The overall goal is to identify the neurochemical mechanisms responsible for ethanol and benzodiazepine intoxication, tolerance and dependence. We postulate that the GABA-activated chloride channel is an important site of action for these drugs and the proposed research will focus exclusively on this receptor-channel complex. Of particular importance is the use of molecular biological techniques to define genetic differences in alcohol and benzodiazepine sensitivity at the level of the individual genes coding for the GABA-activated chloride channel complex. The proposal has 4 aims: 1. GABA-activated chloride channels are modulated by endogenous regulators such as calcium, cAMP and protein kinase C. The possibility that these regulators modulate the acute actions of ethanol or benzodiazepines or are responsible for tolerance to these drugs will be studied. 2. The channel complex consists of alpha, beta and perhaps gamma subunits; these distinct subunits have been cloned and sequenced, but it is not known if they confer different ethanol or benzodiazepine sensitivity on the channel. We will answer this question by testing drug sensitivity after expression of the cloned subunits in oocytes and by creation of transgenic mice with over- or under-expression of genes coding for specific subunits. The transgenic mice will allow us to ascribe functional roles to the different subunits. 3. Brain mRNA from Long/Short-Sleep (LS/SS) and Diazepam Sensitive/Resistant (DS/DR) mice and RNA from cloned genes will be expressed in Xenopus oocytes to allow electrophysiological study of channel function. Cloned genes from drug- sensitive mice (LS,DS) will also be injected into embryos of drug- resistant mice (SS,DR) to produce transgenic mice with expression of genes from resistant and sensitive lines. 4. Genetic differences in behavioral sensitivity to ethanol or benzodiazepines may be due to differences in nucleotide sequences of the genes coding for the subunits of the GABA-activated chloride channel or sequences of genes coding for promoter regions. Use of LS/SS and DS/DR mice for gene cloning will identify sequence differences between the lines.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AA006399-13
Application #
2043455
Study Section
Special Emphasis Panel (NSS)
Project Start
1983-09-29
Project End
1999-08-31
Budget Start
1995-09-01
Budget End
1996-08-31
Support Year
13
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of Colorado Denver
Department
Pharmacology
Type
Schools of Medicine
DUNS #
065391526
City
Aurora
State
CO
Country
United States
Zip Code
80045
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Blednov, Yuri A; Black, Mendy; Benavidez, Jillian M et al. (2017) Sedative and Motor Incoordination Effects of Ethanol in Mice Lacking CD14, TLR2, TLR4, or MyD88. Alcohol Clin Exp Res 41:531-540
Borghese, Cecilia M; Ruiz, Carlos I; Lee, Ui S et al. (2016) Identification of an Inhibitory Alcohol Binding Site in GABAA ?1 Receptors. ACS Chem Neurosci 7:100-8
Blednov, Yuri A; Black, Mendy; Benavidez, Jillian M et al. (2016) PPAR Agonists: II. Fenofibrate and Tesaglitazar Alter Behaviors Related to Voluntary Alcohol Consumption. Alcohol Clin Exp Res 40:563-71
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Blednov, Yuri A; Black, Mendy; Benavidez, Jillian M et al. (2016) PPAR Agonists: I. Role of Receptor Subunits in Alcohol Consumption in Male and Female Mice. Alcohol Clin Exp Res 40:553-62
Blednov, Yuri A; Benavidez, Jillian M; Black, Mendy et al. (2015) Peroxisome proliferator-activated receptors ? and ? are linked with alcohol consumption in mice and withdrawal and dependence in humans. Alcohol Clin Exp Res 39:136-45
Borghese, Cecilia M (2015) The molecular pharmacology of volatile anesthetics. Int Anesthesiol Clin 53:28-39

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