Deborah S. Hartman

RO-10-5824 is a selective dopamine D4 receptor agonist that increases novel object exploration in C57 mice

Novelty seeking as a behavioral phenomenon emerges as a compromise between approach and avoidance behavior. Although novelty seeking is thought to play a role in drug abuse and in cognition, the biological basis for this construct is poorly understood. At a genetic level, dopamine D4 receptors (D4R) appear to be critical for the behavioral expression of novelty seeking. In humans, polymorphisms of D4R have been associated with novelty-seeking traits in general and attention deficit-hyperactivity disorder in particular. Similarly, D4R (-/-) mice exhibit less novel object exploration than D4R (+/+) mice. Due to of the paucity of selective D4R ligands for use in behavioral pharmacology studies, few studies have examined the behavioral effects of D4R compounds in animals. The present experiments characterized RO-10-5824, a new, selective D4R partial agonist with minimal affinity for dopamine D2 and D3 receptors, and tested the hypothesis that activation of D4R increases the investigation by mice of a novel object placed in the center of a familiar open field. C57BL/6J and DBA/1J male mice were used in a dose response study of the selective D4R partial agonist RO-10-5824 (0, 1.0, 3.0, or 10.0 mg/kg). While having no effect on the amount of locomotor activity in novel or familiar environments, RO-10-5824 (10.0 mg/kg) increased time spent in the center of the enclosure in the presence of a novel object in C57 but not DBA mice. These results support the hypothesis that stimulation of D4R can enhance novelty seeking in mice and that this effect may be dependent on subtle genetic differences.

Dopamine receptor diversity: molecular and pharmacological perspectives.

Five distinct dopamine (DA) receptors, named D1-D5, have now been identified. All five DA receptors are expressed in the central nervous system where they control motor function, emotional states, and endocrine physiology, and all receptor subtypes are also found in peripheral tissues where they regulate transmembrane ion transport, catecholamine secretion, and cardiovascular function. Significant advances have been made following the cloning and heterologous expression of individual DA receptor subtypes, which has allowed the production of specific cDNA probes, receptor subtype-specific antibodies, and most recently the development of receptor-specific ligands. In this review we will discuss both molecular and pharmacological aspects of DA receptor diversity ranging from nucleotide sequences to behavioral aspects of DA receptor knockout mice.

Molecular attributes of dopamine receptors: new potential for antipsychotic drug development.

The neurotransmitter dopamine (DA) plays a central role in the control of motor function, emotional states, and endocrine physiology. The discovery that schizophrenic symptoms can be alleviated by neuroleptic drugs and the finding that these drugs interact at dopamine receptors has indicated involvement of the dopamine system in schizophrenia. The dopamine system has recently been shown to rely on the activation of five distinct subtypes of DA receptors (D1-D5) identified by molecular cloning, and pharmacological studies have specifically implicated the D2-like receptors (D2R, D3R and D4R) in antipsychotic action. In addition, the localization of D3R and D4R expression in the mesolimbic/mesocortical DA pathways is consistent with their proposed involvement in affective behaviour, and suggests that drugs developed specifically for these receptor subtypes might have potent antipsychotic activity with a lower propensity for extrapyramidal, endocrine, and cognitive side-effects.

Nerve Growth Factor-Induced Differentiation in Neuroblastoma Cells Expressing TrkA but Lacking p75NGFR

Abstract: Nerve growth factor (NGF) binds to two distinct cell surface receptors, TrkA, which is a receptor tyrosine kinase, and p75NGFR, whose role in NGF‐induced signal transduction remains unclear. We have found that human neuroblastoma IMR‐32 cells express TrkA, but p75NGFR expression was not detectable in these cells by northern blot analysis, immunoblotting, or chemical crosslinking experiments. Despite the lack of p75NGFR expression, subnanomolar concentrations of recombinant human NGF induced neurite outgrowth, tyrosine phosphorylation, and immediate early gene expression in these cells. These results strongly suggest that NGF‐induced neuronal differentiation in IMR‐32 cells is initiated through TrkA in the absence of p75NGFR. Thus, IMR‐32 cells may provide a model for studying neurotrophic effects of NGF on adult striatal cholinergic neurons, which also lack p75NGFR expression.

Expression and characterization of a dopamine D4R variant associated with delusional disorder

Multiple genetic polymorphisms of the human dopamine D4 receptor (hD4R) have been identified including a 12 bp repeat in exon 1 associated with a psychotic condition called delusional disorder. Competition binding assays revealed minor pharmacological differences between the recombinant A1 (normal) and A2 (delusional) proteins with respect to quinpirole and the antipsychotic clozapine, however no functional differences were detected for receptor activation by dopamine, epinephrine, or norepinephrine. Our results suggest that this polymorphism may only confer susceptibility to delusional disorder in combination with other genetic or environmental factors.

Development and characterization of antibodies against the N terminus of the human dopamine D4 receptor

Abstract: The human dopamine D4 receptor (hD4R), which has been implicated in human diseases such as schizophrenia and in a personality trait called “novelty seeking,” has not yet been characterized at the protein level. Following epitope scanning of the hD4R, we have produced a highly specific monoclonal antibody named DFR1 raised against an amino‐terminal peptide in a predicted extracellular region of the receptor. DFR1 decorated recombinant hD4Rs on the surface of intact Chinese hamster ovary (CHO) cells by flow cytometry and fluorescence microscopy and also recognized recombinant hD4.2, hD4.4, and hD4.7 receptor isoforms by western blot analysis. When expressed stably in CHO cells, all three hD4R isoforms contained N‐linked glycosylation and showed apparent molecular masses of 48, 55, and 67 kDa for hD4.2, hD4.4, and hD4.7, respectively. DFR1 immunoreactivity representing hD4R protein or dopamine D4 receptor‐like antigens was observed in crude membrane extracts of postmortem human brain tissue by immunoblotting. The DFR1 antibody provides a new immunological tool with the potential to further our understanding of the human dopamine D4 receptor protein.

Interaction of Antipsychotic Drugs with Dopamine Receptor Subtypes

Until the mid-twentieth century, patients suffering from psychiatric diseases such as schizophrenia were subjected to barbiturates, insulin coma, electroconvulsive therapy, and brain surgery, all of which provided little or no therapeutic benefit. The first major breakthrough in the treatment of psychiatric diseases occurred in the early 1950s with the discovery and introduction of chlorpromazine as the first effective neuroleptic (Delay et al. 1952). Improved neuroleptic drugs were subsequently developed, and the search began for the common “site of action” of these drugs.