CLONING AND PHARMACOLOGICAL CHARACTERIZATION OF A RAT KAPPA-OPIOID RECEPTOR

被引:359
作者
MENG, F [1 ]
XIE, GX [1 ]
THOMPSON, RC [1 ]
MANSOUR, A [1 ]
GOLDSTEIN, A [1 ]
WATSON, SJ [1 ]
AKIL, H [1 ]
机构
[1] STANFORD UNIV, STANFORD, CA 94305 USA
关键词
GUANINE NUCLEOTIDE-BINDING PROTEIN-COUPLED RECEPTOR; DYNORPHIN; CAMP; IN-SITU HYBRIDIZATION;
D O I
10.1073/pnas.90.21.9954
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A full-length cDNA was isolated from a rat striatal library by using low-stringency screening with two PCR fragments, one spanning transmembrane domains 3-6 of the mouse delta opioid receptor and the other unidentified but homologous to the mouse delta receptor from rat brain. The novel cDNA had a long open reading frame encoding a protein of 380 residues with 59% identity to the mouse delta receptor and topography consistent with a seven-helix guanine nucleotide-binding protein-coupled receptor. COS-1 cells transfected with the coding region of this clone showed high-affinity binding to kappa opioid receptor-selective ligands such as dynorphin A and U-50,488 and also nonselective opioid ligands such as bremazocine, ethylketocyclazocine, and naloxone. Not bound at all (or bound with low affinity) were dynorphin A-(2-13), enantiomers of naloxone and levophanol [i.e., (+)-naloxone and dextrorphan], and selective mu and delta opioid receptor ligands. Activation of the expressed receptor by kappa receptor agonists led to inhibition of cAMP. Finally, in situ hybridization revealed a mRNA distribution in rat brain that corresponded well to the distribution of binding sites labeled with kappa-selective ligands. These observations indicate that we have cloned a cDNA encoding a rat kappa receptor of the kappa1 subtype.
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页码:9954 / 9958
页数:5
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