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Neuroscience Graduate Program at UCSF
Signaling Pathways in Alcoholism and Neurologic Disease
My laboratory is interested in the molecular basis of behavior, especially as it relates to substance abuse and co-morbid neurological and psychiatric conditions, such as anxiety and pain. Studies in cell culture and in model systems such as Drosophila melanogaster have identified signaling proteins important for acute and chronic responses to ethanol and other drugs of abuse. The lab uses gene targeting and RNA interference in rodents to understand the role of these proteins in behavior and to determine if they are targets for development of new therapeutic agents to treat addiction. In addition, we have pursued studies with some of these proteins in stroke and Alzheimer’s disease.
Protein kinase C (PKC)e: We found that PKCe null mice show: (1) supersensitivity to drugs that activate GABAA receptors; (2) decreased acute tolerance to ethanol and decreased ethanol self-administration; (3) decreased anxiety and down-regulation of the HPA axis; and (4) decreased hyperalgesia associated with inflammation or alcoholic polyneuropathy. We are now investigating molecular mechanisms by which PKCe modulates GABAA receptors and are mapping brain regions where PKCe regulates anxiety and alcohol responses. We have screened compound libraries and identified a family of lead compounds as selective PKCe inhibitors.
PKCd: We have generated PKCd null mice and found that they are very resistant to intoxication by ethanol. This is associated with reduced responses to other drugs that act at extra-synaptic GABAA receptors. We are investigating whether PKCd regulates these GABAA receptors directly or indirectly through GABA transporters. We also found that these mice are substantially resistant to brain injury resulting from focal ischemia and reperfusion. We are now examining PKCd signaling pathways that mediate this resistance.
PKC substrates: The catalytic domains of the PKC family are very similar and in vitro studies have failed to identify PKC isozyme-specific substrates. In collaboration with Kevan Shokat at UCSF, we have generated PKCe and PKCd variants with mutations in their ATP binding domains to confer sensitivity to inhibitors that are inactive at normal kinases. We are expressing these mutant kinases in cells and generating knock-in mice to identify PKC isozyme-specific substrates.
N-type calcium channels:We previously found that ethanol inhibits N-type calcium channels and chronic ethanol exposure increases N channel density in the brain. We recently found that mice lacking N-type channels show reduced sensitivity to the hypnotic effect of ethanol, altered ethanol reward and reduced ethanol preference compared with wild type mice. We are currently investigating whether novel small molecule N-type channel antagonists decrease drinking in rodents.
Diacylglycerol kinases: Drosophila with mutations in the gene rgdA show altered activity in response to ethanol. This gene encodes for a protein that is a member of the family of diacylglycerol kinases and is homologous to mammalian DGKi and DGKz, that are both expressed widely in brain. DGKs metabolize diacylglycerol and antagonize effects of PKC isozymes if they lie within the same subcellular compartment. We are investigating behavioral responses to alcohol in mice lacking each of these DGKs to determine if they should be considered as new targets for treatment of substance abuse.
Lab Personnel (Current)
Lisa Wallace
Postdoctoral Fellow
PhD, Virginia Commonwealth Univ.
PKC regulation of cannabinoid signaling
Zhan-heng Qi
Postdoctoral Fellow
PhD, Beijing Univ.
PKC regulation of GABA-A receptor function; PKC substrates in nociception
Lab personnel (Former)
Heidi Lesscher
Postdoctoral Fellow
2004-2006
PKC epsilon, CRF and anxiety
PhD, U. of Utrecht, Netherlands
Postdoc, Rudolf Magnus Inst. For Neuroscience, U. of Utrecht
Miho Oyasu
Postdoctoral Fellow
2003-2006
PKC activation by ethanol
PhD, Kobe U., Japan
Scientist, A-Cube, Burlingame, CA
Phil Newton
Postdoctoral Fellow
2001-2005
N channels and alcohol; PKC and memory
PhD, Univ. of Birmingham, UK
Asst Prof, UCSF; Assoc Invest, Gallo
Wen-Hai Chou
Postdoctoral Fellow; Scientist
2001-2005
PKC regulation of GABA-A receptor trafficking; PKC delta and stroke
PhD, Univ. of TX, San Antonio
Asst Prof, UCSF; Assoc Invest, Gallo
Maengseok Song
Postdoctoral Fellow; Scientist
2001-2005
Regulation of GABA-A receptors by PKC epsilon
PhD, Korean Advanced Inst. For Science and Technology
Technical Support, Applied Biosystems
Doo-Sup Choi
Postdoctoral Fellow; Scientist
1999-2005
Alcohol responses in ENT1 and PKCdelta null mice; PKCepsilon and A-beta degradation
PhD, Univ. of Strasbourg, France
Asst. Prof, Mayo Medical School, Rochester, MN
Helen Walter
Postdoctoral Fellow
1998-2002
Ethanol regulation of calcium channels
PhD, U. of Leeds, UK
Teacher, Peralta College, Alameda, CA
Annick Martin
Postdoctoral Fellow
1997-2001
PKC epsilon in pain responses
PhD, U. Pierre et Marie Curie, France
Project Leader, Assoc. Francise Contre les Myopathies, France
Effie Lin
Postdoctoral Fellow
1996-1998
Generation of PKC epsilon null mice
PhD, Stanford University
Scientist, Gentastic Corp.
