Implantational in vitro development of trophoblast after perigestational alcohol consumption, in CD-1 mouse model

PEREZ TITO L; CEBRAL E; BEVILACQUA E

San Pablo

Congreso; XV Congresso da Sociedade Brasileira de Biologia Celular (SBBC; 2010

SBBC

Periconceptional alcohol consumption in CF-1 mice alters preimplantational embryo differentiation. We studied the effects of maternal alcohol ingestion by CD-1 mice on in-vivo (day 5 of gestation) and in-vitro trophoblastic growth, proliferation, migration and differentiation. Adult females were exposed to 10% ethanol in drinking water for 15 days previous to and during 5 days of gestation (TF) (Control group (CF) with water). Recovered hatched blastocysts were cultured and at 0, 24, 48 and 72 h were analyzed for trophoblastic expansion (TE) areas and embryo morphology. Toluidine blue stain or immunocitochemistry for Ki67 was performed. Embryos were classified as smaller and bigger ones according to intervals of TE areas. The nuclear number (Nr) and trophoblastic (T) differentiation was evaluated. At day 5, TF presented diminished Nr of blastocyts/female (p<0.05) and elevated abnormal embryo % (p<0.05) vs CF. While smaller or bigger embryo % was invariable in CF through 72 h-culture, the TF-bigger embryos % increased at 48 h (p<0.05 vs 48 h-CF, p<0.001 vs 0 and 24 h-TF). The total trophoblastic nuclei Nr resulted decreased in TF (p<0.05 vs CF) although T-differentiation (negative Ki67-immunostain) did not change. While CF-smaller embryo areas increased at 48 h-culture (p<0.05 vs 0 h), the TF-smaller ones increased at 24 h (p<0.001). TF-smaller embryo expansion increased 25% respect to CF-embryos in 0-24 and 24-48 h-culture, but TF-bigger embryo areas reduced 14% of CF at 0-24 h-culture. These results suggested implantating embryo losses, morphologic and proliferation alterations and deregulation of trophoblastic expansion/migration, following maternal alcohol ingestion.