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Regulation of native GABAA receptors by PKC and protein phosphatase activity

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Abstract

Rationale and objective

Protein kinase C (PKC) modulation of ionotropic receptors is a common mechanism for regulation of channel function. The effects of PKC and phosphatase activation on native gamma-aminobutyric acid (GABAA) receptors in adult brain are unknown. Previous studies of recombinant GABAA receptors have provided evidence that PKC activation inhibits receptor function, whereas other studies suggest that PKC either increases or does not alter GABAA receptor function. The present study explored (a) the effects of PKC and phosphatase activity on GABA-mediated 36Cl− uptake in cerebral cortical synaptoneurosomes and (b) the effect of PKC activity on muscimol-induced loss of righting reflex (LORR) in adult rats.

Methods

GABAA receptor function in vitro was measured by muscimol-induced 36Cl− uptake into cerebral cortical synaptoneurosomes. The in vivo effect of PKC on GABAA-mediated function was measured by intracerebroventricular (i.c.v.) injection of 4-beta-phorbol-12,13-dibutyrate (PDBu) or calphostin C followed by determination of muscimol-induced LORR.

Results

Adenosine triphosphate (ATP) and PDBu produced a concentration-dependent and specific reduction in muscimol-stimulated 36Cl− uptake that was blocked by the PKC inhibitor calphostin C. Both adenosine diphosphate and 4αPDBu were ineffective. Phosphatase inhibition produced similar inhibition of muscimol responses. Furthermore, i.c.v. administration of PDBu and calphostin C produced opposing effects on both the onset and the duration of muscimol-induced LORR in rats.

Conclusions

The present study provides evidence that PKC activation reduces GABAA receptor function in native receptors both in vitro and in vivo. Phosphatase inhibitors decrease muscimol-mediated Cl− uptake in GABAA receptors demonstrating coordinated regulation of native receptors by PKC and phosphatases.

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References

  • Battastini AM, da Rocha JB, Barcellos CK, Dias RD, Sarkis JJ (1991) Characterization of an ATP diphosphohydrolase (EC 3.6.1.5) in synaptosomes from cerebral cortex of adult rats. Neurochem Res 16:1303–1310

    Article  PubMed  CAS  Google Scholar 

  • Boehm SL II, Peden L, Harris RA, Blednov Y (2004) Deletion of the fyn-kinase gene alters sensitivity to GABAergic drugs: dependence on β2/β3 GABAA receptor subunits. J Pharmacol Exp Ther 309:1154–1159

    Article  PubMed  CAS  Google Scholar 

  • Bowers BJ, Wehner JM (2001) Mice lacking PKCg exhibit increased ethanol consumption and behavioral impulsivity. Alcohol Clin Exp Res 25:119A

    Google Scholar 

  • Brandon NJ, Delmas P, Kittler JT, McDonald BJ, Sieghart W, Brown DA, Smart TG, Moss SJ (2000) GABAA receptor phosphorylation and functional modulation in cortical neurons by a protein kinase C-dependent pathway. J Biol Chem 275:38856–38862

    Article  PubMed  CAS  Google Scholar 

  • Chander A, Sen N, Wu AM, Spitzer AR (1995) Protein kinase C in ATP regulation of lung surfactant secretion in type II cells. Am J Physiol 268:L108–16

    PubMed  CAS  Google Scholar 

  • Chapell R, Bueno OF, Alvarez-Hernandez X, Robinson LC, Leidenheimer NJ (1998) Activation of protein kinase C induces gamma-aminobutyric acid type A receptor internalization in Xenopus oocytes. J Biol Chem 273:32595–32601

    Article  PubMed  CAS  Google Scholar 

  • Chen ZW, Chang CS, Leil TA, Olcese R, Olsen RW (2005) GABARAP regulates GABAA receptor cell surface number in Xenopus laevis oocytes. Mol Pharmacol 68(1):152–159

    Article  PubMed  CAS  Google Scholar 

  • Cooley RK, Vanderwolf CH (1990) Stereotaxic surgery in the rat: a photographic series. A J Kirby Co., London, Canada

    Google Scholar 

  • Feng J, Cai X, Zhao J, Yan Z (2001) Serotonin receptors modulate GABA(A) receptor channels through activation of anchored protein kinase C in prefrontal cortical neurons. J Neurosci 21:6502–6511

    PubMed  CAS  Google Scholar 

  • Flores-Hernandez J, Hernandez S, Snyder GL, Yan Z, Fienberg AA, Moss SJ, Greengard P, Surmeier DJ (2000) D(1) dopamine receptor activation reduces GABA(A) receptor currents in neostriatal neurons through a PKA/DARPP-32/PP1 signaling cascade. J Neurophysiol 83:2996–3004

