2-D DIGE uterine endothelial proteomic profile for maternal chronic binge-like alcohol exposure.

Little is known about alcohol effects on the utero-placental compartment during pregnancy. For the first time, we utilized 2-D DIGE quantitative proteomics to evaluate the role of the uterus in Fetal Alcohol Spectrum Disorders (FASD) pathogenesis. Uterine artery endothelial cells were isolated from pregnant ewes, FAC sorted, validated, and maintained in culture. To mimic maternal binge drinking patterns, cells were cultured in the absence or presence of alcohol (300 mg/dl) in a compensating sealed humidified chamber system equilibrated with aqueous alcohol for 3 h on 3 consecutive days for two weeks. CyDye switch combined with 2-D DIGE followed by MALDI-TOF and tandem MS/MS were utilized. Validation was performed using Western immunoblot analysis. Chronic binge-like alcohol significantly (P<0.05) decreased 30 proteins and increased 19 others. Gene-enrichment and functional annotation cluster analysis revealed significant enrichment (P<0.05) in three categories: glutathione S transferase, thioredoxin, and vesicle transport-related. Furthermore, alcohol differentially altered proteins with certain isoforms being downregulated while others were upregulated. In summary, binge alcohol has specific effects on the maternal uterine proteome, especially those related to oxidative stress. The current study also demonstrates a great need to utilize proteomic approaches for diagnostic, mechanistic and therapeutic aspects of FASD.