Ed Gerstin
Postdoctoral Fellow
1995-1997
Regulation of L-type calcium channels by ethanol
PhD, Univ. Calif., Irvine
Scientist, Incyte Corp.
Michele Solem
Postdoctoral Fellow
1993-1995
L type calcium channels and neural differentiation
PhD, Oregon State University
Assoc. Professor Thomas Jefferson Univ.
Bhupinder Hundle
Postdoctoral Fellow
1992-1997
PKC epsilon and neural differentiation
PhD, Univ. Calif.,
Berkeley
European Sales Director, Amaxa, UK
Reina Roivainen
Postdoctoral Fellow
1991-1993
Ethanol-induced neural plasticity
MD, PhD, U. Tampere, Finland
Neurologist, Helsinki U. Hospital
Link to Publication via PubMed
Khasar SG, Lin Y-H, Martin A, Dadgar J, McMahon T, Hundle B, Aley KO, Isenberg W, Green PG, Hodge, CH, Levine JD, Messing RO. A novel nociceptive signaling pathway demonstrated in PKCε-mutant mice. Neuron 24:253-260, 1999.
Hodge, CW, Mehmert K, Kelley SP, McMahon T, Haywood A, Olive MF, Wang D, Sanchez-Perez AM, Messing RO. Supersensitivity to allosteric GABAA modulators and alcohol in mice lacking PKCε. Nature Neurosci. 2:997-1002, 1999.
McMahon T, Andersen R, Messing RO. PKCε mediates up-regulation of N-type calcium channels by ethanol. Mol. Pharmacol. 57:53-58, 2000.
Aley KO, Messing RO, Mochly-Rosen D, Levine JD. Chronic hypersensitivity for inflammatory nociceptor sensitization mediated by the ε isozyme of protein kinase. J. Neurosci. 20:4680-4685, 2000.
Walter H, McMahon T, Gerstin E, Dadgar J, Wang D, Messing RO. Ethanol regulates calcium channel subunits by protein kinase C δ-dependent and -independent mechanisms. J. Biol. Chem. 275: 25717-25722, 2000.
Dina OA, Barletta J, Chen XJ, Mutero A, Martin A, Messing RO and Levine JD. Key role for the ε isoform of protein kinase C in painful alcoholic neuropathy in the rat. J. Neurosci. 20:8614-8619, 2000.
Aley KO, Martin A, McMahon T, Levine JD, Messing RO. Nociceptor sensitization by extracellular signal-regulated kinases. J. Neurosci. 21:6933-6939, 2001.
Hodge CW, Raber J, Walter H, McMahon T, Sanchez-Perez AM, Olive MF, Mehmert K, Morrow AL, Messing RO. Decreased anxiety-like behavior, reduced stress hormones, and neurosteroid supersensitivity in mice lacking protein kinase C ε. J. Clin. Invest. 110:1003-1010, 2002.
Choi D-S, Wang D, Dadgar J, Chang WS, Messing RO. Conditional rescue of protein kinase C ε regulates ethanol preference and hypnotic sensitivity in adult mice. J. Neurosci. 22:9905-9911, 2002.
Chou W-H, Choi D-S, Zhang H, Mu D, McMahon T, Kharazia VN, Lowell CA, Ferriero DM, Messing RO. Neutrophil protein kinase C δ as a mediator of stroke-reperfusion injury. J. Clin. Invest. 114:49-56, 2004.
Choi D-S, Cascini M-G, Mailliard W, Young H, Paredes P, McMahon T, Diamond I, Bonci A, Messing RO. The equilibrative nucleoside transporter type 1 modulates ethanol intoxication and preference in mice. Nat. Neurosci. 7:855-861, 2004.
Newton PM, Orr CJ. Wallace MJ, Shin H-S, Messing RO. Deletion of N-type calcium channels blocks ethanol reward and reduces ethanol consumption in mice. J. Neurosci. 24:9862-9869, 2004.
Olive MF, McGeehan AJ, Kinder JR, McMahon T, Hodge CW, Janak PH, Messing RO. The mGluR5 antagonist 6-methyl-s-(phenylethynyl)pyridine decreases ethanol consumption via a protein kinase C ε-dependent mechanism. Mol. Pharmacol. 67:349-355, 2005.
Wallace MJ, Newton PM, Oyasu M, McMahon T, Chou W-H, Connolly J, Messing RO. Acute functional tolerance to ethanol mediated by protein kinase Cε. Neuropsychopharmacology, March 15 [Epub ahead of print], 2006.
Choi DS, Wang D, Yu G-Q, Zhu G, Kharazia VN, Paredes P, Chang WS, Deitchman JK, Mucke L, Messing RO. PKCε increases endothelin converting enzyme activity and reduces amyloid plaque pathology in transgenic mice. Proc. Nat. Acad. Sci. USA, 103:8215-8220, 2006.
Robert Messing, M.D.
Phone
510-985-3100
Office Location
Ernest Gallo Clinic & Research Center
5858 Horton Street
Suite 200
Emeryville, CA 94608
Other Websites
Ernest Gallo Clinic and Research Center