    PubMed  CAS  Google Scholar 

  • Fritschy JM, Brunig I (2003) Formation and plasticity of GABAergic synapses: physiological mechanisms and pathophysiological implications. Pharmacol Ther 98:299–323

    Article  PubMed  CAS  Google Scholar 

  • Ghansah E, Weiss DS (2001) Modulation of GABA(A) receptors by benzodiazepines and barbiturates is autonomous of PKC activation. Neuropharmacology 40:327–333

    Article  PubMed  CAS  Google Scholar 

  • Harris RA, Allan AM (1985) Functional coupling of GABA receptors to chloride channels in brain membranes. Science 228:1108–1110

    Article  PubMed  CAS  Google Scholar 

  • Harris RA, McQuilkin SJ, Paylor R, Abeliovich A, Tonegawa S, Wehner JM (1995) Mutant mice lacking the gamma isoform of protein kinase C show decreased behavioral actions of ethanol and altered function of gamma-aminobutyrate type A receptors. Proc Natl Acad Sci U S A 92:3658–3662

    Article  PubMed  CAS  Google Scholar 

  • Herring D, Huang R, Singh M, Dillon GH, Leidenheimer NJ (2005) PKC modulation of GABAA receptor endocytosis and function is inhibited by mutation of a dileucine motif within the receptor beta 2 subunit. Neuropharmacology 48:181–194

    Article  PubMed  CAS  Google Scholar 

  • Hodge CW, Mehmert KK, Kelley SP, McMahon T, Haywood A, Olive MF, Wang D, Sanchez-Perez AM, Messing RO (1999) Supersensitivity to allosteric GABAA receptor modulators and alcohol in mice lacking PKCe. Nat Neurosci 2:997–1002

    Article  PubMed  CAS  Google Scholar 

  • Hodge CW, Raber J, McMahon T, Walter H, Sanchez-Perez AM, Olive MF, Mehmert K, Morrow AL, Messing RO (2002) Decreased anxiety-like behavior, reduced stress hormones, and neurosteroid supersensitivity in mice lacking protein kinase CE. J Clin Invest 110:1003–1010

    Article  PubMed  CAS  Google Scholar 

  • James S, Richardson PJ (1993) Production of adenosine from extracellular ATP at the striatal cholinergic synapse. J Neurochem 60:219–227

    Article  PubMed  CAS  Google Scholar 

  • Kapur J, Haas KF, Macdonald RL (1999) Physiological properties of GABAA receptors from acutely dissociated rat dentate granule cells. J Neurophysiol 81:2464–2471

    PubMed  CAS  Google Scholar 

  • Kittler JT, Delmas P, Jovanovic JN, Brown DA, Smart TG, Moss SJ (2000) Constitutive endocytosis of GABAA receptors by an association with the adaptin AP2 complex modulates inhibitory synaptic currents in hippocampal neurons. J Neurosci 20:7972–7977

    PubMed  CAS  Google Scholar 

  • Kralic JE, Homanics GE, Morrow AL (2001) Knockout of the GABAA receptor alpha1 subunit alters receptor function and pharmacology. Genes & gene delivery for diseases of alcoholism international symposium 1:39

    Google Scholar 

  • Kralic JE, Korpi ER, O'Buckley TK, Homanics GE, Morrow AL (2002a) Molecular and pharmacological characterization of GABAA receptor a1 subunit knockout mice. J Pharmacol Exp Ther 302:1037–1045

    Article  PubMed  CAS  Google Scholar 

  • Kralic JE, O'Buckley TK, Khisti RT, Hodge CW, Homanics GE, Morrow AL (2002b) GABAA receptor a1 subunit deletion alters receptor subtype assembly, pharmacological and behavioral responses to benzodiazepines and zolpidem. Neuropharmacology 43:685–694

    Article  PubMed  CAS  Google Scholar 

  • Krupp J, Feltz P (1993) Synaptic- and agonist-induced chloride currents in neonatal rat sympathetic preganglionic neurones in vitro. J Physiol 471:729–748

    PubMed  CAS  Google Scholar 

  • Leidenheimer NJ, McQuiklin SJ, Hahner LD, Whiting P, Harris RA (1992) Activation of protein kinase C selectively inhibits the λ-aminobutyric acidA receptor: role of desensitization. Mol Pharmacol 41:1116–1123

    PubMed  CAS  Google Scholar 

  • Leil TA, Chen ZW, Chang CS, Olsen RW (2004) GABAA receptor-associated protein traffics GABAA receptors to the plasma membrane in neurons. J Neurosci 24:11429–11438

    Article  PubMed  CAS  Google Scholar 

  • Lin Y-F, Browning MD, Dudek EM, Macdonald RL (1994) Protein kinase C enhances recombinant bovine α1β1gamma2L GABAA receptor whole-cell currents expressed in L929 fibroblasts. Neuron 13:1421–1431

