Distribution of D4 dopamine receptor in rat brain with sequence-specific antibodies

被引:128
作者
Defagot, MC
Malchiodi, EL
Villar, MJ
Antonelli, MC
机构
[1] UNIV BUENOS AIRES,CONICET,INST QUIM & FISICOQUIM BIOL,FAC FARM & BIOQUIM,RA-1113 BUENOS AIRES,DF,ARGENTINA
[2] UNIV BUENOS AIRES,FAC FARM & BIOQUIM,CATEDRA INMUNOL,RA-1113 BUENOS AIRES,DF,ARGENTINA
[3] INST NEUROBIOL,BUENOS AIRES,DF,ARGENTINA
来源
MOLECULAR BRAIN RESEARCH | 1997年 / 45卷 / 01期
关键词
D4 dopaminergic receptor; antipeptide antibody; benzamide binding; immunocytochemistry; rat brain;
D O I
10.1016/S0169-328X(96)00235-5
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The distribution of the dopaminergic D4 receptor in rat brain was studied employing site directed polyclonal antibodies. Antisera were raised in rabbits to two oligopeptides corresponding to amino acids 160-172 of the second extracellular loop (Pi) and amino acids 260-273 of the third intracellular loop (P2) of the D4 receptor sequence. Affinity-purified antibodies (anti-P1 and anti-P2) specifically recognized two major bands of 42-45 and 95 kDa in Westen blots of denatured preparations of various rat brain areas. Immunocytochemistry studies showed that D4 receptor is widely distributed in rat central nervous system (CNS) showing higher labelling in the hippocampus (CA1, CA2, CA3 and dentate gyrus) frontal cortex, entorhinal cortex, caudate putamen, nucleus accumbens, olfactory tubercle, cerebellum, supraoptic nucleus and sustancia nigra pars compacta. In addition, anti-pi decreased the binding of the antagonist [H-3]YR4-09151-2 selective for D2, D3 and D4 receptors but did not modify the binding of [3H]raclopride an antagonist selective for D2 and D3, in striatal synaptosomes. Anti-P2 did not modify the binding of these ligands. These results confirm the selectivity of the antibodies towards the D4 receptor and suggest that the binding site for the antagonists might be located at or close to the second extracellular loop of the protein sequence. D4 receptor protein is mainly expressed in plasma membranes and in the peripheral cytoplasm of neurons and is more widely distributed than was originally proposed based on mRNA localization, since it is present both in limbic, diencephalic and motor areas of rat brain.
引用
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页码:1 / 12
页数:12
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