induces ovarian differentiation, and the overexpression of sdY in females using additive transgenesis induces testicular differentiation. Together, these results demonstrate that sdY is a novel vertebrate master sex-determining gene not related to any known sex-differentiating gene. These findings highlight an unexpected evolutionary plasticity in vertebrate sex determination through the demonstration that master sex determinants can arise from the de novo evolution of genes that have not been previously implicated in sex differentiation.     

Evolutionary origin of the medaka Y chromosome

Genetic sex determination in an XX-XY chromosome system can be realized through a locus on the Y chromosome that makes the undifferentiated gonad develop into a testis. Although this mechanism is widespread, only in two cases so far have the corresponding master male sex-determining genes been identified. One is Sry, which initiates testes determination in most mammals. The other is dmrt1bY (syn. dmy), from the fish medaka, Oryzias latipes. The mammalian Y is roughly estimated to be over 200 million years old. The medaka Y may be considerably younger. A comparative analysis of the genus Oryzias revealed that one sister species of the medaka has dmrt1bY on a homologous Y chromosome, whereas in another closely related species only a non-sex-linked pseudogene is present. In all other species, dmrt1bY was not detected. The divergence time for the different species was determined with mitochondrial DNA sequences. The timing was confirmed by independent calculations based on dmrt1 sequences. We show that the medaka sex-determining gene originated approximately 10 million years ago. This makes dmrt1bY and the corresponding Y chromosome the youngest male sex-determining system, at least in vertebrates, known so far.