    Article  PubMed  CAS  Google Scholar 

  • Morrow AL, Suzdak PD, Karanian JW, Paul SM (1988) Chronic ethanol administration alters λ-aminobutyric acid, pentobarbital and ethanol-mediated 36Cl− uptake in cerebral cortical synaptoneurosomes. J Pharmacol Exp Ther 246:158–164

    PubMed  CAS  Google Scholar 

  • Moss SJ, Smart TG (2001) Constructing inhibitory synapses. Nat Rev Neurosci 2:240–250

    Article  PubMed  CAS  Google Scholar 

  • Oh S, Jang CG, Ma T, Ho IK (1999) Activation of protein kinase C by phorbol dibutyrate modulates GABAA receptor binding in rat brain slices. Brain Res 850:158–165

    Article  PubMed  CAS  Google Scholar 

  • Paxinos G, Watson C (1998) The rat brain. Academic Press, New York

    Google Scholar 

  • Poisbeau P, Cheney MC, Browning MD, Mody I (1999) Modulation of synaptic GABAA receptor function by PKA and PKC in adult hippocampal neurons. J Neurosci 19:674–683

    PubMed  CAS  Google Scholar 

  • Proctor WR, Poelchen W, Bowers BJ, Wehner JM, Messing RO, Dunwiddie TV (2003) Ethanol differentially enhances hippocampal GABA A receptor-mediated responses in protein kinase C gamma (PKC gamma) and PKC epsilon null mice. J Pharmacol Exp Ther 305:264–270

    Article  PubMed  CAS  Google Scholar 

  • Rico EP, Senger MR, Fauth Mda G, Dias RD, Bogo MR, Bonan CD (2003) ATP and ADP hydrolysis in brain membranes of zebrafish (Danio rerio). Life Sci 73:2071–2082

    Article  PubMed  CAS  Google Scholar 

  • Robello M, Amico C, Cupello A (1997) A dual mechanism for impairment of GABAA receptor activity by NMDA receptor activation in rat cerebellum granule cells. Eur Biophys J 25:181–187

    Article  PubMed  CAS  Google Scholar 

  • Sassoe-Pognetto M, Panzanelli P, Sieghart W, Fritschy JM (2000) Colocalization of multiple GABA(A) receptor subtypes with gephyrin at postsynaptic sites. J Comp Neurol 420:481–498

    Article  PubMed  CAS  Google Scholar 

  • Schwartz RD, Suzdak PD, Paul SM (1986) λ-aminobutyric acid (GABA) and barbiturate receptor-mediated 36Cl− uptake in rat brain synaptoneurosomes: evidence for rapid desensitization of the GABA receptor-coupled chloride ion channel. Mol Pharmacol 30:419–426

    PubMed  CAS  Google Scholar 

  • Sieghart W (2000) Unraveling the function of GABAA receptor subtypes. Trends Pharmacol Sci 21:411–413

    Article  PubMed  CAS  Google Scholar 

  • Sokolova E, Nistri A, Giniatullin R (2001) Negative cross talk between anionic GABAA and cationic P2X ionotropic receptors of rat dorsal root ganglion neurons. J Neurosci 21:4958–4968

    PubMed  CAS  Google Scholar 

  • Ticku MK, Mehta AK (1990) gamma-Aminobutyric acidA receptor desensitization in mice spinal cord cultured neurons: lack of involvement of protein kinases A and C. Mol Pharmacol 38:719–724

    PubMed  CAS  Google Scholar 

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Acknowledgements

The authors thank Todd O'Buckley for technical assistance and Dr. Jason Kralic and Dr. Clyde Hodge for helpful discussions. This work was supported by National Institutes of Health grants AA015409 (SK) and AA11605 (ALM).

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Authors and Affiliations

  1. Departments of Psychiatry and Pharmacology, Bowles Center for Alcohol Studies, UNC School of Medicine, Chapel Hill, NC, 27599, USA

    Sandeep Kumar, Rahul T. Khisti & A. Leslie Morrow

  2. Center for Alcohol Studies, CB#7178, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599-7178, USA

    A. Leslie Morrow

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  1. Sandeep Kumar
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  2. Rahul T. Khisti
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  3. A. Leslie Morrow
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Correspondence to A. Leslie Morrow.

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Kumar, S., Khisti, R.T. & Morrow, A.L. Regulation of native GABAA receptors by PKC and protein phosphatase activity. Psychopharmacology 183, 241–247 (2005). https://doi.org/10.1007/s00213-005-0161-x

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  • DOI: https://doi.org/10.1007/s00213-005-0161-x

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