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. 1991 Oct;11(5):437–453. doi: 10.1007/BF00734808

D2 dopamine receptors in the human retina: Cloning of cDNA and localization of mRNA

Allen Dearry 1,2,✉, Pierre Falardeau 1, Carol Shores 4, Marc G Caron 1,5,6
PMCID: PMC11567303  PMID: 1835903

Abstract

  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.

Key words: dopamine, D2 receptor, retina, cloning, in situ hybridization

References

  1. Andersen, P., Gingrich, J., Bates, M., Dearry, A., Falardeau, P., Senogles, S., and Caron, M. (1990). Dopamine receptor subtypes: Beyond the D1/D2 classification.Trends Pharmacol. Sci.11231–236. [DOI] [PubMed] [Google Scholar]
  2. Angerer, L., Cox, K., and Angerer, R. (1987). Demonstration of tissue-specific gene expression by in situ hybridization.Meth. Enzymol.152649–661. [DOI] [PubMed] [Google Scholar]
  3. Benovic, J., Bouvier, M., Caron, M., and Lefkowitz, R. (1988). Regulation of adenylyl cyclasecoupledβ-adrenergic receptors.Annu. Rev. Cell Biol.4405–428. [DOI] [PubMed] [Google Scholar]
  4. Besharse, J., Iuvone, M., and Pierce, M. (1988). Regulation of rhythmic photoreceptor metabolism: A role for post-receptoral neurons.Prog. Retin. Res.721–61. [Google Scholar]
  5. Bondy, S., Ali, S., Hong, J., Wilson, W., Fletcher, T., and Chader, G. (1983). Neurotransmitter-related features of the retinal pigment epithelium.Neurochem. Int.5285–290. [DOI] [PubMed] [Google Scholar]
  6. Brann, M., and Young, S. (1986). Dopamine receptors are located on rods in bovine retina.Neurosci. Lett.69221–226. [DOI] [PubMed] [Google Scholar]
  7. Bruinink, A., Dawis, S., Niemeyer, G., and Lichtensteiger, W. (1986). Catecholaminergic binding sites in cat retina, pigment epithelium, and choroid.Exp. Eye Res.43147–151. [DOI] [PubMed] [Google Scholar]
  8. Bunzow, J., Van Tol, H., Grandy, D., Albert, P., Salon, J., Christie, M., Machida, C., Neve, K., and Civelli, O. (1988). Cloning and expression of a rat D2 dopamine receptor cDNA.Nature336783–787. [DOI] [PubMed] [Google Scholar]
  9. Cohen, A., and Blazyniski, C. (1990). Dopamine and its agonists reduce a light-sensitive pool of cyclic AMP in mouse photoreceptors.Vis. Neurosci.443–52. [DOI] [PubMed] [Google Scholar]
  10. Cullen, B. (1987). Use of eukaryotic expression technology in the functional analysis of cloned genes.Meth. Enzymol.152684–704. [DOI] [PubMed] [Google Scholar]
  11. Dal Toso, R., Sommer, B., Evert, M., Herb, A., Pritchett, D., Bach, A., Shivers, B., and Seeburg, P. (1989). The dopamine D2 receptor: Two molecular forms generated by alternative splicing.EMBO J.84025–4034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dawis, S., and Niemeyer, G. (1986). Dopamine influences the light peak of the perfused mammalian eye.Invest. Ophthalmol. Vis. Sci.27330–335. [PubMed] [Google Scholar]
  13. Dearry, A., and Burnside, B. (1985). Dopamine inhibits forskolin- and 3-isobutyl-1-methylxanthineinduced dark-adaptive retinomotor movements in isolated telost retinas.J. Neurochem.441753–1763. [DOI] [PubMed] [Google Scholar]
  14. Dearry, A., and Burnside, B. (1986). Dopaminergic regulation of cone retinomotor movement in isolated teleost retinas. I. Induction of cone contraction is mediated by D2 receptors.J. Neurochem.461006–1021. [DOI] [PubMed] [Google Scholar]
