Characterization of subtype-specific antibodies to the human D5 dopamine receptor: studies in primate brain and transfected mammalian cells.
To achieve a better understanding of how D5 dopamine receptors mediate the actions of dopamine in brain, we have developed antibodies specific for the D5 receptor. D5 antibodies reacted with recombinant baculovirus-infected Sf9 cells expressing the D5 receptor but not with the D1 receptor or a variety of other catecholaminergic and muscarinic receptors. Epitope-tagged D5 receptors expressed in mammalian cells were reactive with both D5 antibodies and an epitope-specific probe. A mixture of N-linked glycosylated polypeptides and higher molecular-mass species was detected on immunoblots of membrane fractions of D5-transfected cells and also of primate brain. D5 receptor antibodies intensely labeled pyramidal neurons in the prefrontal cortex, whereas spiny medium-sized neurons and aspiny large interneurons of the caudate nucleus were relatively lightly labeled. Antibodies to the D5 dopamine receptor should prove important in experimentally determining specific roles for the D5 and D1 receptors in cortical processes and diseases.
Bergson
C
C
Department of Pharmacology, Pennsylvania State College of Medicine, Milton S. Hershey Medical Center, Hershey 17033, USA.
Mrzljak
L
L
Lidow
M S
MS
Goldman-Rakic
P S
PS
Levenson
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D001923:Brain Chemistry; D002451:Cell Compartmentation; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D005625:Frontal Lobe / Q000033:anatomy & histology; D006801:Humans; D017403:In Situ Hybridization; D008253:Macaca mulatta; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000302:isolation & purification; D011954:Receptors, Dopamine / Q000145:classification / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D017447:Receptors, Dopamine D1; D050641:Receptors, Dopamine D5; D011994:Recombinant Proteins / Q000302:isolation & purification; D014018:Tissue Distribution
C493605:DRD5 protein, human; D000939:Epitopes; D012333:RNA, Messenger; D011954:Receptors, Dopamine; D017447:Receptors, Dopamine D1; D011994:Recombinant Proteins; D050641:Receptors, Dopamine D5