草鱼成纤维细胞gcngr1基因的表达分析

Expression analysis of gcngr1 gene in grass carp(Ctenopharyngodon idellus) PSF cells

Ngr1(nogo-66 receptor)是在哺乳类上发现的一种神经元受体,可调节轴突的可塑性并抑制损伤中枢神经系统(CNS)的再生,最新研究还发现它是哺乳动物呼肠孤病毒(mammalian reovirus)的神经受体。草鱼呼肠孤病毒(grass carp reovirus,GCRV)可引发草鱼(Ctenopharyngodon idellus)出血病导致高死亡率,开展GCRV受体相关研究可有助于了解病毒的致病机制。该研究在草鱼吻端成纤维细胞(PSF)中克隆到草鱼ngr1 c DNA序列(下文简称为gcngr1),发现其与哺乳动物呼肠孤病毒神经受体基因Ngr1有相似的结构序列;采用q RT-PCR方法检测该基因在PSF细胞的表达情况,结果显示受草鱼呼肠孤病毒(GCRV-GD108株)感染后gcngr1 m RNA的表达量显著上升,与病毒的增殖趋势基本一致;病毒经病毒抗体孵育后再感染PSF细胞,细胞中病毒的增殖水平下降,gcngr1m RNA的表达量也显著下降。该研究结果提示gcngr1与病毒的感染相关,为进一步分析gcngr1是否为GCRV神经元受体提供依据。

The Nogo-66 receptor （ngrl）, expresses in neuron surface, can inhibit the regeneration of injury central nervous system （CNS） and regulate axon plasticity in mammals. Recent studies have shown that it also serves as a mammalian reovirus （MRV） neu- ron receptor. Grass carp reovirus （GCRV） can invade grass carp （Ctenopharyngodon idellus） and cause high mortality, so it would be helpful to understand the mechanisms of pathogenicityis on GCRV receptor. In this study, the grass carp ngrl （gcngrl） cDNA was cloned from grass carp snout fibroblasts cells （PSF） , which shared similar sequence with MRV receptor Ngrl ; qRT-PCR was per- formed to detect the expression of gcngrl and virus proliferation in grass carp snout fibroblasts （PSF） cells challenged by GCRV- GDI08. The gcngrl expression in PSF cells showed a significant increase after being infected by GCRV-GDI08, which was consistent with the trend of virus reproduction. In addition, GCRV-GD108 was incubated with polyclonal antibody of GCRV-GDI08 firstly and then infected PSF cells. A significant decrease in virus reproduction was observed, and gcngrl mRNA expression also decreased in PSF cells. The results show that the expression of gcngrl is related with GCRV infection in PSF cells, which provides references for determining whether gcngrl is a GCRV receptor or not.