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Differential expression of fertility genes boule and dazl in Chinese sturgeon (Acipenser sinensis), a basal fish

机译:基底鱼类中华st(Acipenser sinensis)的育性基因boule和dazl的差异表达

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The gene family DAZ (deleted in Azoospermia), including boule, dazl and DAZ, performs highly conserved functions in germ cell development and fertility across animal phyla. Differential expression patterns have been demonstrated for the family members in invertebrates and vertebrates including fish. Here, we report the identification of boule and dazl and their expression at both RNA and protein levels in developing and mature gonads of Chinese sturgeon (Acipenser sinensis). Firstly, the isolation of the boule and dazl genes in Chinese sturgeon and the observation of the two genes in coelacanth suggest that dazl originated after the divergence of bony fish from cartilaginous fish but before the emergence of the Actinistia. Quantitative real-time PCR and western blot analyses reveal that boule and dazl RNA and proteins are restricted to the testis and ovary. In situ hybridization and fluorescent immunohistochemistry show that the bisexual mitotic and meiotic germ cell expression of dazl RNA and protein is conserved in vertebrates, while Chinese sturgeon boule RNA and protein exhibit mitotic and meiotic expression in the testis, and also likely display mitotic and meiotic expression in female. Moreover, we directly demonstrate for the first time that sturgeon Balbiani body/mitochondrial cloud disperses in the cytoplasm of early developing oocytes and co-localizes with Dazl to some extent. Finally, urbilaterian boule may also have an ancestral function in oogenesis. Taken together, these results provide useful information on the evolution of DAZ family genes, expression patterns and functions in animal reproduction.
机译:DAZ基因家族(在无精子症中缺失),包括短毛,dazl和DAZ,在整个动物门的生殖细胞发育和繁殖力中具有高度保守的功能。在无脊椎动物和包括鱼的脊椎动物中,已经证明了其家族成员的差异表达模式。在这里,我们报道了中华st(Acipenser sinensis)发育中和成熟的性腺中的小球和dazl的鉴定及其在RNA和蛋白质水平上的表达。首先,中华Chinese中的boul基因和dazl基因的分离以及腔棘鱼中两个基因的观察表明,dazl起源于骨质鱼类与软骨鱼类的分叉,但在放线Act出现之前。实时定量PCR和Western印迹分析表明,小球和dazl RNA和蛋白质仅限于睾丸和卵巢。原位杂交和荧光免疫组化显示,脊椎动物中dazl RNA和蛋白质的双性有丝分裂和减数分裂生殖细胞表达是保守的,而中华st的RNA和蛋白质在睾丸中表现出有丝分裂和减数分裂表达,还可能显示有丝分裂和减数分裂表达。在女性。此外,我们首次直接证明st鱼Balbiani体/线粒体云分散在早期发育的卵母细胞的细胞质中,并在一定程度上与Dazl共定位。最后,脐带小卵在卵子发生中也可能具有祖先的功能。综上所述,这些结果提供了有关DAZ家族基因的进化,表达模式和动物繁殖功能的有用信息。

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