Cloning and sequence analysis of brain cDNA encoding a Xenopus D2 dopamine receptor

Martens, Gerard J.M.; Molhuizen, H.O.F.; Gröneveld, D.; Roubos, Eric W.

doi:10.1016/0014-5793(91)80364-9

Cited by 30 publications

(20 citation statements)

References 31 publications

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“…An array of experimental approaches [Verburg-Van Kemenade et al, 1986;Martens et al, 1991;Morra et al, 1992;Sugamori et al, 1994;Tonosaki et al, 1995;Duchamp-Viret et al, 1997;Alonso-Gomez et al, 2000] suggest that amphibians, like mammals, have D1-and D2-like dopamine receptors. Unfortunately, the functional properties of amphibian forebrain dopamine receptors remain largely uninvestigated.…”

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confidence: 99%

Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, Rana pipiens

Chu

Wilczynski

Wilcox³

2001

Brain Behav Evol

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The pharmacological profiles of D1- and D2-like dopamine receptors were investigated for native brain receptors in the leopard frog, Rana pipiens, using direct binding assays, which characterize functional receptors rather than assess total receptor protein. We used homogenate assays of R. pipiens fore- and midbrains to determine, via saturation isotherms, that the dissociation constant, Kd, for ³H-SCH-23390 binding to the D1-like receptors was 0.29 nM, and the maximal receptor density, Bmax, was 40 fmoles/mg protein. This compares with the more than 10-fold higher density of D1 sites in rat striatum. Specific binding for the D2-like receptors was measurable using these methods with ³H-spiperone as the ligand. However, saturation of binding was not achieved. This contrasts with the > 400 fmoles/mg protein Bmax in rat striatum. Pharmacological profiles (rank order of potency of displacing drugs) for each receptor type were determined. We used non-radioactive SCH-23390, SKF-38393, sulpiride, and spiperone to displace ³H-SCH-23390 and ³H-spiperone at D1 and D2 receptors, respectively. Parallel displacement assays were performed with rat striatal controls. Results indicated that the relative rank order displacements in anuran dopamine receptors were characteristic of D1- and D2-like receptors. However, the rank orders were not identical to those in mammals. The rank order for affinity at D1-like receptors in both rats and frogs was SCH-23390 > SKF-38393 > spiperone > sulpiride. The rank order for affinity at D2-like receptors was spiperone > SCH-23390 > sulpiride > SKF-38393 in frogs, and spiperone > sulpiride > SCH-23390 > SKF-38393 in rats. SKF-38393 and spiperone had similar affinities for the ‘D1’ receptors in both species. SCH-23390 had a slightly lower affinity for the D1-like receptors in Rana, whereas sulpiride had a significantly lower affinity for Rana D1-like receptors compared to rat D1 receptors. In Rana D2-like receptors, spiperone and sulpiride were significantly less potent compared to rat. However, SCH-23390 and SKF-38393 were equally potent for the D2-like receptors in both species. The results indicate that amphibian brain dopamine receptors fall into two classes similar to the mammalian D1 and D2 subfamilies, but with binding characteristics slightly different from those typically described in mammals. This work represents the first pharmacological characterization of native brain dopaminergic receptors in an anuran amphibian. Because direct binding assays measure the initial aspect of the functional interaction between transmitter and receptor, these data provide an important complement to studies using cell expression systems.

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Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, Rana pipiens

Chu

Wilczynski

Wilcox³

2001

Brain Behav Evol

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“…Upon subcloning and sequencing, a cDNA clone (clone pE4RlXD2) structurally different from the previously reported Xenopus brain D, dopamine receptor cDNA clone (clone pHRlXD2) [17] was found. Fig.…”

Section: Resultsmentioning

confidence: 71%

“…A Xenopus brain cDNA library in the vector Agtll (generously provided by Drs K. Richter and G. Kreil, Salzburg, Austria) was screened as previously described [17]. The hybridization probe was a nearly full-length rat brain cDNA encoding the small isoform of the D, dopamine receptor (clone pGRB-2, kindly provided by Dr. 0.…”

Section: Screening Of Xenopus Brain Cdna Librarymentioning

confidence: 99%

“…The question arises whether the alternative-splicing event generating D, receptor diversity is restricted to mammals or if it also occurs in lower vertebrates. We recently reported the cloning of a brain cDNA encoding the large isoform of the D, dopamine receptor of the South-African clawed toad Xenopus laevis [17]. Accordingly, we were interested in examining the possible occurrence of another form of the D, dopamine receptor in Xenopus.…”

