title: Cannabinoid receptor-interacting protein 1a modulates CB1 receptor signaling and regulation.
abstract: Cannabinoid CB1 receptors (CB1Rs) mediate the presynaptic effects of endocannabinoids in the central nervous system (CNS) and most behavioral effects of exogenous cannabinoids. Cannabinoid receptor-interacting protein 1a (CRIP1a) binds to the CB1R C-terminus and can attenuate constitutive CB1R-mediated inhibition of Ca(2+) channel activity. We now demonstrate cellular colocalization of CRIP1a at neuronal elements in the CNS and show that CRIP1a inhibits both constitutive and agonist-stimulated CB1R-mediated guanine nucleotide-binding regulatory protein (G-protein) activity. Stable overexpression of CRIP1a in human embryonic kidney (HEK)-293 cells stably expressing CB1Rs (CB1-HEK), or in N18TG2 cells endogenously expressing CB1Rs, decreased CB1R-mediated G-protein activation (measured by agonist-stimulated [(35)S]GTPγS (guanylyl-5'-[O-thio]-triphosphate) binding) in both cell lines and attenuated inverse agonism by rimonabant in CB1-HEK cells. Conversely, small-interfering RNA-mediated knockdown of CRIP1a in N18TG2 cells enhanced CB1R-mediated G-protein activation. These effects were not attributable to differences in CB1R expression or endocannabinoid tone because CB1R levels did not differ between cell lines varying in CRIP1a expression, and endocannabinoid levels were undetectable (CB1-HEK) or unchanged (N18TG2) by CRIP1a overexpression. In CB1-HEK cells, 4-hour pretreatment with cannabinoid agonists downregulated CB1Rs and desensitized agonist-stimulated [(35)S]GTPγS binding. CRIP1a overexpression attenuated CB1R downregulation without altering CB1R desensitization. Finally, in cultured autaptic hippocampal neurons, CRIP1a overexpression attenuated both depolarization-induced suppression of excitation and inhibition of excitatory synaptic activity induced by exogenous application of cannabinoid but not by adenosine A1 agonists. These results confirm that CRIP1a inhibits constitutive CB1R activity and demonstrate that CRIP1a can also inhibit agonist-stimulated CB1R signaling and downregulation of CB1Rs. Thus, CRIP1a appears to act as a broad negative regulator of CB1R function.
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Tricia H
authors: TH
authors: Smith
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Lawrence C
authors: LC
authors: Blume
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Alex
authors: A
authors: Straiker
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Jordan O
authors: JO
authors: Cox
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Bethany G
authors: BG
authors: David
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Julie R Secor
authors: JR
authors: McVoy
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Katherine W
authors: KW
authors: Sayers
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Justin L
authors: JL
authors: Poklis
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Rehab A
authors: RA
authors: Abdullah
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Michaela
authors: M
authors: Egertová
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Ching-Kang
authors: CK
authors: Chen
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Ken
authors: K
authors: Mackie
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Maurice R
authors: MR
authors: Elphick
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.).
authors: Allyn C
authors: AC
authors: Howlett
authors: Department of Pharmacology and Toxicology and Institute for Drug and Alcohol Studies (T.H.S., J.O.C., B.G.D., J.R.S.M., J.L.P., R.A.A., D.E.S.), Department of Anatomy and Neurobiology (K.W.S.), Department of Biochemistry and Molecular Biology (C.-K.C.), Virginia Commonwealth University School of Medicine, Richmond, Virginia; Department of Physiology and Pharmacology, Wake Forest University Health Sciences, Winston-Salem, North Carolina (L.C.B., A.C.H.); The Gill Center for Biomolecular Science and the Department of Psychological and Brain Sciences, Indiana University, Bloomington, Indiana (A.S., K.M.); and School of Biological and Chemical Sciences, Queen Mary, University of London, London, United Kingdom (M.E., M.R.E.) deselley@vcu.edu.
authors: Dana E
authors: DE
authors: Selley
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002531:Cerebellum / Q000378:metabolism; D063388:Endocannabinoids / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D009474:Neurons / Q000378:metabolism; D011869:Radioligand Assay; D017207:Rats, Sprague-Dawley; D043884:Receptor, Cannabinoid, CB1 / Q000819:agonists / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D002352:Carrier Proteins; C540799:Cnrip1 protein, rat; D063388:Endocannabinoids; D043884:Receptor, Cannabinoid, CB1; D019204:GTP-Binding Proteins
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