Nakagawa Atsushi, Satake Hanae, Nakabayashi Hajime, Nishizawa Makoto, Furuya Keisuke, Nakano Shigeru, Kigoshi Toshikazu, Nakayama Kohzo, Uchida Kenzo
Department of Internal Medicine, Kanazawa Medical University, Daigaku 1-1, Uchinada 920-0293, Japan.
Auton Neurosci. 2004 Jan 30;110(1):36-43. doi: 10.1016/j.autneu.2003.11.001.
We previously reported that afferent signals of the rat hepatic vagus increased upon intraportal appearance of insulinotropic hormone glucagon-like peptide-1(7-36) amide (GLP-1), but not glucose-dependent insulinotropic polypeptide (GIP). To obtain molecular evidence for the vagal chemoreception of GLP-1, the concept derived from those electrophysiological observations, receptor gene expressions of GLP-1 and GIP in the rat nodose ganglion were examined by means of reverse transcriptase-mediated polymerase chain reaction (RT-PCR) and Northern blot analysis. Gene expression of the GLP-1 receptor was clearly detected by both RT-PCR and Northern blot analysis. In situ hybridization study confirmed that the expression occurs in neuronal cells of the ganglion. As to the GIP receptor, RT-PCR amplified the gene transcript faintly though Northern blot analysis failed to detect any messages. However, semi-quantitative RT-PCR revealed that the ratio of the gene expression level of the GIP receptor to that of the GLP-1 receptor was less than 1:250, indicating that receptor gene expression of GIP is practically negligible in the ganglion. Additionally, an equal level of GLP-1 receptor gene expressions between left- and right-side ganglia was evidenced by semi-quantitative RT-PCR, implying possible extrahepatic occurrence of vagal GLP-1 reception in addition to the reception through the hepatic vagus (originating from the left-side ganglion). The present results offer, for the first time, the molecular basis for the vagal chemoreception of GLP-1 via its specific receptor.
我们之前报道过,当促胰岛素激素胰高血糖素样肽-1(7-36)酰胺(GLP-1)经门静脉注入时,大鼠肝迷走神经的传入信号增强,而葡萄糖依赖性促胰岛素多肽(GIP)注入时则不然。为了获得GLP-1迷走化学感受的分子证据,基于那些电生理观察结果提出这一概念,我们通过逆转录介导的聚合酶链反应(RT-PCR)和Northern印迹分析,检测了大鼠结状神经节中GLP-1和GIP的受体基因表达。通过RT-PCR和Northern印迹分析均清楚地检测到了GLP-1受体的基因表达。原位杂交研究证实该表达发生在神经节的神经细胞中。至于GIP受体,RT-PCR微弱地扩增出了基因转录本,不过Northern印迹分析未能检测到任何信息。然而,半定量RT-PCR显示,GIP受体基因表达水平与GLP-1受体基因表达水平之比小于1:250,这表明神经节中GIP的受体基因表达实际上可忽略不计。此外,半定量RT-PCR证明左右侧神经节之间GLP-1受体基因表达水平相当,这意味着除了通过肝迷走神经(起源于左侧神经节)进行感受外,迷走神经对GLP-1的感受可能在肝外发生。本研究结果首次为GLP-1通过其特异性受体进行迷走化学感受提供了分子基础。