Chronic exposure to alcohol results in neuroadaptive phenomena, including tolerance, sensitization, dependence, withdrawal, loss of control of drinking, and relapse that contribute to the development of excessive alcohol consumption. The goal of the INIA (Integrative Neuroscience Initiative on Alcoholism) Consortium is to identify the molecular, cellular, and behavioral neuroadaptations that occur in the brain reward circuits associated with the extended amygdala and its connections. It is hypothesized that genetic differences and/or neuroadaptations in this circuitry are responsible for the individual differences in vulnerability to the excessive consumption of alcohol. We propose to use quantitative proteomics to dissect the molecular mechanisms contributing to the behavioral phenotype of differential and excessive drinking (both baseline drinking and withdrawal induced drinking) in two animal models: 1) Adenylyl Cyclase 7 (AC7) transgenic animals - changes in the copy number of the AC7 gene produces changes in drinking phenotype using the Withdrawal Induced Drinking (WID) paradigm and 2) High Alcohol Preference (HAP)/Low Alcohol Preference (LAP) animals - animals selectively bred for differences in free-choice alcohol consumption by the 2 Bottle-Choice (2BC) paradigm - the selected genotype (changes in multiple genes) contributes to differential baseline drinking. Our goals are 1) to develop and optimize proteomic methodology for the quantitative analysis of enriched brain fractions, 2) to identify global differences in baseline protein expression between selected lines of the two animal models [AC7 transgenic model (AC7 transgenic versus wildtype) and HAP/LAP selective breeding model (HAP versus LAP)], and 3) to globally compare longitudinal changes in protein expression between animals (AC7 transgenic versus wildtype and HAP versus LAP) at selected time points during WID-2BC to identify proteins that contribute to the differential and excessive drinking behaviors.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project--Cooperative Agreements (U01)
Project #
7U01AA016653-06
Application #
8231796
Study Section
Special Emphasis Panel (ZAA1-DD (70))
Program Officer
Reilly, Matthew
Project Start
2006-09-30
Project End
2011-08-31
Budget Start
2011-03-10
Budget End
2011-08-31
Support Year
6
Fiscal Year
2010
Total Cost
$245,836
Indirect Cost
Name
University of Pittsburgh
Department
Physiology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Goulding, Scott P; Szumlinski, Karen K; Contet, Candice et al. (2017) A mass spectrometry-based proteomic analysis of Homer2-interacting proteins in the mouse brain. J Proteomics 166:127-137
Bateman, Nicholas W; Goulding, Scott P; Shulman, Nicholas J et al. (2014) Maximizing peptide identification events in proteomic workflows using data-dependent acquisition (DDA). Mol Cell Proteomics 13:329-38
Rogstad, Sarah M; Sorkina, Tatiana; Sorkin, Alexander et al. (2013) Improved precision of proteomic measurements in immunoprecipitation based purifications using relative quantitation. Anal Chem 85:4301-6
Goulding, Scott P; Maccoss, Michael J; Wu, Christine C (2013) Label-free differential analysis of murine postsynaptic densities. Methods Mol Biol 1002:295-309
Farias, Santiago E; Kline, Kelli G; Klepacki, Jacek et al. (2010) Quantitative improvements in peptide recovery at elevated chromatographic temperatures from microcapillary liquid chromatography-mass spectrometry analyses of brain using selected reaction monitoring. Anal Chem 82:3435-40
Kline, Kelli G; Frewen, Barbara; Bristow, Michael R et al. (2008) High quality catalog of proteotypic peptides from human heart. J Proteome Res 7:5055-61
Blackler, Adele R; Speers, Anna E; Wu, Christine C (2008) Chromatographic benefits of elevated temperature for the proteomic analysis of membrane proteins. Proteomics 8:3956-64
Grant, Kathleen J; Wu, Christine C (2007) Advances in neuromembrane proteomics: efforts towards a comprehensive analysis of membrane proteins in the brain. Brief Funct Genomic Proteomic 6:59-69