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Dopamine D4 receptor isoform mRNA and protein are expressed in the rat cortical collecting duct.

机译:多巴胺D4受体同工型mRNA和蛋白在大鼠皮质收集管中表达。

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We reported previously [Am. J. Physiol. 271 (Renal Fluid Electrolyte Physiol. 40): F391-F400, 1996] that dopamine inhibits vasopressin (AVP)-dependent water permeability and Na+ transport in the rat cortical collecting duct (CCD) apparently through a D4 dopamine receptor. The present experiments used RT-PCR of total RNA extracted from microdissected rat CCD to determine whether the D4 and D1A dopamine receptor isoforms are expressed. Specific primers were used to amplify three regions of the D4 cDNA. All three gave products with 98-100% nucleotide identity to the known rat D4 sequence; however, there was an extra 6-bp insert at the 3' end of the second transmembrane region that was identical to the human and mouse sequences but which had not been documented in the rat sequence. D4 receptor protein was also localized exclusively to the CCD and medullary collecting ducts by immunohistochemistry. Two regions of the D1A dopamine receptor message were also amplified by RT-PCR of RNA from rat CCD and were verified by sequencing and immunohistochemistry. We conclude that both D4 and D1A dopamine receptors are expressed in the rat CCD, but the physiological effects are attributable to a D4 receptor.
机译:我们以前曾报道过[Am。 J.生理学。 271(Renal Fluid Electrotete Physiol。40:F391-F400,1996),即多巴胺显然通过D4多巴胺受体抑制了大鼠加压素(AVP)依赖的水渗透性和Na +在大鼠皮质收集管(CCD)中的运输。本实验对从显微解剖的大鼠CCD提取的总RNA进行了RT-PCR,以确定D4和D1A多巴胺受体同工型是否表达。特异性引物用于扩增D4 cDNA的三个区域。所有这三个产物均与已知的大鼠D4序列具有98-100%的核苷酸同一性。然而,在第二个跨膜区的3'末端有一个额外的6 bp插入片段,与人和小鼠序列相同,但在大鼠序列中没有记载。 D4受体蛋白还通过免疫组织化学专门定位于CCD和髓样收集管。 D1A多巴胺受体信息的两个区域也通过来自大鼠CCD的RNA的RT-PCR进行了扩增,并通过测序和免疫组化进行了验证。我们得出的结论是D4和D1A多巴胺受体均在大鼠CCD中表达,但生理作用可归因于D4受体。

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