Rebecca Stadel
University Of Connecticut Storrs
Project start date: 2010-01-15
Project end date: 2013-01-14
Sponsored Links Excellgen http://Excellgen.com
Rebecca Stadel
University Of Connecticut Storrs
Project start date: 2010-01-15
Project end date: 2013-01-14
Grants awarded to Rebecca Stadel
CANNABINOID RECEPTOR ONE-B ARRESTIN INTERACTIONS
Rebecca Stadel, Postdoctoral Fellow
University Of Connecticut Storrs, 438 Whitney Road Extension, Unit 1133, Storrs-mansfield, Ct 06269
Grant 1F32DA028080-01 from National Institute On Drug Abuse
Abstract: The cannabinoid receptor one (CB1) is a G protein-coupled receptor (GPCR) that binds the main psychoactive component of marijuana, A9-tetrahydrocannabinol. The CB1 receptor exerts its effects by binding to Gai/o G-proteins, resulting in the activation of effector pathways. Desensitization and internalization of the receptor govern the rate and magnitude of CB1 receptor activation. Both processes, for most non-visual GPCRs, are initiated following (3-arrestin binding, yet the mechanisms of a (3-arrestin-CB1 interaction, as well as the consequences on receptor trafficking, are poorly understood. The overall objectives of this project is to determine the p-arrestin subtype(s) that bind to the CB1 receptor, identify critical residues and structural features of the CB1 receptor that are involved in p-arrestin binding, and examine the cellular consequences of such an interaction. In Aim 1 of this proposal, the structural requirements of the CB1 receptor that are critical for binding purified P-arrestin(s) and the optimal binding environment will be delineated, using a combination of biochemical and NMR studies. Affinities of both p- arrestin subtypes for the CB1 receptor will be measured and will elucidate the preferred binding partner. To examine the consequences of CB1-p-arrestin interactions, in Aim 2 the subcellular distribution of p-arrestin and the full-length CB1 receptor will be examined individually and when co-expressed, in HEK293 and primary hippocampal neurons using confocal microscopy. In Aim 3, the role of structural motifs of the CB1 receptor critical for P-arrestin binding (identified in Aim 1) will be assessed in the full-length receptor using targeted mutatgenesis and the consequences of these mutations on receptor trafficking examined. These studies will improve our understandings of the mechanisms involved in desensitization and internalization of cannabinoid receptors, and by elucidating the exact nature of the CB1-P-arrestin complex, may elucidate novel drug targets. Relevance The cannabinoid receptor one (CB1) is primarily distributed in the central nervous system. Agonist activation of the CB1 receptor has been shown to cause an increase in appetite, produce anxiety, sedation, and hypothermia, as well as be detrimental to learning and memory, whereas inverse agonist blockade results in appetite suppression and a decreased craving for drugs. The goal of this project is to elucidate CB1- beta arrestin interactions that lead to a functional decrease in CB1 and consequently impact the availability of receptors for therapeutic strategies
Keywords: 6H-Dibenzo(b, d)pyran-1-ol, 6a, 7, 8, 10a-tetrahydro-6, 6, 9-trimethyl-3-pentyl-, (6aR-trans)-; 9-ene-Tetrahydrocannabinol; Absence of pain sensation; Absence of sensibility to pain; Adverse effects; Affinity; Agonist; Amino Acids; Ammon Horn; Anxiety; Appetite; Arrestins; Assay; Attenuated; Binding; Binding (Molecular Function); Bioassay; Biochemical; Biologic Assays; Biological Assay; Body Weight decreased; CB1 Receptor; Cannabinoids, Endogenous; Cell Line; Cell Lines, Strains; CellLine; Cells; Central Nervous System; Cessation of smoking; Chemicals; Classification; Co-Immunoprecipitations; Complex; Confocal Microscopy; Cornu Ammonis; Delta-9-Tetrahydrocannabinol; Desire for food; Development; Dronabinol; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Drug abuse; Endocannabinoids; Environment; Event; Family; Feels no pain; G Protein Go; G Protein-Complex Receptor; G(o) Protein; G-Protein, Go Subunit; G-Protein, Inhibitory Go; G-Protein-Coupled Receptors; GTP Binding Protein alpha Subunit, Go; Genetic Alteration; Genetic Change; Genetic defect; Genetics-Mutagenesis; Go Alpha Subunit; Goals; Guanine Nucleotide-Binding Protein Go; Hippocampus; Hippocampus (Brain); Hypothermia; In Vitro; Individual; Label; Lead; Learning; Length; Link; Marihuana; Marinol; Measures; Memory; Microscopy, Confocal; Molecular Biology, Mutagenesis; Molecular Interaction; Mutagenesis; Mutation; Nature; Nerve Cells; Nerve Unit; Nervous System, CNS; Neural Cell; Neuraxis; Neurocyte; Neurons; No sensitivity to pain; Nuclear Magnetic Resonance; Pathway interactions; Pb element; Process; Receptor Activation; Receptor Protein; Receptor, Cannabinoid, CB1; Relative; Relative (related person); Role; Sedation procedure; Site; Structure; Systematics; Tetrahydrocannabinol; Therapeutic; Time; Treatment Side Effects; Visual; Weight Loss; Weight Reduction; abuse of drugs; abuses drugs; aminoacid; analgesia; arr3; arrestin 1; arrestin 2; arrestin 3; arrestin B; arrestin3; beta-arrestin; body weight loss; cannabinoid receptor; cease smoking; cultured cell line; delta(1)-THC; delta(1)-Tetrahydrocannabinol; delta(9)-THC; delta(9)-Tetrahydrocannabinol; depression; desensitization; drug craving; experiment; experimental research; experimental study; genome mutation; heavy metal Pb; heavy metal lead; hippocampal; hypothermia, natural; improved; in vivo; interest; natural hypothermia; neuronal; novel; pathway; prevent; preventing; receptor; receptor binding; receptor internalization; research study; sedation; side effect; smoking cessation; social role; therapy adverse effect; trafficking; treatment adverse effect; wt-loss
Project start date: 2010-01-15
Project end date: 2013-01-14
Budget start date: 15-JAN-2010
Budget end date: 14-JAN-2011
1F32DA028080-01 (2010): $47604