Periconceptional Ethanol Consumption Alters Collagens and Metalloproteinase-9 Expression in CF-1 Murine Implantation Sites at Midgestation

COLL TAMARA ANAHÍ; FONTANA VANINA; FAVARO RODOLFO R; PÉREZ TITO LETICIA GABRIELA; IGLESIAS AYELÉN ALUMINÉ; CALVO JUAN CARLOS; TENORIO ZORN TELMA MARÍA; CEBRAL ELISA

Santiago, Chile

Congreso; IFPA Meeting 2010; 2010

International Federation of Placenta

Periconceptional alcohol ingestion causes organogenic embryo anomalies, growth restriction and early miscarriages. The MMP-9 digests types IV and V collagens, playing a role in remodeling the extracellular matrix (ECM), decidualization, angiogenesis and vascularization, trophoblastic migration and invasion of decidua. Objective: to evaluate if periconceptional ethanol ingestion alters distribution, localization of collagens types I, III, IV and V, and MMP-9 expression/activity in vascular mesometrial decidua (VMD) and trophoblastic (T) tissues in organogenic implantation site. Methods: adult CF-1 female mice were treated (TG) with 10% ethanol in drinking water for 17 days previous to and during gestation up to day 10 of pregnancy. Control group (CG) were administered with water and mated. Implantation sites were processed for picrosirius staining for fibrillar collagens, and  immunohistochemistry for I, III, IV, V collagens and MMP-9. Decidual tissues were dissected to perform Western blotting for MMP-9 and zymograms of conditioned media from D cultured for 24 h. Results: while a defined and continuous picrosirius stain in basal membranes of vessel endothelium in VMD-CG was observed, the VMD-TG picrosirius stain was undefined and discontinuous. Collagen III had similar immunostaining in VMD-CG and VMD-TG. The immunofluorescence pattern for IV and V collagens increased in VMD-TG vs VMD-CG. A slight MMP-9 reactivity was detected in VMD-CG and in T-CG, while the immunoreactivity was reduced in VMD-TG and T-TG. In D-TG, the MMP-9 expression after immunoblotting resulted significantly decreased as compared to the D-CG. Zymographic analysis for active MMP-9 in conditioned medium resulted diminished in D-TG vs D-CG. Discussion: periconceptional alcohol ingestion reduce MMP-9 expression and activity, suggesting a diminished collagens IV and V digestion, while picrosirius staining indicates structural alterations in fibrillar collagens. These effects may be related to early anomalies in decidual-placental development and the increased spontaneous miscarriage after maternal alcohol consumption.