GFPexpression pattern in pituitary and gonads under the control of nuclear progesterone receptor promoter in transgenic zebrafish
GFPexpression pattern in pituitary and gonads under the control of nuclear progesterone receptor promoter in transgenic zebrafish
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转基因斑马鱼核孕酮受体启动子控制下垂体和性腺中 GFP 的表达模式
DOI：
10.1002/dvdy.213
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发表时间：
2020-06-29
影响因子：
2.5
通讯作者：
Chen, Shi Xi
中科院分区：
文献类型：
--
作者：
Huang, Jing;Zhang, Ting Ting;Chen, Shi Xi
背景：核孕酮受体(PGR)是一种配体依赖的转录因子，主要负责介导孕酮在脊椎动物生殖中的作用。为了开发一种全面的方法来阐明PGR在生殖中的作用，需要关于PGR在生殖系统中表达的细胞定位的信息。结果获得了由pGR基因启动子序列驱动表达增强型绿色荧光蛋白(EGFP)的转基因斑马鱼Tg(PGR：EGFP)。EgfpmRNA的组织分布模式与pgrmRNAIntg(PGR：EGFP)的表达一致。在脑垂体部，GFP信号位于近远侧部。为了更好地识别GFP信号在性腺中的细胞定位，将Tg(PGR：EGFP)系与Tg(GSDF：NFSB-mCherry)系杂交，分别在颗粒细胞和卵巢和睾丸的Sertoli细胞中特异表达硝酸还原酶-mCherry融合蛋白。睾丸组织成像显示GFP仅表达于间质细胞。在卵巢中，GFP的表达与mCherry信号共存于颗粒细胞。有趣的是，我们还发现了一些靠近血管分支的非颗粒细胞，它们比颗粒细胞表达更强的GFP信号。结论分析Tg(PGR：EGFP)在斑马鱼中的表达为研究PGR在生殖中的作用提供了新的途径。
Background The nuclear progesterone receptor (Pgr) is a ligand-dependent transcription factor primarily responsible for mediating progesterone actions relevant for reproduction across vertebrates. Information on the cellular localization of Pgr expression in the reproductive system is required for developing a comprehensive approach to elucidate the role of Pgr in reproduction. Results We generated transgenic zebrafishTg(pgr:eGFP)that express enhanced green fluorescent protein (eGFP) driven by promoter sequence ofpgrgene. The tissue distribution pattern ofegfpmRNA is consistent with thepgrmRNA expression inTg(pgr:eGFP). In the pituitary, GFP signals are found in the proximal pars distalis. In order to better discern the cellular localization of GFP signals in gonads,Tg(pgr:eGFP)line was crossed withTg(gsdf:nfsB-mCherry)line, specifically expressing nitroreductase-mCherry fusion protein in granulosa and Sertoli cells in ovary and testis, respectively. Imaging of testis tissue showed that GFP expression was confined to Leydig cells. In the ovary, GFP expression colocalized with the mCherry signal in granulosa cells. Intriguingly, we also identified some non-granulosa cells close to where blood vessels branched, expressing stronger GFP signals than granulosa cells. Conclusions AnalyzingTg(pgr:eGFP)expression in zebrafish provided leads toward new routes to study the role of Pgr in reproduction.