激活素受体相互作用蛋白1,2在小鼠脑神经细胞中的共表达

Co-expression of activin receptor-interacting proteins 1 and 2 in nerve cells of mouse brain

目的探讨激活素受体相互作用蛋白1(activin receptor-interaction protein 1,ARIP1)和ARIP2在小鼠脑神经细胞中的共表达。方法 RT-PCR法检测C57BL小鼠小脑、垂体、大脑、脾脏、心脏、肾脏、肝脏、胰腺组织中ARIP1和ARIP2基因mRNA的转录;以融合蛋白GST-ARIP1C和MBP-ARIP2C作为抗原免疫新西兰白兔,制备抗ARIP1和ARIP2抗体,经A蛋白亲和层析柱纯化后,ELISA法检测抗体的交叉反应;用制备的抗体,采用免疫组织化学染色检测ARIP1和ARIP2在小鼠脑组织中的表达;RT-PCR法检测小鼠神经母细胞瘤Neuro-2a细胞中激活素ⅡA型受体(activin receptorⅡA,ActRⅡA)、ARIP1和ARIP2基因mRNA的转录。结果在小鼠大脑、小脑及垂体组织中可见ARIP1基因mRNA的转录,在小鼠各部位组织中均可见ARIP2基因mRNA的转录;制备的兔抗ARIP1和ARIP2抗体无交叉反应;ARIP1和ARIP2在小鼠脑组织的海马和下丘脑同时表达;ActRIⅡA及ARIP1和ARIP2 mRNA在神经元样细胞系Neuro-2a细胞中共表达。结论 ARIP1和ARIP2可在小鼠脑神经细胞中共表达。

Objective To investigate the co-expression of activin receptor-interacting proteins 1 and 2（ARIP1,2） in nerve cells of mouse brain.Methods The transcription levels of ARIP1 and ARIP2 mRNAs in cerebellum,pituitary gland,brain,spleen,heart,kidney,liver and pancreas were determined by RT-PCR.Antibodies against ARIP1 and ARIP2 were prepared by immunizing New Zealand rabbits with fusion protein GST-ARIP1C and MBP-ARIP2C respectively,purified by protein A affinity chromatography and determined for cross reaction by ELISA.The expressions of ARIP1 and ARIP2 in mouse brain tissue were determined by immunohistochemical staining with the prepared antibodies.The transcriptions of activin receptor ⅡA（ActRⅡA）,ARIP1 and ARIP2 mRNAs in mouse neuroblastoma（NB） Neuro-2a cells were determined by RT-PCR.Results The transcription of ARIP1 mRNA was observed in brain,cerebellum and pituitary gland of mice,while that of ARIP2 mRNA in various tissues.The prepared rabbit antibodies against ARIP1 and ARIP2 showed no cross reaction.ARIP1 and ARIP2 were expressed simultaneously in hippocampus and hypothalamus of mice.ActRⅡA,ARIP1 and ARIP2 were co-expressed in Neuro-2a cells.Conclusion ARIP1 and ARIP2 were co-expressed in nerve cells of mouse brain.