Studies with protein kinase C epsilon (PKC Epsilon) null mice indicate that PKC Epsilon regulates anxiety and alcohol consumption, dependence and reward. This appears to be related to actions at GABAA receptors since these mice are supersensitive to allosteric GABAA receptor agonists, including ethanol, in vivo and in vitro. The overall goal of this project is to understand how PKC Epsilon regulates GABAA receptor function. Studies will use immunohistochemistry and receptor binding autoradiography to compare the distribution of specific GABAA receptor subunits in wild type and PKC Epsilon null mice: Autoradiography will also be used to determine the affinity and density of binding sites for ligands that bind to benzodiazepine sites on specific receptor subpopulations. Western analysis will be used to measure the abundance of receptor subunits in limbic brain regions that express PKC Epsilon. L(tk-) fibroblast cell lines expressing different types of receptors will be examined to determine if PKC Epsilon regulation of allosteric sensitivity is specific for a particular subunit combination. Finally a PKC Epsilon mutant that can utilize novel ATP analogs as phosphate donors will be used to identify immediate substrates of PKC Epsilon in L(tk-) cells and in synaptoneurosomes, using immunoprecipitations with antibodies to GABAA receptor subunits and associated proteins, and 2D gel electrophoresis and mass spectrometry for unknown substrates. The hope is to define a PKC Epsilon pathway to further our understanding of GABAA receptor function as it relates to alcoholism.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
1R01AA013588-01
Application #
6460115
Study Section
Special Emphasis Panel (ZRG1-IFCN-1 (02))
Program Officer
Sorensen, Roger
Project Start
2002-05-01
Project End
2007-04-30
Budget Start
2002-05-01
Budget End
2003-04-30
Support Year
1
Fiscal Year
2002
Total Cost
$401,750
Indirect Cost
Name
Ernest Gallo Clinic and Research Center
Department
Type
DUNS #
173995366
City
Emeryville
State
CA
Country
United States
Zip Code
94608
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Maiya, Rajani; McMahon, Thomas; Wang, Dan et al. (2016) Selective chemical genetic inhibition of protein kinase C epsilon reduces ethanol consumption in mice. Neuropharmacology 107:40-48
Blasio, Angelo; Messing, Robert O (2016) Binge Drinking With Protein Kinase C Epsilon: A Role for Mammalian Target of Rapamycin Complex 2? Biol Psychiatry 79:425-6
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Lee, A M; Wu, D-F; Dadgar, J et al. (2015) PKC? phosphorylates ?4?2 nicotinic ACh receptors and promotes recovery from desensitization. Br J Pharmacol 172:4430-41
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Schuster, Daniel J; Kitto, Kelley F; Overland, Aaron C et al. (2013) Protein kinase C? is required for spinal analgesic synergy between delta opioid and alpha-2A adrenergic receptor agonist pairs. J Neurosci 33:13538-46

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