D2 dopamine receptors in the human retina: cloning of cDNA and localization of mRNA.
1. We have obtained a cDNA clone encoding a human retinal D2 dopamine receptor. 2. The longest open reading frame (1242 bp) of this clone encodes a protein of 414 amino acids having a predicted molecular weight of 47,000 and a transmembrane topology similar to that of other G protein-coupled receptors. 3. Transient transfection of COS-7 cells with an expression vector containing the clone resulted in expression of a protein possessing a pharmacological profile similar to that of the D2 dopamine receptor found in striatum and retina. 4. Northern blot analysis indicated that, in rat brain and retina, the mRNA for this receptor was 2.9 kb in size. 5. In situ hybridization was performed to examine the distribution of the mRNA for this receptor in human retina. Specific hybridization was detected in both the inner and the outer nuclear layers. 6. These findings are consistent with prior physiological and autoradiographic studies describing the localization of D2 dopamine receptors in vertebrate retinas. Our observations suggest that photoreceptors as well as cells in the inner nuclear layer of human retinas may express the mRNA for this D2 dopamine receptor.
Dearry
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Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710.
Falardeau
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Shores
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Caron
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004298:Dopamine / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D008961:Models, Structural; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D011954:Receptors, Dopamine / Q000235:genetics* / Q000378:metabolism; D017448:Receptors, Dopamine D2; D011994:Recombinant Proteins / Q000378:metabolism; D012160:Retina / Q000166:cytology / Q000502:physiology*; D013134:Spiperone / Q000378:metabolism; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2; D011994:Recombinant Proteins; D013134:Spiperone; D004247:DNA; D004298:Dopamine