  15. Dearry, A., and Burnside, B. (1988). Dopamine induces light-adaptive retinomotor movements in teleost photoreceptors and retinal pigment epithelium. InDopaminergic Mechanisms in Vision (I. Bodis-Wollner Ed), Alan R. Liss, New York, pp. 109–135. [Google Scholar]
  16. Dearry, A., and Burnside, B. (1989). Light-induced dopamine release from teleost retinas acts as a light-adaptive signal to the retinal pigment epithelium.J. Neurochem.53870–878. [DOI] [PubMed] [Google Scholar]
  17. Dearry, A., Edelman, J., Miller, S., and Burnside, B. (1990a). Dopamine induces light-adaptive retinomotor movements in bullfrog cones via D2 receptors and in retinal pigment epithelium via D1 receptors.J. Neurochem.541367–1378. [DOI] [PubMed] [Google Scholar]
  18. Dearry, A., Gingrich, J., Falardeau, P., Fremeau, R., Bates, M., and Caron, M. (1990b). Molecular cloning and expression of the gene for a human D1 dopamine receptor.Nature34772–76. [DOI] [PubMed] [Google Scholar]
  19. Delean, A., Hancock, A., and Lefkowitz, R. (1982). Validation and statistical analysis of a computer modeling method for quantitative analysis of radioligand binding data for mixtures of pharmacological receptor subtypes.Mol. Pharmacol.215–16. [PubMed] [Google Scholar]
  20. Dickman, M., and Tran, V. (1990). D1 and D2 dopamine receptors are differentially localized in the rat retina.Invest. Ophthalmol. Vis. Sci. Suppl.31331. [Google Scholar]
  21. Dowling, J. (1986). Dopamine: A retinal neuromodulator?Trends Neurosci.9236–240. [Google Scholar]
  22. Dubocovich, M. (1984). Modulation of [3H]dopamine release from rabbit retina.Fed. Proc.432714–2718. [PubMed] [Google Scholar]
  23. Eder, D., Gilbreath, M., and Williams, T. (1990). Reduction of late PIII responses by dopamine in isolated rat retina.Invest. Ophthalmol. Vis. Sci. Suppl.31388. [Google Scholar]
  24. Ehinger, B. (1983). Functional role of dopamine in the retina.Prog. Retinal Res.2213–232. [Google Scholar]
  25. Frederick, J., Rayborn, M., Laties, A., Lam, D., and Hollyfield, J. (1982). Dopaminergic neurons in the human retina.J. Comp. Neurol.21065–79. [DOI] [PubMed] [Google Scholar]
  26. Fremeau, R., Lundblad, J., Pritchett, D., Wilcox, J., and Roberts, J. (1986). Regulation of pro-opiomelanocortin gene transcription in individual cell nuclei.Science2341265–1269. [DOI] [PubMed] [Google Scholar]
  27. Giros, B., Sokoloff, P., Martres, M., Riou, J., Emorine, L., and Schwartz, J. (1989). Alternative splicing directs the expression of two D2 dopamine receptor isoforms.Nature342923–926. [DOI] [PubMed] [Google Scholar]
  28. Grandy, D., Marchionni, M., Makam, H., Stofko, R., Alfano, M., Frothingham, L., Fischer, J., Burke-Howie, K., Bunzow, J., Server, A., and Civelli, O. (1989). Cloning of the cDNA and gene for a human D2 dopamine receptor.Proc. Natl. Acad. Sci. USA869762–9766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Grant, G., and Werblin, F. (1990). Dopaminergic modulation of a sustained, voltage-gated calcium current in vertebrate rods of the tiger salamander.Invest. Ophthalmol. Vis. Sci. Suppl.31176. [Google Scholar]
  30. Gredal, O., Parkinson, D., and Nielsen, M. (1987). Binding of [3H]SCH23390 to dopamine D1 receptors in rat retina in vitro.Eur. J. Pharmacol.137241–245. [DOI] [PubMed] [Google Scholar]
  31. Grigoriadis, D., Niznik, H., Jarvie, K., and Seeman, P. (1988). Glycoprotein nature of D2 dopamine receptors.FEBS Lett.227220–224. [DOI] [PubMed] [Google Scholar]
