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罗非鱼前体促黑素细胞激素原信使核糖核酸在下丘脑和神经垂体细胞中的表达。

Expression of tilapia prepro-melanin-concentrating hormone mRNA in hypothalamic and neurohypophysial cells.

作者信息

Gröneveld D, Eckhardt E R, Coenen A J, Martens G J, Balm P H, Wendelaar Bonga S E

机构信息

Department of Animal Physiology, Faculty of Science, University of Nijmegen, The Netherlands.

出版信息

J Mol Endocrinol. 1995 Apr;14(2):199-207. doi: 10.1677/jme.0.0140199.

Abstract

Melanin-concentrating hormone (MCH) is a neuropeptide involved in background adaptation in teleost fish, and in multiple regulatory functions in mammals and fish. To study the expression of the MCH preprohormone (ppMCH) in teleosts, we first cloned a hypothalamic cDNA encoding the complete ppMCH of tilapia (Oreochromis mossambicus), and a cRNA probe derived from a 270 bp ppMCH cDNA fragment was used for the expression studies. The level of ppMCH mRNA expression in tilapia hypothalamus, measured by dot blot analysis, was significantly higher in fish adapted to a white background than in black-adapted animals, which is in accordance with the reported MCH plasma and tissue concentrations in fish. Northern blot analysis not only revealed a strong ppMCH mRNA signal in the hypothalamus, but also the presence of ppMCH mRNA in the neurointermediate lobe (NIL) of the pituitary. In situ hybridization and immunocytochemistry showed that ppMCH mRNA as well as MCH immunoreactivity are located in perikarya of two hypothalamic regions, namely in the nucleus lateralis tuberis (NLT) and the nucleus recessus lateralis (NRL). Quantitative analysis by dot blot hybridization revealed about eight times more ppMCH mRNA in the NLT than in the NRL and NIL of mature tilapias. ppMCH mRNA in the NIL could be localized to cell bodies of the neurohypophysis, which were also MCH immunoreactive.

摘要

黑色素浓缩激素(MCH)是一种神经肽,参与硬骨鱼的背景适应以及哺乳动物和鱼类的多种调节功能。为了研究MCH前激素原(ppMCH)在硬骨鱼中的表达,我们首先克隆了罗非鱼(莫桑比克罗非鱼)下丘脑编码完整ppMCH的cDNA,并使用源自270 bp ppMCH cDNA片段的cRNA探针进行表达研究。通过斑点印迹分析测量,适应白色背景的罗非鱼下丘脑ppMCH mRNA表达水平显著高于适应黑色背景的动物,这与报道的鱼类MCH血浆和组织浓度一致。Northern印迹分析不仅在下丘脑中揭示了强烈的ppMCH mRNA信号,还显示垂体神经中间叶(NIL)中存在ppMCH mRNA。原位杂交和免疫细胞化学表明,ppMCH mRNA以及MCH免疫反应性位于下丘脑两个区域的神经元胞体中,即外侧结节核(NLT)和外侧隐窝核(NRL)。通过斑点印迹杂交进行的定量分析显示,成熟罗非鱼的NLT中ppMCH mRNA比NRL和NIL中的多约八倍。NIL中的ppMCH mRNA可定位于神经垂体的细胞体,这些细胞体也具有MCH免疫反应性。

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