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Expression of the Xenopus D₂ dopamine receptor

Martens

Groenen

Gröneveld

et al. 1993

European Journal of Biochemistry

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In the amphibian Xenopus Zuevis the D, dopamine receptor is involved in the regulation of the melanotrope cells of the intermediate pituitary during background adaptation of the animal. The Xenopus D, receptor has been found to be pharmacologically different from the mammalian D, receptor. In a number of mammalian species alternative splicing generates two molecular forms of the D, receptor. These isoforms differ by the presence or absence of 29 amino acids in the third cytoplasmic loop which is thought to be involved in guanine-nucleotide-binding-regulatory-protein (G-protein) binding of the receptor. We previously described a cDNA encoding the large isoform of the Xenopus D, receptor. Here we report on the isolation of a brain cDNA encoding a second, structurally different Xenopus D, dopamine receptor. Both Xenopus receptors correspond to the large isoform of the D, receptor and they display a high degree of sequence identity with their mammalian counterparts. Their occurrence reflects the expression of two Xenopus D, receptor genes and they are expressed to approximately the same level. In contrast to mammals, PCR analysis gave no evidence for alternative splicing during D, receptor expression in Xenopus brain and pituitary. Tissue-specific expression of the Xenopus D, receptor was observed in the pituitary during background adaptation. The low level of receptor mRNA in melanotrope cells of white animals compared to that of black animals may be caused by chronic dopamine stimulation of melanotrope cells in white animals with consequent cellular desensitization and down regulation of the D, receptor gene.Recent cloning of cDNA and genomic DNA fragments has revealed the existence of a number of dopamine-receptor subtypes, namely the D1, D,, DS, D, and D, receptors [l-71. These receptors, which belong to the family of guaninenucleotide-binding-regulatory-protein(G-protein)-coupled receptors, are characterized by the presence of seven putative membrane-spanning domains within their structures [S]. The diversity of dopamine receptors is probably generated by gene duplication events. The D, and D, receptors are structurally and functionally related, and they are encoded by intronless genes. In contrast, the genes encoding the structurally and functionally related D,, D, and D, receptors all contain a number of introns. The structural and functional relationships between the dopamine-receptor subtypes D, and D, on the one hand, and DZ, D, and D4 on the other, may reflect their evolutionary history. In human, bovine, rat and Note. The novel nucleotide sequence data published here has beeen deposited with the EMBL sequence data banks and is available under accession number X72902. mouse the degree of diversity of dopamine-receptor subtypes is further increased by alternative splicing of D,-dopaminereceptor-gene transcripts which results in D, , and D,, receptor subtypes [9-131. The D, receptor isoforms differ from each other by the presence or absence of 29 amino-acid residues in the third cytoplasmic loop. Cells tran...

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“…preproinsulins, proopiomelanocortins, and proenkephalins which share 92-95% nucleotide and amino acid identity (42)(43)(44)(45)(46). The low homologies displayed between the members of the Xenopus D1 receptor family suggest that they diverged prior to the predicted duplication event, in accord with the proposed early evolutionary appearance (=500 million years) and ancient status of the DI-like receptor subfamily (33).…”

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confidence: 49%

D1A, D1B, and D1C dopamine receptors from Xenopus laevis.

Sugamori

Demchyshyn

Chung

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Cloning and sequence analysis of brain cDNA encoding a Xenopus D₂ dopamine receptor

Abstract: To determine the hybridization conditions for library screening,Published by Elsevier Science Publishers B. V.

Cited by 30 publications

References 31 publications

Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, <i>Rana pipiens</i>

Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, <i>Rana pipiens</i>

Expression of the Xenopus D₂ dopamine receptor

D1A, D1B, and D1C dopamine receptors from Xenopus laevis.

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Cloning and sequence analysis of brain cDNA encoding a Xenopus D2 dopamine receptor

Abstract: To determine the hybridization conditions for library screening,Published by Elsevier Science Publishers B. V.

Cited by 30 publications

References 31 publications

Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, <i>Rana pipiens</i>

Pharmacological Characterization of the D1- and D2-Like Dopamine Receptors from the Brain of the Leopard Frog, <i>Rana pipiens</i>

Expression of the Xenopus D2 dopamine receptor

D1A, D1B, and D1C dopamine receptors from Xenopus laevis.

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Product

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Cloning and sequence analysis of brain cDNA encoding a Xenopus D₂ dopamine receptor

Expression of the Xenopus D₂ dopamine receptor