Characterization of NMB, GRP and their receptors (BRS3, NMBR and GRPR) in chickens
Characterization of NMB, GRP and their receptors (BRS3, NMBR and GRPR) in chickens
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鸡 NMB、GRP 及其受体（BRS3、NMBR 和 GRPR）的表征
DOI：
10.1530/jme-17-0020
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发表时间：
2017-07-01
影响因子：
3.5
通讯作者：
Wang, Yajun
中科院分区：
文献类型：
--
作者：
Mo, Chunheng;Huang, Long;Wang, Yajun
胃泌素释放肽（GRP）和神经介肽B（NMB）是两种结构和功能相关的多肽，在哺乳动物的许多生理和病理过程中起着重要作用。然而，关于禽类NMB、GRP及其受体的表达和功能的信息是有限的。在这里，我们在鸡中表征cNMB、cGRP及其受体（cNMBR、cGRPR和cBRS 3）。我们的研究结果表明：（1）CHO细胞表达的cNMBR和cGRPR分别被cNMB和cGRP激活，表明cNMBR和cGRPR是cNMB和cGRP特异性受体;惊人的是，BRS 3的鸡（/spotted加尔斯），其与小鼠铃蟾肽受体亚型3（BRS 3）直接相关，可被GRP和NMB有效激活，证明这两种肽都是鸡的内源性配体，（/spotted gar）BRS 3;（2）实时荧光定量PCR（qPCR）结果显示，cGRPR在鸡的卵巢、胰腺、腺胃、脊髓和脑组织中表达丰富，而cNMB，cNMBR和cBRS 3主要在脑和睾丸中表达;（3）qPCR、Western blot和免疫组化结果显示cGRP主要在垂体前叶表达，且主要定位于LH细胞，提示cGRP可能是一种新的鸡垂体激素。总之，我们的数据有助于揭示GRP，NMB及其受体在鸟类中的作用，并从进化的角度提供了第一个有说服力的证据，即在脊椎动物中，GRP和NMB是BRS 3的内源性配体，BRS 3是一种孤儿受体，困扰了内分泌学家二十多年。
The two structurally and functionally related peptides, gastrin-releasing peptide (GRP) and neuromedin B (NMB) play critical roles in many physiological/pathological processes in mammals. However, the information regarding the expression and functionality of avian NMB, GRP and their receptors is limited. Here, we characterized cNMB, cGRP and their receptors (cNMBR, cGRPR and cBRS3) in chickens. Our results showed that: (1) cNMBR and cGRPR expressed in CHO cells could be potently activated by cNMB and cGRP, respectively, as monitored by cell-based luciferase reporter assays, indicating that cNMBR and cGRPR are cNMB- and cGRP-specific receptors; strikingly, BRS3 of chickens (/spotted gars), which is orthologous to mouse bombesin receptor subtype-3 (BRS3), could be potently activated by GRP and NMB, demonstrating that both peptides are the endogenous ligands for chicken (/spotted gar) BRS3; (2) quantitative real-time PCR (qPCR) revealed that cGRPR is widely expressed in chicken tissues with abundant expression in the ovary, pancreas, proventriculus, spinal cord and brain, whereas cNMB, cNMBR and cBRS3 are mainly expressed in the brain and testes; (3) interestingly, qPCR, Western blot and immunostaining revealed that cGRP is predominantly expressed in the anterior pituitary and mainly localized to LH-cells, suggesting that cGRP is likely a novel pituitary hormone in chickens. In summary, our data help to uncover the roles of GRP, NMB and their receptors in birds, and provide the first persuasive evidence from an evolutionary prospective that in vertebrates, GRP and NMB are the endogenous ligands for BRS3, an orphan receptor that has puzzled endocrinologists for more than two decades.