Yamaguchi I, Jose P A, Mouradian M M, Canessa L M, Monsma F J, Sibley D R, Takeyasu K, Felder R A
Department of Pathology, University of Virginia Medical Center, Charlottesville 22908.
Am J Physiol. 1993 Feb;264(2 Pt 2):F280-5. doi: 10.1152/ajprenal.1993.264.2.F280.
The existence of dopamine receptor subtypes outside the central nervous system has been suggested by pharmacological and biochemical techniques. Whether the renal dopamine receptors are distinct from those cloned from the brain remains to be determined. Indeed, the expression of any of these receptor genes in the kidney has not been demonstrated definitively. In this study, we amplified D1A receptor cDNA from microdissected proximal convoluted tubules of the rat kidney by reverse transcription-polymerase chain reaction; primers were based on the sequence of rat D1A cDNA cloned from the brain and corresponded to the third cytoplasmic loop of the receptor. Specificity of the amplified products was verified by restriction analysis, Southern blots, and sequencing. Furthermore, solution hybridization indicated the presence of a single ribonuclease-protected RNA species corresponding to the D1A receptor mRNA in proximal tubules. These studies report for the first time the presence of D1A receptor message in the kidney.
药理学和生化技术表明中枢神经系统外存在多巴胺受体亚型。肾多巴胺受体是否与从大脑克隆的受体不同仍有待确定。事实上,这些受体基因在肾脏中的任何一种表达尚未得到确切证实。在本研究中,我们通过逆转录-聚合酶链反应从大鼠肾脏显微切割的近端曲管中扩增了D1A受体cDNA;引物基于从大脑克隆的大鼠D1A cDNA序列,对应于受体的第三个细胞质环。通过限制性分析、Southern印迹和测序验证了扩增产物的特异性。此外,溶液杂交表明近端小管中存在一种与D1A受体mRNA相对应的单核糖核酸酶保护RNA物种。这些研究首次报道了肾脏中存在D1A受体信息。