Cloning and expression of a rat D2 dopamine receptor cDNA

Abstract

Dopamine receptors are classified into D1 and D2 subtypes on the basis of their pharmacological and biochemical characteristics. The D2 dopamine receptor has been implicated in the pathophysiology and treatment of movement disorders, schizophrenia and drug addiction. The D2 dopamine receptor interacts with guanine nucleotide-binding proteins to induce second messenger systems. Other members of the family of receptors that are coupled to G proteins share a significant similarity in primary amino-acid sequence and exhibit an archetypical topology predicted to consist of seven putative transmembrane domains. We have taken advantage of the expected nucleotide sequence similarities among members of this gene family to isolate genes coding for new receptors. Using the hamster beta 2-adrenergic receptor gene as a hybridization probe we have isolated related genes including a cDNA encoding the rat D2 dopamine receptor. This receptor has been characterized on the basis of three criteria: the deduced amino-acid sequence which reveals that it is a member of the family of G-protein-coupled receptors; the tissue distribution of the mRNA which parallels that of the D2 dopamine receptor; and the pharmacological profile of mouse fibroblast cells transfected with the cDNA.

Repetitive, non-invasive imaging of the dopamine D2 receptor as a reporter gene in living animals

“…Construction of an adenovirus vector that expresses the dopamine D 2 receptor The rat D 2 R coding sequence 17 was cloned behind the cytomegalovirus (CMV) early promoter and inserted into the E1 region of a replication-deficient type 5 adenovirus to create Ad-D 2 R (Figure 1a). C6 rat glioma cells were infected with Ad-D 2 R or Ad-␤Gal, a control adenovirus expressing ␤-galactosidase.…”

“…FESP can be synthesized at high specific activity, has an IC 50 of 1.5 nm for the D 2 R (compared with 0.94 nm for spiperone), is currently used in humans, and -unlike most HSV1tk substrates -rapidly crosses the blood-brain barrier. [14][15][16] The D 2 R is a 415 amino acid protein with seven transmembrane domain topology 17,18 found in substantial levels primarily in the striatum and pituitary. 19 Because the D 2 R is encoded by an endogenous gene, ectopic expression as a reporter gene should not evoke an immune response.…”

Interactions of the Novel Antipsychotic Aripiprazole (OPC-14597) with Dopamine and Serotonin Receptor Subtypes

“…The D 2 -like receptors come from at least three genes and include multiple splice variants. The D 2 -like receptors [D 2S (Bunzow et al 1988), D 2L (Giros et al 1989;Monsma et al 1989), D 3 (Sokoloff et al 1990, and D 4 ] sometimes are linked to inhibition of cAMP synthesis and have a different pharmacological specificity from the D 1 -like receptors (i.e., having much higher affinity for spiperone or sulpiride).…”

“…The D 2 -like receptors come from at least three genes and include multiple splice variants. The D 2 -like receptors [D 2S (Bunzow et al 1988), D 2L (Giros et al 1989;Monsma et al 1989), D 3 (Sokoloff et al 1990, and D 4 ] sometimes are linked to inhibition of cAMP synthesis and have a different pharmacological specificity from the D 1 -like receptors (i.e., having much higher affinity for spiperone or sulpiride).The traditional view of antipsychotic drug efficacy posits a primary role for pharmacological antagonism of D 2 -like receptors. Despite the demonstrable effectiveness of dopamine D 2 receptor antagonists, however, a substantial number (up to 20%) of patients are considered unresponsive to these typical antipsychotics (Kane et al 1988).…”

Two forms of the rat D2 dopamine receptor as revealed by the polymerase chain reaction

“…To establish our own cell line expressing this dopamine D2 receptor, we designed oligonucleotide primers based on the published nucleotide sequence [2] in order to amplify rat striatal cDNA for the DZ receptor by the polymerase chain reaction (PCR) method [3]. By using PCR, we could circumvent cDNA library construction and screening and perhaps amplify other potentially interesting cDNAs which might share homology for the oligonucleotide primers.…”