Naji Latifa, Carrillo-Vico Antonio, Guerrero Juan M, Calvo Juan R
Department of Medical Biochemistry and Molecular Biology, The University of Seville School of Medicine and Virgen Macarena Hospital, Avda. Sánchez Pizjuán 4, 41009 Seville, Spain.
Life Sci. 2004 Mar 19;74(18):2227-36. doi: 10.1016/j.lfs.2003.08.046.
Previous studies have shown that melatonin acts through specific receptors, including MT(1) and MT(2) membrane receptors as well as a nuclear receptor belonging to the orphan nuclear receptor family. Therefore, the goal of this study was to determine whether melatonin receptors mRNA is expressed in mouse peripheral tissues. To study the different receptors subtype expression, we have used a reverse-transcription polymerase chain reaction (RT-PCR) procedure followed by Southern hybridization with specific digoxigenin-labeled probes. RT-PCR studies revealed the presence of both MT(1) membrane receptors and ROR(alpha)1 nuclear receptors in all the peripheral tissues examined (brain, heart, lung, liver and kidney). Moreover, the expression of ROR(alpha)1 nuclear receptors was also demonstrated by Western-blot. In contrast, expression of MT(2) membrane receptors was only observed in brain and lung. These results suggest that melatonin, acting through its different subtypes receptors, plays a role in the neuroendocrine regulation of peripheral tissues function.
先前的研究表明，褪黑素通过特定受体发挥作用，包括MT(1)和MT(2)膜受体以及属于孤儿核受体家族的一种核受体。因此，本研究的目的是确定褪黑素受体mRNA是否在小鼠外周组织中表达。为了研究不同受体亚型的表达，我们采用了逆转录聚合酶链反应（RT-PCR）方法，随后用特异性地高辛标记探针进行Southern杂交。RT-PCR研究显示，在所检测的所有外周组织（脑、心脏、肺、肝脏和肾脏）中均存在MT(1)膜受体和ROR(α)1核受体。此外，通过蛋白质免疫印迹法也证实了ROR(α)1核受体的表达。相比之下，仅在脑和肺中观察到MT(2)膜受体的表达。这些结果表明，褪黑素通过其不同亚型的受体发挥作用，在外周组织功能的神经内分泌调节中起作用。
