Effects of ethanol drinking on protein expression in rat amygdala studied by quantitative proteomics using dimethyl labeling

Wednesday, November 06, 2013 — Poster Session I
4:00 p.m. – 6:00 p.m.	FAES Academic Center (Upper-Level Terrace)	NIAAA	MOLBIO-10

Authors

Z-M Ding
W.J McBride
H-Y Kim

Abstract

Alcoholism is caused by ethanol-induced neuronal alterations within the brain’s reward neurocircuitry. The amygdala is thought to be one of primary neural structures associated with alcohol addiction. However, detailed biochemical mechanisms governing the alcohol effect at molecular and cellular levels are not fully understood. The objective of the present study was to determine the effect of ethanol drinking on protein expression level in rat amygdala. Rats were given one of three ethanol access conditions for four weeks: continuous ethanol (CE), multiple scheduled (MSA), or EtOH-naïve (water). Proteins were extracted from dissected amgydala and subjected to SDS-PAGE/in-gel trypic digestion. Light, intermediate or heavy dimethyl labeling was performed on peptides from MSA, CE or water groups, respectively. Differentially labeled samples from each fraction were mixed, desalted and analyzed by nano-LC-ESI-MS/MS. Protein identification and quantitation were performed using Mascot Distiller software. Base on biological triplicates, our results indicated that 70 proteins were either up-or down- regulated in ethanol drinking groups compared to water control group. Interestingly, MSA and CE differentially altered the expression levels for 35 and 12 proteins respectively. These results will be validated by western blotting or RT-PCR and/or subjected to network-pathway analysis