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甲状腺激素诱导及虹鳟自然发育过程中视网膜视蛋白基因表达的时空特征

Spatio-temporal characterization of retinal opsin gene expression during thyroid hormone-induced and natural development of rainbow trout.

作者信息

Veldhoen Kathy, Allison W Ted, Veldhoen Nik, Anholt Bradley R, Helbing Caren C, Hawryshyn Craig W

机构信息

Department of Biology, University of Victoria, Victoria, British Columbia, Canada.

出版信息

Vis Neurosci. 2006 Mar-Apr;23(2):169-79. doi: 10.1017/S0952523806232139.

Abstract

The abundance and spatial distribution of retinal cone photoreceptors change during thyroid hormone (TH)-induced and natural development of rainbow trout (Oncorhynchus mykiss). These changes are thought to allow the fish to adapt to different photic environments throughout its life history. To date, the ontogeny of rainbow trout cone photoreceptors has been examined using physiological and morphological approaches. In this study, we extended these observations by measuring opsin gene expression in retinal quadrants during natural and TH-induced development. Gene expression during natural development was investigated in retinae from fish at both parr and smolt stages. The role of TH in modulating opsin gene expression was determined in TH-treated parr and control fish sampled after two, nine, and 22 days of treatment. Total RNA was isolated from each retinal quadrant and steady-state opsin mRNA levels were measured using reverse transcriptase real-time quantitative polymerase chain reaction (QPCR) analysis. Expression of ultraviolet-sensitive opsin (SWS1), rod opsin (RH1), middle wavelength-sensitive opsin (RH2), and long wavelength-sensitive opsin (LWS) transcripts vary spatially in the parr retina. Smolts, compared to parr, had downregulated SWS1 expression in all quadrants, lower LWS expression dorsally, higher RH1 expression nasally, and higher RH2 expression dorsally. In TH-treated parr, SWS1 opsin expression was downregulated in the nasal quadrants by two days. SWS1 displayed the greatest degree of downregulation in all quadrants after nine days of treatment, with an increase in short wavelength-sensitive (SWS2) and RH2 opsin mRNA expression in the temporal quadrants. This study reveals that opsin genes display spatially significant differences within rainbow trout retina in their level of mRNA expression, and that regulation of opsin expression is a dynamic process that is influenced by TH. This is particularly evident for SWS1 gene expression in parr following TH-induced and natural development.

摘要

在甲状腺激素(TH)诱导的虹鳟(Oncorhynchus mykiss)发育过程以及自然发育过程中,视网膜视锥光感受器的丰度和空间分布会发生变化。这些变化被认为使鱼类能够在其整个生命历程中适应不同的光照环境。迄今为止,已使用生理学和形态学方法研究了虹鳟视锥光感受器的个体发育。在本研究中,我们通过测量自然发育和TH诱导发育过程中视网膜象限中的视蛋白基因表达来扩展这些观察结果。在幼鲑和稚鱼阶段的鱼类视网膜中研究了自然发育过程中的基因表达。在处理2天、9天和22天后对经TH处理的幼鲑和对照鱼进行采样,以确定TH在调节视蛋白基因表达中的作用。从每个视网膜象限中分离出总RNA,并使用逆转录实时定量聚合酶链反应(QPCR)分析来测量视蛋白mRNA的稳态水平。在幼鲑视网膜中,紫外线敏感视蛋白(SWS1)、视杆视蛋白(RH1)、中波长敏感视蛋白(RH2)和长波长敏感视蛋白(LWS)转录本的表达在空间上存在差异。与幼鲑相比,稚鱼在所有象限中SWS1表达下调,背侧LWS表达较低,鼻侧RH1表达较高,背侧RH2表达较高。在经TH处理的幼鲑中,鼻侧象限中的SWS1视蛋白表达在2天时下调。处理9天后,SWS1在所有象限中下调程度最大,颞侧象限中短波长敏感(SWS2)和RH2视蛋白mRNA表达增加。这项研究表明,视蛋白基因在虹鳟视网膜内mRNA表达水平上存在显著的空间差异,并且视蛋白表达的调节是一个受TH影响的动态过程。这在TH诱导发育和自然发育后的幼鲑中SWS1基因表达方面尤为明显。

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