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Supersensitivity to allosteric GABAA receptor modulators and alcohol in mice lacking PKCε

Nature Neuroscience volume 2, pages 997–1002 (1999)Cite this article

Abstract

Several of the actions of ethanol are mediated by γ-aminobutyrate type A (GABAA) receptors. Here we demonstrated that mutant mice lacking protein kinase C epsilon (PKCε) were more sensitive than wild-type littermates to the acute behavioral effects of ethanol and other drugs that allosterically activate GABAA receptors. GABAA receptors in membranes isolated from the frontal cortex of PKCε null mice were also supersensitive to allosteric activation by ethanol and flunitrazepam. In addition, these mutant mice showed markedly reduced ethanol self-administration. These findings indicate that inhibition of PKCε increases sensitivity of GABAA receptors to ethanol and allosteric modulators. Pharmacological agents that inhibit PKCε may be useful for treatment of alcoholism and may provide a non-sedating alternative for enhancing GABAA receptor function to treat other disorders such as anxiety and epilepsy.

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Figure 1: Reduced alcohol intake and preference and normal consummatory behavior by PKCε–/– and PKCε+/+ mice.
Figure 2: Locomotor activity and acute responses to ethanol by PKCε–/– and PKCε+/+ mice.
Figure 3: Acute sedative and locomotor activating effects of GABAA receptor modulators in PKCε–/– and PKCε+/+ mice.
Figure 4: Muscimol-stimulated 36Cl– uptake in cortical microsacs.

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References

  1. Hodge, C., Chappelle, A. & Samson, H. GABAergic transmission in the nucleus accumbens is involved in the termination of ethanol self-administration in rats. Alcohol Clin. Exp. Res. 19, 1486–1493 (1995).

    Article  CAS  Google Scholar 

  2. Hodge, C. W. & Cox, A. A. The discriminative stimulus effects of ethanol are mediated by NMDA and GABA(A) receptors in specific limbic brain regions. Psychopharmacology 139, 95–107 (1998).

    Article  CAS  Google Scholar 

  3. Dietrich, R., Dunwiddie, T., Harris, R. & Erwin, V. Mechanism of action of ethanol: initial central nervous system actions. Pharmacol. Rev. 41, 489–537 (1989).

    Google Scholar 

  4. Allan, A. & Harris, R. Acute and chronic ethanol treatments alter GABA receptor-operated chloride channels. Pharmacol. Biochem. Behav. 27, 665–670 (1987).

    Article  CAS  Google Scholar 

  5. Mehta, A. K. & Ticku, M. K. Ethanol potentiation of GABAergic transmission in cultured spinal cord neurons involves γ-aminobutyric acidA-gated chloride channels. J. Pharmacol. Exp. Ther. 246, 558–564 (1988).

    CAS  PubMed  Google Scholar 

  6. Suzdak, P. D., Schwartz, R. D., Skolnick, P. & Paul, S. M. Ethanol stimulates γ-aminobutyric acid receptor-mediated chloride transport in rat brain synaptoneurosomes. Proc. Natl. Acad. Sci. USA 83, 4071–4075 (1986).

    Article  CAS  Google Scholar 

  7. Siggins, G. R., Pittman, Q. J. & French, E. D. Effects of ethanol on CA1 and CA3 pyramidal cells in the hippocampal slice preparation: an intracellular study. Brain Res. 414, 22–34 (1987).

    Article  CAS  Google Scholar 

  8. Givens, B. S. & Breese, G. R. Site-specific enhancement of gamma-aminobutyric acid-mediated inhibition of neural activity by ethanol in the rat medial septal area. J. Pharmacol. Exp. Ther. 254, 528–538 (1990).

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Proctor, W. R., Soldo, B. L., Allan, A. M. & Dunwiddie, T. V. Ethanol enhances synaptically evoked GABAA receptor-mediated responses in cerebral cortical neurons in rat brain slices. Brain Res. 595, 220–227 (1992).

    Article  CAS  Google Scholar 

  10. Weiner, J. L., Zhang, L. & Carlen, P. L. Potentiation of GABAA -mediated synaptic current by ethanol in hippocampal CA1 neurons: Possible role of protein kinase C. J. Pharmacol. Exp. Ther. 268, 1388–1395 (1994).

    CAS  PubMed  Google Scholar 

  11. Weiner, J., Valenzuela, C., Watson, P., Frazier, C. & Dunwiddie, T. Elevation of basal protein kinase C activity increases ethanol sensitivity of GABA A receptors in rat hippocampal CA1 pyramidal neurons. J. Neurochem. 68, 1949–1959 (1997).

    Article  CAS  Google Scholar 

  12. Moss, S. J. & Smart, T. G. Modulation of amino acid-gated ion channels by protein phosphorylation. Int. Rev. Neurobiol. 39, 1–52 (1996).

