AJP - Renal Physiology, Vol 266, Issue 4 646-F650, Copyright © 1994 by American Physiological Society

ARTICLES

Expression of the D2 subfamily of dopamine receptor genes in kidney

D. Q. Gao, L. M. Canessa, M. M. Mouradian and P. A. Jose
Georgetown University Medical Center, Washington, District of Columbia 20007.

The D2, D3, and D4 dopamine receptors cloned from brain correspond to the classically described "D2" receptors. Although radioligand binding and biochemical and functional studies have demonstrated the presence of D2-like receptors in the kidney, the expression of D2, D3, or D4 receptor genes has not been conclusively demonstrated in the kidney. Since Northern blot analysis may have precluded demonstration of dopamine receptor mRNAs because of their relative low abundance, the expression of the D2long and D3 receptor genes was studied by reverse transcription-polymerase chain reaction (RT-PCR). We were able to amplify PCR products of the predicted size using mRNA from glomeruli, proximal tubules, outer medulla, inner medulla, and renal microvessels from normotensive Wistar-Kyoto rats (WKY). Specificities of the amplified products were confirmed by restriction analysis and by sequencing the D2long product and Southern blotting the D3 product. Because some studies have suggested that D2-like receptor actions may be different between WKY and spontaneously hypertensive rats (SHR), similar studies were performed in this rat strain. In the SHR, as in WKY, PCR products of the predicted size were amplified, and restriction enzyme digestion patterns were as predicted from the cDNA sequence. The PCR-generated cDNA from the glomeruli of SHR was subcloned and sequenced and was revealed to be identical to the D2long receptor cDNA from WKY. We conclude that the D2long and D3 receptor genes are expressed in specific regions of the kidney including the glomeruli. No differences in the sequence of the D2long receptor cDNA in part of the 3rd cytoplasmic loop were noted between WKY and SHR. These studies do not rule out the possibility that mutations in other segments of the receptor exist in the SHR.

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