  32. Hensler, J., Cotterell, D., and Dubocovich, M. (1987). Pharmacological and biochemical characterization of the D1 dopamine receptor mediating acetylcholine release in rabbit retinas.J. Pharmacol. Exp. Ther.243857–867. [PubMed] [Google Scholar]
  33. Huff, R., and Molinoff, P. (1982). Quantitative determination of dopamine receptor subtypes not linked to activation of adenylate cyclase in rat striatum.Proc. Natl. Acad. Sci. USA797561–7565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Iuvone, M., Boatright, J., and Bloom, M. (1987). Dopamine mediates the light-evoked suppression of serotonin N-acetyltransferase activity in retina.Brain Res.418314–324. [DOI] [PubMed] [Google Scholar]
  35. Jensen, R., and Daw, N. (1986). Effects of dopamine and its agonists and antagonsits on the receptive field properties of ganglion cells in the rabbit retina.Neuroscience17837–855. [DOI] [PubMed] [Google Scholar]
  36. Kebabian, J., and Calne, D. (1979). Multiple receptors for dopamine.Nature27793–96. [DOI] [PubMed] [Google Scholar]
  37. Kimelman, D., and Kirschner, M. (1989). An antisense mRNA directs the covalent modification of the transcript encoding fibroblast growth factor in Xenopus oocytes.Cell59687–696. [DOI] [PubMed] [Google Scholar]
  38. Land, H., Grez, H., Hauser, H., Lindenmaier, W., and Schutz, G. (1981). 5′-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency.Nucleic Acids Res.92251–2266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lasater, E., and Dowling, J. (1985). Dopamine decreases conductance of the electrical junctions between cultured retinal horizontal cells.Proc. Natl. Acad. Sci. USA823025–3029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lefkowitz, R., and Caron, M. (1988). The adrenergic receptors: Models for the study of receptors coupled to guanine nucleotide regulatory proteins.J. Biol. Chem.2634993–4996. [PubMed] [Google Scholar]
  41. Lin, C., McGonigle, P., and Molinoff, P. (1987). Characterization of D2 dopamine receptors in a tumor of the rat anterior pituitary gland.J. Pharmacol. Exp. Ther.242950–956. [PubMed] [Google Scholar]
  42. McGonigle, P., Wax, M., and Molinoff, P. (1988). Characterization of binding sites for [3H]spiroperidol in human retina.Invest. Ophthalmol. Vis. Sci.29687–694. [PubMed] [Google Scholar]
  43. Monsma, F., McVittie, L., Gerfen, C., Mahan, L., and Sibley, D. (1989). Multiple D2 dopamine receptors produced by alternative RNA splicing.Nature342926–929. [DOI] [PubMed] [Google Scholar]
  44. O'Dowd, B., Hnatowich, M., Caron, M., Lefkowitz, R., and Bouvier, M. (1989). Palmitoylation of the humanβ2-adrenergic receptor.J. Biol. Chem.2647564–7569. [PubMed] [Google Scholar]
  45. Ovchinnikov, Y., Abdulaev, N., and Bogachuk, A. (1988). Two adjacent cysteine residues in the C-terminal cytoplasmic fragment of bovine rhodopsin are palmitoylated.FEBS Lett.2301–5. [DOI] [PubMed] [Google Scholar]
  46. Piccolino, M., Demontis, G., Witkovsky, P., Strettoi, E., Cappagli, G., Porceddu, M., DeMontis, M., Pepitoni, S., Biggio, G., Meller, E., and Bohmaker, K. (1989). Involvement of D1 and D2 dopamine receptors in the control of horizontal cell electrical coupling in the turtle retina.Eur. J. Neurosci.1247–257. [DOI] [PubMed] [Google Scholar]
