High-throughput caveolar proteomic signature profile for maternal binge alcohol consumption

Abstract

Currently, no single marker is sensitive and specific enough to be considered a reliable biomarker for prenatal alcohol exposure. To identify a proteomic signature profile for maternal alcohol consumption, we performed high throughput proteomics on maternal endothelial caveolae exposed to moderate binge-like alcohol conditions. In these specialized lipid ordered microdomains which contain a rich assembly of proteins, we demonstrate that moderate binge-like alcohol resulted in a distinctive maternal caveolar proteomic signature with important proteins being dramatically decreased/knocked out in the alcoholic profile. These proteins span from histones and basic structural proteins like tubulin α to proteins involved in trafficking, deubiquitination, cell signaling, and cell-cell adhesion. The profile also suggests an important role for the mother and the utero-placental compartment in the pathogenesis of Fetal Alcohol Spectrum Disorders (FASD). These data demonstrate that the caveolar proteomic signature created by alcohol shows a promising direction for early detection of FASD.

Alcohol and Maternal Uterine Vascular Adaptations During Pregnancy-Part I: Effects of Chronic In Vitro Binge-Like Alcohol on Uterine Endothelial Nitric Oxide System and Function

“…In this study, we observed that total cav-1 was significantly decreased in the LD alcohol group and was barely detected in the HD group. Our data support earlier studies where alcohol has been demonstrated to disrupt the caveolae by caveolar cholesterol/lipid depletion and disruption of caveolar assembly of proteins (Mao et al, 2009; Ramadoss et al, 2010; Ronis et al, 2007). Collectively, these data suggest that chronic binge-like alcohol has deleterious effects on the caveolar lipid rafts.…”

“…In addition, to mimic clinically relevant abusive patterns of drinking in women of child-bearing age and those who are admitted to emergency wards (Church and Gerkin, 1988; Hammond et al, 1973; Urso et al, 1981; Wells and Barnhill, 1996), we utilized a higher dose (HD, 600 mg/dl) group. Cells were exposed to alcohol in sealed, humidified chambers equilibrated with aqueous alcohol for 3 h on 3 consecutive days (Eysseric et al, 1997; Ramadoss et al, 2010; Ramadoss et al, 2007a; Ramadoss et al, 2007b), a pattern common among drinking women of child bearing age (Caetano et al, 2006; Gladstone et al, 1996; Maier and West, 2001). Alcohol concentrations were validated using an enzymatic assay kit (Quantichrom® ethanol assay kit, BioAssay Systems, Hayward, CA; data not shown).…”

Endothelial caveolar subcellular domain regulation of endothelial nitric oxide synthase

“…28 This chaperon protein HSP90 is also found to co-immunoprecipitate with eNOS under basal unstimulated condition in bovine aortic endothelial cells 28 and within the caveolae microdomain in ovine uterine artery endothelial cells. 29 HSP90 and eNOS interaction was observed to increase after receptor-mediated activation of eNOS; which was confirmed by the use of HSP90 specific inhibitor, Geldanamycin that leads to a reduced NO production. 28 …”

Effects of l-glutamine supplementation on maternal and fetal hemodynamics in gestating ewes exposed to alcohol

“…A study in pregnant C57BL/6 J mice demonstrated that NO modulation of the systemic mesenteric artery vascular response was hampered due to alcohol exposure (Cook et al 2001). In vitro studies have shown that binge-like alcohol exposure impairs eNOS activation and reduces mRNA levels for angiogenic genes and related proteome in endothelial cells of the ovine uterine artery, hence blunting the uterine vascular adaptations, including vasodilatory and angiogenic pathways (Ramadoss et al 2010, 2011; Ramadoss and Magness 2011, 2012a, b, d). Finally, these findings have significant implications on fetal growth and development.…”