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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 coexpression of activin receptor-interaction protein 1 (ARIP1) and ARIP2 in mouse brain neurons. Methods The mRNA transcription of ARIP1 and ARIP2 genes in cerebellum, pituitary, brain, spleen, heart, kidney, liver and pancreas of C57BL mice was detected by RT-PCR. New Zealand white rabbits were immunized with the fusion proteins GST-ARIP1C and MBP-ARIP2C The anti-ARIP1 and ARIP2 antibodies were prepared and purified by Protein A affinity chromatography. The antibody cross-reactivity was detected by ELISA. The expression of ARIP1 and ARIP2 in mouse brain was detected by immunohistochemistry. The transcription of activin receptor Ⅱ A (ActRⅡA), ARIP1 and ARIP2 mRNA in mouse neuroblastoma Neuro-2a cells was detected by RT-PCR. Results ARIP1 gene mRNA transcription was observed in mouse cerebellum, cerebellum and pituitary tissues. ARIP2 gene mRNA transcription was observed in all tissues of mice. No cross-reaction was observed between rabbit anti-ARIP1 and ARIP2 antibodies. ARIP1 and ARIP2 were small ActinⅡA and ARIP1 and ARIP2 mRNA were co-expressed in neuronal-like cell line Neuro-2a cells. Conclusion ARIP1 and ARIP2 can be co-expressed in mouse brain neurons.