  47. Pierce, M., and Besharse, J. (1985). Circadian regulation of retinomotor movements. I. Interaction of melatonin and dopamine in the control of cone length.J. Gen. Physiol.86671–689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Qu, Z., Fertel, R., Neff, N., and Hadjiconstantinou, M. (1989). Pharmacological characterization of rat retinal dopamine receptors.J. Pharmacol. Exp. Ther.248621–625. [PubMed] [Google Scholar]
  49. Regan, J., Kobilka, T., Yang-Feng, T., Caron, M., Lefkowitz, R., and Kobilka, B. (1988). Cloning and expression of a human kidney cDNA for anα2-adrenergic receptor subtype.Proc. Natl. Acad. Sci. USA856301–6305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Reme, C., Wirz-Justice, A., Rhyner, A., and Hofman, S. (1986). Circadian rhythm in the light response of rat retinal disk-shedding and autophagy.Brain Res.369356–360. [DOI] [PubMed] [Google Scholar]
  51. Robbins, J., Wakakuwa, K., and Ikeda, H. (1988). Noradrenaline action on cat retinal ganglion cells is mediated by dopamine (D2) receptors.Brain Res.43852–60. [DOI] [PubMed] [Google Scholar]
  52. Rossetti, Z., Silvia, C., Krajac, D., Neff, N., and Hadjiconstantinou, M. (1990). Aromatic L-amino acid decarboxylase is modulated by D1 dopamine receptors in rat retina.J. Neurochem.54787–791. [DOI] [PubMed] [Google Scholar]
  53. Sambrook, J., Fritsch, E., and Maniatis, T. (1989).Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. [Google Scholar]
  54. Sato, T., Yoneyama, T., Kim, H., and Suzuki, T. (1987). Effect of dopamine and haloperidol on the c-wave and light peak of light-induced retinal responses in chick eye.Doc. Ophthalmol.6587–95. [DOI] [PubMed] [Google Scholar]
  55. Seeman, P. (1981). Brain dopamine receptors.Pharmacol. Rev.32229–313. [PubMed] [Google Scholar]
  56. Stormann, T., Gdula, D., Weiner, D., and Brann, M. (1990). Molecular cloning and expression of a dopamine D2 receptor from human retina.Mol. Pharmacol.371–6. [PubMed] [Google Scholar]
  57. Teranishi, T., Negishi, K., and Kato, S. (1984). Regulatory effect of dopamine on spatial properties of horizontal cells in carp retina.J. Neurosci.41271–1280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Todd, R., Khurana, T., Sajovic, P., Stone, K., and O'Malley, K. (1989). Cloning of ligand-specific cell lines via gene transfer: Identification of a D2 dopamine receptor subtype.Proc. Natl. Acad. Sci. USA8610134–10138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Vaney, D. (1985). The morphology and topographic distribution of AII amacrine cells in the cat retina.Proc. Roy. Soc. Lond. B224475–488. [DOI] [PubMed] [Google Scholar]
  60. Witkovsky, P., Stone, S., and Tranchina, D. (1989). Photoreceptor to horizontal cell synaptic transfer in the Xenopus retina: Modulation by dopamine ligands and a circuit model for interactions of rod and cone inputs.J. Neurophysiol.62864–881. [DOI] [PubMed] [Google Scholar]
  61. Zarbin, M., Wamsley, J., Palacios, J., and Kuhar, M. (1986). Autoradiographic localization of high affinity GABA, benzodiazepine, dopaminergic, adrenergic, and muscarinic cholinergic receptors in the rat, monkey, and human retina.Brain Res.37475–92. [DOI] [PubMed] [Google Scholar]
  62. Zawilska, J., and Iuvone, M. (1989). Catecholamine receptors regulating serotonin N-acetyltransferase activity and melatonin content of chicken retina and pineal gland: D2-dopamine receptors in retina andα2-adrenergic receptors in pineal gland.J. Pharmacol. Exp. Ther.25086–92. [PubMed] [Google Scholar]

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