Potentiation of the D2 mutant motor phenotype in mice lacking dopamine D2 and D3 receptors

doi:10.1016/S0306-4522(98)00705-2

Neuroscience

Volume 91, Issue 3, July 1999, Pages 911-924

https://doi.org/10.1016/S0306-4522(98)00705-2 Get rights and content

Abstract

Within the D₂-class of dopamine receptors, the D₂ and D₃ subtypes share the highest degree of similarity in their primary structure. However, the extent to which these two receptor subtypes have similar or different functional properties is unclear. The present study used gene targeting to generate mice deficient for D₂, D₃, and D₂/D₃ receptors. A comparative analysis of D₂ and D₃ single mutants and D₂/D₃ double mutants revealed that D₂/D₃ double mutants develop motor phenotypes that, although qualitatively similar to those seen in D₂ single mutants, are significantly more severe. Furthermore, increased levels of the dopamine metabolites dihydroxyphenyl acetic acid and homovanillic acid are found in the dorsal striatum of D₂ single mutants. The levels of these metabolites, however, are significantly higher in mice lacking D₂ and D₃ receptors. In addition, results of immunoprecipitation experiments revealed that D₂ single mutants express higher levels of D₃ receptor proteins during later stages of their postnatal development.

These results suggest that D₃ receptors compensate for some of the lacking D₂ receptor functions and that these functional properties of D₃ receptors, detected in mice with a D₂ mutant genetic background, remain masked when the abundant D₂ receptor is expressed.

Section snippets

Isolation of D₂ and D₃ receptor genes and targeted gene disruption in mouse embryonic stem cells

A 129Sv mouse genomic library was screened to isolate the genes that code for the D₂ and D₃ receptor. To isolate the D₂ gene, two oligonucleotide probes (5′-TTCGGACTCA-ACAATACAGACCAGAATGAGTGTATCATTGC-3′ (sense strand) and 5′-CAGAGTGACGATGAAGGGCACGTAGAATGAGACAATGGAGGAGTAGACCACAAAGGCAGGGTTGGTAC-3′ (anti-sense strand) corresponding to sequences that code for partially overlapping regions of the putative third cytoplasmic domain of the rat D₂ receptor³ were used for the initial screening of the

Generation of mice lacking D₂, D₃ and D₂/D₃ receptors

Gene targeting via homologous recombination was used to generate mice deficient for D₂ and D₃ receptors (see Experimental Procedures).

The targeted disruption of the D₂ gene results in a null mutation of the D₂ receptor (Fig. 1A). The male and female D₂ homozygous mutants that we generated are fertile and their offspring can be weaned three to four weeks after birth. Northern blot analysis of RNA extracted from the dorsal striatum (the anatomic region with highest density of D₂ receptor

Discussion

The present study employed gene targeting to generate D₂ and D₃ single and D₂/D₃ double mutant mice. A comparative study on these single and double mutants has identified synergistic effects of inactivation of D₂ and D₃ receptors on the impairment of locomotor activity and on the increased accumulation of dopamine metabolites in the dorsal striatum. Results from immunoprecipitation experiments further indicate an increased expression of D₃ proteins during the postnatal development of D₂ mutant

Conclusions

A comparative analysis of D₂ and D₃ single mutants and D₂/D₃ double mutants identified distinct functions of D₃ receptors that remain masked in the presence of the abundant D₂ receptor. The absence of both D₂ and D₃ receptors leads to a potentiation of some of the phenotypes that are characteristic for D₂ single mutants, namely locomotor hypoactivity and increased metabolism of striatal dopamine. Furthermore, results of immunoprecipitation studies show that D₂ mutant mice express higher levels

Acknowledgements

This work was supported by a National Science Foundation Grant IBN-9409772 (C.S.), NARSAD (C.S.), and by National Institutes of Health Grants MH51623 (C.S.) and DA08622 (J.E.P.) The early stages of this work were supported by a PHS Program Project Grant MH45212. We thank Drs Paul Good and Charles Mobbs (Mount Sinai School of Medicine) for their help and assistance with the HPLC and behavioural analyses, respectively, and Dr James Roberts (Mount Sinai School of Medicine) for his generous gift of

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^*: Both authors contributed equally to this work.

^§: Present address: Institute for Experimental Pathology, ZMBE, University of Muenster, D-48149, Germany

^**: To whom correspondence should be addressed at Columbia University, Departments of Psychiatry and Neuroscience, 1051 Riverside Drive, Box 28, New York, NY 10032, U.S.A.

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Potentiation of the D2 mutant motor phenotype in mice lacking dopamine D2 and D3 receptors

Abstract

Section snippets

Isolation of D2 and D3 receptor genes and targeted gene disruption in mouse embryonic stem cells

Generation of mice lacking D2, D3 and D2/D3 receptors

Discussion

Conclusions

Acknowledgements

Neuropsychopharmacology

Devl Brain Res.

J. biol. Chem.

Trends pharmac. Sci.

Cell

Neurosci. Lett.

Brain Res.

Cell

Cell

Cell

Neuron

A targeted mutation of the D3 dopamine receptor gene is associated with hyperactivity in mice

Proc. natn. Acad. Sci. U.S.A.

Parkinsonian-like locomotor impairment in mice lacking dopamine D2 receptors

Nature

Cloning and expression of rat D2 dopamine receptor cDNA

Nature

Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease

Biochemistry

Iodinated 2-aminotetralins and 3-amino-1-benzopyrans: ligands for dopamine D2 and D3 receptors

J. med. Chem.

Molecular diversity of the dopamine receptors

A. Rev. Pharmac. Toxicol.

Altered striatal function in a mutant mouse lacking D1A dopamine receptors

Proc. natn. Acad. Sci. U.S.A.

Potentiation of the D₂ mutant motor phenotype in mice lacking dopamine D₂ and D₃ receptors

Isolation of D₂ and D₃ receptor genes and targeted gene disruption in mouse embryonic stem cells

Generation of mice lacking D₂, D₃ and D₂/D₃ receptors

Parkinsonian-like locomotor impairment in mice lacking dopamine D₂ receptors

Cloning and expression of rat D₂ dopamine receptor cDNA

Iodinated 2-aminotetralins and 3-amino-1-benzopyrans: ligands for dopamine D₂ and D₃ receptors

Altered striatal function in a mutant mouse lacking D_1A dopamine receptors