title: Cloning of neuromedin B and its receptor in the rabbit and generating a polyclonal antibody to the neuromedin B protein.
abstract: 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.
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Ting-Ting
authors: TT
authors: Guo
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Juan
authors: J
authors: Su
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Zhi-Yu
authors: ZY
authors: Ma
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Jun-Xiao
authors: JX
authors: Ma
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Meng-Meng
authors: MM
authors: Jin
authors: Nanjing Agricultural University, Nanjing 210095, China.
authors: Xiang
authors: X
authors: Li
authors: Nanjing Agricultural University, Nanjing 210095, China. Electronic address: lei55062@163.com.
authors: Zhi-Hai
authors: ZH
authors: Lei
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood*; D002490:Central Nervous System / Q000378:metabolism; D003001:Cloning, Molecular; D005260:Female; D015870:Gene Expression; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D015287:Neurokinin B / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D009928:Organ Specificity; D011817:Rabbits; D018004:Receptors, Bombesin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA
chemical_list: D000906:Antibodies; D018004:Receptors, Bombesin; D015287:Neurokinin B; C038393:neuromedin B
keywords: Distribution; NMB; NMBR; Polyclonal antibody; Rabbit; cDNA cloning