Establishment of transgenic zebrafish (Danio rerio) models expressing fluorescence proteins in the oocytes and somatic supporting cells
Establishment of transgenic zebrafish (Danio rerio) models expressing fluorescence proteins in the oocytes and somatic supporting cells
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建立在卵母细胞和体细胞支持细胞中表达荧光蛋白的转基因斑马鱼（Danio rerio）模型
DOI：
10.1016/j.ygcen.2021.113907
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发表时间：
2021
影响因子：
2.7
通讯作者：
Wei Ge
中科院分区：
文献类型：
--
作者：
Yanlong Song;Wei Hu;Wei Ge
性腺支持细胞和生殖细胞之间的相互作用在性腺发育过程中起着至关重要的作用。在FISH中，这一过程涉及多种局部生长因子，如生长分化因子9(GDF9)和性腺体细胞衍生因子(GSDF)，它们都是转化生长因子-(TGFR-)超家族成员。GDF9是一种卵母细胞分泌因子，对哺乳动物和鱼类的卵泡发生都有很强的调节作用。相反，GSDF由支持生殖细胞发育的性腺体细胞(即睾丸中的支持细胞和卵巢中的颗粒细胞)表达。在本研究中，我们建立了两个转基因斑马鱼模型，并证明GDF9的2.7kb近端启动子区域驱动mCherry在卵母细胞中的特异性表达，而2.1kb的GSDF近端启动子区域驱动绿色荧光蛋白(EGFP)在支持细胞和颗粒细胞中的表达增强。这些近端启动子包含了足够的信息来分别模拟内源GDF9和GSDF在斑马鱼中的时空表达模式。在TG(GDF9：mCherry)鱼中，mCherry在初级生长阶段的卵母细胞中表达较弱，而在进入次级生长阶段的卵母细胞中表达较强。在TG(GSDF：EGFP)FISH中，EGFP阳性的支持细胞分布在睾丸生精囊周围，而EGFP阳性的颗粒细胞位于卵巢卵泡层的外侧。EGFP阳性的支持细胞和颗粒细胞似乎起源于性腺背侧上皮。TG(GDF9：mCherry)和TG(GSDF：EGFP)斑马鱼模型适合于研究性腺的发育和功能，特别是生殖细胞和支持体细胞之间的相互作用。
The interaction between gonadal somatic support cells and germ cells plays a crucial role in gonadal development. In fish, the process involves various local growth factors such as growth differentiation factor 9 (Gdf9) and gonadal soma-derived factor (Gsdf), which are both members of the transforming growth factor- (TGF-) superfamily. Gdf9, an oocyte-secreted factor, is a potent regulator of folliculogenesis in both mammals and fish. By contrast, Gsdf is expressed by the gonadal somatic cells (i.e., Sertoli cells in the testis and granulosa cells in the ovary) that support germ cell development. In this study, we established two transgenic zebrafish models, and demonstrated that the 2.7-kb proximal promoter region of gdf9 drove mCherry expression specifically in the oocytes, whereas the 2.1-kb proximal promoter region of gsdf drove enhanced green fluorescent protein (eGFP) expression in the Sertoli cells and granulosa cells. These proximal promoters contained sufficient information to respectively mimic the spatiotemporal expression patterns of endogenous gdf9 and gsdf in zebrafish. In the Tg(gdf9:mCherry) fish, mCherry was weakly expressed in the oocytes at primary growth stage but strongly expressed in those entering the secondary growth phase. In the Tg(gsdf:eGFP) fish, eGFP-positive Sertoli cells were distributed around spermatogenic cysts in the testis, whereas eGFP-positive granulosa cells were located at the outer side of the follicle layer in the ovary. The eGFP-positive Sertoli cells and granulosa cells seemed to have originated from the dorsal epithelium of the gonads. These Tg(gdf9:mCherry) and Tg(gsdf:eGFP) zebrafish models are suitable for studying gonadal development and function especially on the interaction between germ cells and supporting somatic cells.