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

LUCILA GOTFRYD; GABRIELA SALAMONE; FERNANDO FUENTES; ALEJANDRA ERLEJMAN; JUAN CARLOS CALVO; ELISA CEBRAL; VANINA FONTANA

Tokyo

Congreso; International Federation of Placenta Associations Meeting; 2018

Aim: To evaluate the effect of paternal alcohol consumption on peri-implantation embryo development, focusing on trophoblast and inner cell mass differentiation and apoptosis. Methods: CF-1 male mice were exposed (treated group, T) or not (control group, C) to 15% (v/v) ethanol in drinking water ad libitum for 15 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-cell embryos which were cultured in vitro for 7 days. Embryo compaction, blastocoel expansion and hatching were assessed during days 1-3 of culture (preimplantation); embryo-trophoblast growth and differentiation were analyzed during 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 monolayer of flat and elongated cells) or type B (ICM: disaggregated, few scattered or no cells; TB: asymmetric trophoblast outgrowth). Embryo apoptosis was determined by annexin V-FITC and RT-PCR. Results: Male alcohol consumption for 15 days delayed embryo differentiation by deregulation of peri-implantation events and alteration of embryo morphogenesis. Distribution of A and B type embryos was significantly different (Fisher´s exact test) between C and T groups. ICM: 53% A in C vs 10% A in T (p<0.001). TB: 60% A in C vs 26% A in T (p<0.01). After 7 days of culture, apoptotic cells were detected in embryos of T vs C group. Conclusion: Short-term paternal alcohol consumption impairs mouse embryo peri-implantation by affecting ICM and TB differentiation and programming them to death. Considering that ICM participates in embryo formation and TB plays a relevant role in placental development, these effects might be crucial for further embryo survival.