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生长激素家族成员基因敲低对斑马鱼发育的影响。

The effects of the members of growth hormone family knockdown in zebrafish development.

作者信息

Zhu Yong, Song Danyin, Tran Ngoc-Tuyen, Nguyen Nhu

机构信息

Department of Biology, East Carolina University, Howell Science Complex, 1000 E. 5th Street, Greenville, NC 27858-4553, USA.

出版信息

Gen Comp Endocrinol. 2007 Feb;150(3):395-404. doi: 10.1016/j.ygcen.2006.10.009. Epub 2006 Dec 1.

Abstract

Growth hormone (GH), prolactin (PRL), and somatolactin (SL) are members of GH/PRL superfamily. These hormones are involved in the regulation of an array of physiological processes, including growth, lactation, and osmoregulation. While recent evidence has shown the GH, PRL, and SL gene transcripts and protein products are expressed during early zebrafish development, their functions at this time of embryogenesis remain unknown. In the current study, antisense morpholino oligonucleotide (MO) inhibition of gh, prl, and sl gene translation was used to examine the effects of gene knockdown on hormone function in zebrafish development. We observed that PRL, SLalpha and SLbeta MO treatment all affected development. PRL MO-treated embryos showed defects in gas bladder inflation, reduced head and eye size, shorter body length and fewer melanophores than untreated controls, whereas SLalpha and SLbeta MO-treated embryos were only defective in gas bladder inflation, GH MO-inhibition of GH specific translation did not lead to any discernable morphological changes within 10 days post fertilization (dpf). The effects of PRL knockdown were further verified using a second PRL morpholino antisense and by a rescue experiment with in vitro transcribed prl mRNA containing 5 nucleotide mismatch within the PRL-MO binding region. These results provide the first evidence that members of the GH/PRL superfamily play a role in proper development of various structures including the head, eyes, melanophores and the gas bladder in zebrafish.

摘要

生长激素(GH)、催乳素(PRL)和生长抑素(SL)是GH/PRL超家族的成员。这些激素参与一系列生理过程的调节,包括生长、泌乳和渗透压调节。虽然最近的证据表明GH、PRL和SL基因转录本及蛋白质产物在斑马鱼早期发育过程中表达,但其在胚胎发生这个阶段的功能仍不清楚。在本研究中,利用反义吗啉代寡核苷酸(MO)抑制gh、prl和sl基因的翻译,以检测基因敲低对斑马鱼发育中激素功能的影响。我们观察到,PRL、SLα和SLβ的MO处理均影响发育。与未处理的对照组相比,经PRL MO处理的胚胎在鳔充气方面存在缺陷,头部和眼睛尺寸减小,体长缩短,黑素细胞减少,而经SLα和SLβ MO处理的胚胎仅在鳔充气方面存在缺陷,GH MO抑制GH特异性翻译在受精后10天内(dpf)未导致任何可察觉的形态变化。使用第二种PRL吗啉代反义寡核苷酸以及通过在PRL-MO结合区域内含有5个核苷酸错配的体外转录prl mRNA进行拯救实验,进一步验证了PRL敲低的效果。这些结果首次证明,GH/PRL超家族成员在斑马鱼包括头部、眼睛、黑素细胞和鳔在内的各种结构的正常发育中发挥作用。

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