    Article  CAS  Google Scholar 

  13. Wafford, K. A. et al. Ethanol sensitivity of the GABAA receptor expressed in Xenopus oocytes requires eight amino acids contained in the γ2L subunit of the receptor complex. Neuron 7, 27–33 (1991).

    Article  CAS  Google Scholar 

  14. Marszalec, W., Kurata, Y., Hamilton, B. J., Carter, D. B. & Narahashi, T. Selective effects of alcohols on gamma-aminobutyric acid A receptor subunits expressed in human embryonic kidney cells. J. Pharmacol. Exp. Ther. 269, 157–163 (1994).

    CAS  PubMed  Google Scholar 

  15. Sigel, E., Baur, R. & Malherbe, P. Recombinant GABAA receptor function and ethanol. FEBS Lett. 324, 140–142 (1993).

    Article  CAS  Google Scholar 

  16. Sapp, D. W. & Yeh, H. H. Ethanol-GABAA receptor interactions: a comparison between cell lines and cerebellar Purkinje cells. J. Pharmacol. Exp. Ther. 284, 768–776 (1998).

    CAS  PubMed  Google Scholar 

  17. Homanics, G. E. et al. Normal electrophysiological and behavioral responses to ethanol in mice lacking the long splice variant of the gamma2 subunit of the gamma-aminobutyrate type A receptor. Neuropharmacology 38, 253–265 (1999).

    Article  CAS  Google Scholar 

  18. Tanaka, C. & Nishizuka, Y. The protein kinase C family for neuronal signaling. Annu. Rev. Neurosci. 17, 551–567 (1994).

    Article  CAS  Google Scholar 

  19. Saito, N. et al. Cellular and intracellular localization of ε-subspecies of protein kinase C in the rat brain; presynaptic localization of the ε-subspecies. Brain Res. 607, 241–248 (1993).

    Article  CAS  Google Scholar 

  20. Gordon, A. S., Yao, L., Wu, Z. L., Coe, I. R. & Diamond, I. Ethanol alters the subcellular localization of delta- and epsilon protein kinase C in NG108-15 cells. Mol. Pharmacol. 52, 554–559 (1997).

    Article  CAS  Google Scholar 

  21. Messing, R. O., Petersen, P. J. & Henrich, C. J. Chronic ethanol exposure increases levels of protein kinase C delta and epsilon and protein kinase C-mediated phosphorylation in cultured neural cells. J. Biol. Chem. 266, 23428–23432 (1991).

    CAS  PubMed  Google Scholar 

  22. Coe, I. R., Yao, L., Diamond, I. & Gordon, A. S. The role of protein kinase C in cellular tolerance to ethanol. J. Biol. Chem. 271, 29468–29472 (1996).

    Article  CAS  Google Scholar 

  23. Hundle, B. et al. An inhibitory fragment derived from protein kinase C epsilon prevents enhancement of nerve growth factor responses by ethanol and phorbol esters. J. Biol. Chem. 272, 15028–15035 (1997).

    Article  CAS  Google Scholar 

  24. Hodge, C., Slawecki, C. & Aiken, A. Norepinephrine and serotonin receptors in the paraventricular nucleus of the hypothalamus interactively modulate ethanol consumption. Alcohol. Clin. Exp. Res. 21, 250–260 (1996).

    Google Scholar 

  25. Phillips, T. J. et al. Alcohol preference and sensitivity are markedly reduced in mice lacking dopamine D2 receptors. Nat. Neurosci. 1, 610–615 (1998).

    Article  CAS  Google Scholar 

  26. Crabbe, J. C. et al. Elevated alcohol consumption in null mutant mice lacking 5-HT1B serotonin receptors. Nat. Genet. 14, 98–101 (1996).

    Article  CAS  Google Scholar 

  27. Schuckit, M. A. Low level of response to alcohol as a predictor of future alcoholism. Am. J. Psychiatry 151, 184–189 (1994).

    Article  CAS  Google Scholar 

  28. Thiele, T. E., Marsh, D. J., Ste. Marie, L., Bernstein, I. L. & Palmiter, R. D. Ethanol consumption and resistance are inversely related to neuropeptide Y levels. Nature 396, 366–369 (1998).

    Article  CAS  Google Scholar 

  29. Finn, D. A., Syapin, P. J., Bejanian, M., Jones, B. L. & Alkana, R. L. Temperature dependence of ethanol depression in mice: dose response. Alcohol. Clin. Exp. Res. 18, 382–386 (1994).

    Article  CAS  Google Scholar 

  30. Frye, G. D. & Breese, G. R. An evaluation of the locomotor stimulating action of ethanol in rats and mice. Psychopharmacology 75, 372–379 (1981).

    Article  CAS  Google Scholar 

  31. Miyakawa, T. et al. Fyn-kinase as a determinant of ethanol sensitivity: relation to NMDA-receptor function. Science 278, 698–701 (1997).

