Guo Ting-Ting, Su Juan, Ma Zhi-Yu, Ma Jun-Xiao, Jin Meng-Meng, Li Xiang, Lei Zhi-Hai
Nanjing Agricultural University, Nanjing 210095, China.
Nanjing Agricultural University, Nanjing 210095, China.
Gene. 2015 Jun 10;564(1):21-8. doi: 10.1016/j.gene.2015.03.038. Epub 2015 Mar 19.
Neuromedin B (NMB) is a highly conserved bombesin-related neuropeptide found in mammals. Neuromedin B (NMB) executes its effect by binding to the cell surface receptor, neuromedin B receptor (NMBR). In this study, we cloned the rabbit NMB and NMBR genes. The similarity and phylogenetic analyses of NMB and NMBR gene sequences were performed. The expression of NMB and NMBR mRNA in the rabbit was investigated using real-time RT-PCR. Our bioinformatic analysis demonstrated that the cloned rabbit NMB precursor cDNA encoded Gly-His-Phe-Met-NH2 amino acids at the C-terminus, and that its receptor possessed typical transmembrane features. The NMB mRNA was highly expressed in the CNS, while the NMBR mRNA was widely expressed in many tissues, with the highest expression in the gastrointestinal tract. The studies on the NMB distribution and function are limited by the lack of a specific antibody to this neuropeptide. In this paper, polyclonal NMB antibody was generated in mice. Western blotting analysis revealed that the prepared antibody could specifically recognize the recombinant and the endogenous NMB proteins. Immunohistochemistry analysis indicated that the NMB protein was localized in the cytoplasm of the pituitary cells. The existence of NMB protein in the hypothalamic-pituitary-gonadal axis suggests that NMB might function in rabbit reproduction.
神经介素B（NMB）是一种在哺乳动物中发现的高度保守的与蛙皮素相关的神经肽。神经介素B（NMB）通过与细胞表面受体神经介素B受体（NMBR）结合来发挥其作用。在本研究中，我们克隆了兔NMB和NMBR基因。对NMB和NMBR基因序列进行了相似性和系统发育分析。使用实时逆转录聚合酶链反应（RT-PCR）研究了兔体内NMB和NMBR mRNA的表达。我们的生物信息学分析表明，克隆的兔NMB前体cDNA在C末端编码甘氨酸-组氨酸-苯丙氨酸-甲硫氨酸-NH2氨基酸，并且其受体具有典型的跨膜特征。NMB mRNA在中枢神经系统中高表达，而NMBR mRNA在许多组织中广泛表达，在胃肠道中表达最高。由于缺乏针对这种神经肽的特异性抗体，关于NMB分布和功能的研究受到限制。在本文中，在小鼠体内产生了多克隆NMB抗体。蛋白质免疫印迹分析表明，制备的抗体可以特异性识别重组和内源性NMB蛋白。免疫组织化学分析表明，NMB蛋白定位于垂体细胞的细胞质中。下丘脑-垂体-性腺轴中存在NMB蛋白表明NMB可能在兔的生殖中发挥作用。
