PATERNAL SHORT-TERM ALCOHOL CONSUMPTION ALTERS MOUSE EMBRYO-TROPHOBLAST DIFFERENTIATION DURING PERI-IMPLANTATION IN VITRO VIA PROGRAMMING EMBRYO DEATH

SALAMONE G; CALVO JC; FUENTES F.; CEBRAL E; GOTFRYD L; ERLEJMAN A; FONTANA V

Tokio

Congreso; International Federation of Placenta Associations (IFPA); 2018

Aim: To evaluate the effect of paternal alcoholconsumption on peri-implantation embryo development, focusing on trophoblastand inner cell mass differentiation and apoptosis. Methods: CF-1 malemice were exposed (treated group, T) or not (control group, C) to 15% (v/v)ethanol in drinking water ad libitum for15 days, followed by mating with non-treated CF-1 females (1:1). Pregnant females(positive vaginal plug) were sacrificed at day 2 of gestation to obtain 2-cellembryos which were cultured in vitro for 7 days. Embryo compaction, blastocoelexpansion and hatching were assessed during days 1-3 of culture(preimplantation); embryo-trophoblast growth and differentiation were analyzedduring days 4-7 of culture (in vitro implantation).At culture day 7, fixed embryos (4% paraformaldehyde) were classified as type A(ICM: protruding aggregates of compact cells; TB (trophoblasts): symmetric monolayerof flat and elongated cells) or type B (ICM: disaggregated, few scattered or nocells; TB: asymmetric trophoblast outgrowth). Embryo apoptosis was determinedby annexin V-FITC and RT-PCR. Results: Male alcohol consumption for 15days delayed embryo differentiation by deregulation of peri-implantation eventsand alteration of embryo morphogenesis. Distribution of A and B type embryoswas significantly different (Fisher´s exact test) between C and T groups. ICM:53% A in C vs 10% A in T (p