    Article  CAS  Google Scholar 

  32. Deitrich, R. A., Dunwiddie, T. V., Harris, R. A. & Erwin, V. G. Mechanism of action of ethanol: Initial central nervous system actions. Pharmacol. Rev. 41, 489–537 (1989).

    CAS  PubMed  Google Scholar 

  33. Lovinger, D. M., White, G. & Weight, F. Ethanol inhibits NMDA-activated ion current in hippocampal neurons. Science 243, 1721–1724 (1989).

    Article  CAS  Google Scholar 

  34. Guidotti, A., Corda, M. G., Wise, B. C., Vaccarino, F. & Costa, E. GABAergic synapses. Supramolecular organization and biochemical regulation. Neuropharmacology 22, 1471–1479 (1983).

    Article  CAS  Google Scholar 

  35. Olsen, R. W. Drug interactions at the GABA receptor-ionophore complex. Annu. Rev. Pharmacol. Toxicol. 22, 245–277 (1982).

    Article  CAS  Google Scholar 

  36. Johnson, J. A., Gray, M. O., Chen, C.-H. & Mochly-Rosen, D. A protein kinase C translocation inhibitor as isozyme-specific antagonist of cardiac function. J. Biol. Chem. 271, 24962–24966 (1996).

    Article  CAS  Google Scholar 

  37. Harris, R. A. et al. Mutant mice lacking the γ isoform of protein kinase C show decreased behavioral actions of ethanol and altered function of γ-aminobutyrate type A receptors. Proc. Natl. Acad. Sci. USA 92, 3658–3662 (1995).

    Article  CAS  Google Scholar 

  38. Kellenberger, S., Malherbe, P. & Sigel, E. Function of the α1β2γ2S γ-aminobutyric acid type A receptor is modulated by protein kinase C via multiple phosphorylation sites. J. Biol. Chem. 267, 25660–25663 (1992).

    CAS  PubMed  Google Scholar 

  39. Krishek, B. J. et al. Regulation of GABAA receptor function by protein kinase C phosphorylation. Neuron 12, 1081–1095 (1994).

    Article  CAS  Google Scholar 

  40. Leidenheimer, N. J. & Harris, R. A. Acute effects of ethanol on GABAA receptor function: molecular and physiological determinants. Adv. Biochem. Psychopharmacol. 47, 269–279 (1992).

    CAS  PubMed  Google Scholar 

  41. Lin, Y. F., Browning, M. D., Dudek, E. M. & Macdonald, R. L. Protein kinase C enhances recombinant bovine α1β1γ2L GABAA receptor whole-cell currents expressed in L929 fibroblasts. Neuron 13, 1421–1431 (1994).

    Article  CAS  Google Scholar 

  42. Lin, Y. F., Angelotti, T. P., Dudek, E. M., Browning, M. D. & Macdonald, R. L. Enhancement of recombinant alpha 1 beta 1 gamma 2L gamma-aminobutyric acidA receptor whole-cell currents by protein kinase C is mediated through phosphorylation of both beta 1 and gamma 2L subunits. Mol. Pharmacol. 50, 185–195 (1996).

    CAS  PubMed  Google Scholar 

  43. Poisbeau, P., Cheney, M. C., Browning, M. D. & Mody, I. Modulation of synaptic GABAA receptor function by PKA and PKC in adult hippocampal neurons. J. Neurosci. 19, 674–683 (1999).

    Article  CAS  Google Scholar 

  44. Mochly-Rosen, D. & Gordon, A. Anchoring proteins for protein kinase C: a means for isozyme selectivity. FASEB J. 12, 35–42 (1998).

    Article  CAS  Google Scholar 

  45. Khasar, S. et al. A novel nociceptive signaling pathway demonstrated in PKCe-mutant mice. Neuron (in press).

  46. Leidenheimer, N. J., McQuilkin, S. J., Hahner, L. D., Whiting, P. & Harris, R. A. Activation of protein kinase C selectively inhibits the γ-aminobutyric acidA receptor: role of desensitization. Mol. Pharmacol. 41, 1116–1123 (1992).

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by funds from the State of California for Alcohol and Drug Abuse Research to C.W.H. and R.O.M. and by a grant from the Alcoholic Beverage Medical Research Foundation to C.W.H.

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  1. Department of Neurology and Ernest Gallo Clinic and Research Center, University of California San Francisco, 1001 Potrero Avenue; Building 1, Room 101, San Francisco, 94110, California, USA

    Clyde W. Hodge, Kristin K. Mehmert, Stephen P. Kelley, Thomas McMahon, Ashley Haywood, M. Foster Olive, Dan Wang, Ana Maria Sanchez-Perez & Robert O. Messing

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Correspondence to Clyde W. Hodge.

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Hodge, C., Mehmert, K., Kelley, S. et al. Supersensitivity to allosteric GABAA receptor modulators and alcohol in mice lacking PKCε. Nat Neurosci 2, 997–1002 (1999). https://doi.org/10.1038/14795

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