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石斑鱼促甲状腺激素β亚基启动子驱动的转基因斑马鱼标记近端肾小管发育。

Grouper tshβ promoter-driven transgenic zebrafish marks proximal kidney tubule development.

作者信息

Wang Yang, Sun Zhi-Hui, Zhou Li, Li Zhi, Gui Jian-Fang

机构信息

State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Wuhan, China.

出版信息

PLoS One. 2014 Jun 6;9(6):e97806. doi: 10.1371/journal.pone.0097806. eCollection 2014.

Abstract

Kidney tubule plays a critical role in recovering or secreting solutes, but the detailed morphogenesis remains unclear. Our previous studies have found that grouper tshβ (gtshβ) is also expressed in kidney, however, the distribution significance is still unknown. To understand the gtshβ role and kidney tubule morphogenesis, here, we have generated a transgenic zebrafish line Tg(gtshβ:GFP) with green fluorescent protein driven by the gtshβ promoter. Similar to the endogenous tshβ in zebrafish or in grouper, the gtshβ promoter-driven GFP is expressed in pituitary and kidney, and the developing details of proximal kidney tubule are marked in the transgenic zebrafish line. The gfp initially transcribes at 16 hours post fertilization (hpf) above the dorsal mesentery, and partially co-localizes with pronephric tubular markers slc20a1a and cdh17. Significantly, the GFP specifically localizes in proximal pronephric segments during embryogenesis and resides at kidney duct epithelium in adult fish. To test whether the gtshβ promoter-driven GFP may serve as a readout signal of the tubular development, we have treated the embryos with retinoic acid signaing (RA) reagents, in which exogenous RA addition results in a distal extension of the proximal segments, while RA inhibition induces a weakness and shortness of the proximal segments. Therefore, this transgenic line provides a useful tool for genetic or chemical analysis of kidney tubule.

摘要

肾小管在溶质的重吸收或分泌中起着关键作用,但其详细的形态发生过程仍不清楚。我们之前的研究发现,石斑鱼促甲状腺激素β(gtshβ)在肾脏中也有表达,然而,其分布意义仍不清楚。为了了解gtshβ的作用和肾小管的形态发生,在此,我们构建了一个转基因斑马鱼品系Tg(gtshβ:GFP),其中绿色荧光蛋白由gtshβ启动子驱动。与斑马鱼或石斑鱼中的内源性促甲状腺激素β相似,gtshβ启动子驱动的绿色荧光蛋白在垂体和肾脏中表达,并且在转基因斑马鱼品系中标记了近端肾小管的发育细节。绿色荧光蛋白最初在受精后16小时(hpf)在背系膜上方转录,并与前肾管标记物slc20a1a和cdh17部分共定位。值得注意的是,绿色荧光蛋白在胚胎发育过程中特异性定位于近端前肾段,在成鱼中位于肾管上皮。为了测试gtshβ启动子驱动的绿色荧光蛋白是否可以作为肾小管发育的读出信号,我们用视黄酸信号(RA)试剂处理胚胎,其中添加外源性RA会导致近端段向远端延伸,而抑制RA则会导致近端段变弱和变短。因此,这个转基因品系为肾小管的遗传或化学分析提供了一个有用的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b88b/4048157/ef1864764cb1/pone.0097806.g001.jpg

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