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目的 :采用同位素相对标记和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQi)技术分析耳蜗核(cochlear nucleus,CN)区域特异性膜蛋白。方法 :出生60 d的雄性SD大鼠分为4组,分别是CN、下丘(inferior colliculus,IC)、上橄榄核(superior oliver complex,SOC)、大脑其余部位(Rest)。提取这4个部位组织的细胞膜蛋白,再采用iTRAQi技术检测出CN区域特异性蛋白。通过Uniprot检索这些蛋白的功能和参与的生物代谢过程。结果:本实验最终得到17种CN区域特异性膜蛋白。对CN区域蛋白进行基因本体(gene ontology,GO)分析发现,这些蛋白主要参与生物调节与发育。UNIPROT检索发现3种蛋白与神经系统功能相关,即半乳糖凝集素-1(galectin-1,Gal-1)、膜联蛋白A2(annexin-A2,AXNA2)和外周蛋白(peripherin,PRPH)。Gal-1与神经生长发育相关;AXNA2参与多种生物调节过程;PRPH与神经退行性病变相关。其中,Gal-1可促进轴突生长、调节突触可塑性且具有神经保护作用;AXNA2可能参与了神经递质释放过程,且有神经营养作用;PRPH在高表达时具有神经毒性,和神经丝蛋白共同参与神经退行性病变。结论:Gal-1、ANXA2和PRPH在耳蜗核中高度表达,可能与中枢信息处理相关,这为进一步研究中枢听觉处理障碍发病机制提供了一定的理论基础。
OBJECTIVE: To analyze the region-specific membrane proteins of cochlear nucleus (CN) using isobaric tags for relative and absolute quantitation (iTRAQi). Methods: Sixty male SD rats were divided into 4 groups: CN, inferior colliculus (IC), superior oliver complex (SOC) and the rest of the brain. The cell membrane proteins of these four tissues were extracted and the CN region-specific proteins were detected by iTRAQi technique. Retrieval of these proteins by Uniprot functions and participates in the biological metabolism process. Results: Seventeen CN region-specific membrane proteins were finally obtained in this experiment. Gene ontology (GO) analysis of CN region proteins revealed that these proteins are mainly involved in biological regulation and development. UNIPROT search found that the three proteins were related to nervous system function, namely galectin-1 (Gal-1), annexin-A2 (AXNA2) and peripherin (PRPH). Gal-1 is involved in nerve growth and development; AXNA2 is involved in many biological processes; PRPH is associated with neurodegenerative diseases. Among them, Gal-1 can promote axonal growth, regulate synaptic plasticity and have neuroprotective effects; AXNA2 may be involved in the process of neurotransmitter release, and have neurotrophic effects; PRPH is neurotoxic when high expression, and neurofilament Together to participate in neurodegenerative diseases. CONCLUSIONS: Gal-1, ANXA2 and PRPH are highly expressed in the cochlear nucleus and may be related to central information processing. This provides a theoretical basis for further study on the pathogenesis of central auditory processing disorders.