兔神经肽W及其受体NPWR1基因克隆及分布定位

摘要

[Objective] This study was carried out to obtain sequence of NPW and NPWR1 (GPR7) gene, and reveal the expression of NPW and NPWR1 mRNA in rabbit organs. [Method] The partial coding sequences of NPW and NPWR1 were obtained from rabbit brain using the approach of gene cloning. The expressions of NPW and NPWR1 mRNA in rabbit brain were studied by semi-quantitative RT-PCR and in situ hybridization. [Result] The length of NPW and NPWR1 was 234 bp and 508 bp, respectively. The sequences of the products were confirmed to be those of NPW and NPWR1 in rabbit and enrolled in GenBank (accession numbers: HQ596517 and HQ596518). Phylogenetic analysis showed that amino sequence of rabbit NPS precursor had high homology compared with other animals. The expression of NPW and NPWR1 mRNA was extensive in rabbit organs, with majority of NPWR1 mRNA in the hippocampus, cerebellum, hypothalamus and medulla oblongata in the central nervous system. [Conclusion J NPW and its receptor gene in rabbit were conservative, and the expression of NPW and NPWR1 mRNA in rabbit organs were extensive, suggesting that they are involved in regulations of various physiological functions.%[目的]研究神经肽W (NPW)及其受体1(NPWR1,GPR7) mRNA在兔各器官组织的表达情况.[方法]利用基因克隆方法克隆出兔神经肽W及其受体的cDNA序列片段,以GAPDH为内参基因,采用相对定量RT-PCR方法检测NPW及NPWR1 mRNA在兔体内各组织中的表达情况,用原位杂交方法研究NPWR1阳性细胞在兔脑的分布定位.[结果]克隆出兔NPW及其受体NPWR1的基因部分序列,NPW基因片段长234 bp,NPWR1基因片段长508 bp,GenBank登录号分别为HQ596517和HQ596518;经DNAStar软件分析,不同物种之间NPW及其受体基因序列的同源性较高.NPW及其受体在兔各组织广泛分布；NPWR1 mRNA在脑内主要分布于海马、小脑、下丘脑、延髓等部位.[结论]兔NPW及其受体基因在进化上是保守的;NPW及其受体在兔体内广泛表达,提示其参与多种生理功能的调节.