MICE PATERNAL ALCOHOL EXPOSURE AFFECTS EPIGENETICS MARKS ON SPERM ALTERING THE TESTIS STRUCTURE FROM ITS OFFSPRING

MAITE CAMBIASSO; LUCILA GOTFRYD; MARCELO STINSON; JUAN CARLOS CALVO; MARINA ROMANATO; VANINA FONTANA

Córdoba

Congreso; IXINTERNATIONAL MEETINGof the Latin American Society for Biomedical Research on Alcoholism (LASBRA); 2019

Previously, we observed that male alcoholconsumption increased the sperm rate of decondensation, delayedembryo differentiation by deregulating peri-implantation events andaltering embryo trophoblast and inner cell mass morphology invitro. This study evaluates theeffect of paternal alcohol consumption on spermatozoa and testis andits effect on male offspring. CF-1 male mice were exposed (treatedgroup, T) or not (control group, C) to 15% (v/v) ethanol in drinkingwater ad libitumfor 15 days. Sperm from epididymal cauda were obtained by swim-out todetermine sperm oxidative stress with the CellROXGreen Flow Cytometry Assay Kitand epigenetic marks of histone for immunocytochemistry. Testicleswere weighted and the DNA fragmentation was analyzed by TUNEL assayon both groups. Males from control and treated groups were mated withnon-treated CF-1 female mice in a ratio 1:1. Sperm from cauda ofadult male mice of the offspring (F1) were obtained by swim-out todetermine sperm parameters and head decondensation. Testicles of F1mice were weighted and analyzed histologically. Male alcoholconsumption did not alter testicular weight but increased spermoxidative stress (fluorescence intensity: 2,2±0,2 n=5 C vs 3,0±0,2n=4 T, p<0,01) On the other side, we observed a significantdecrease of epigenetic marks of histone H3K4me3 in sperm from T groupcompared to C group (positive mark: 7,0±3,7 n=8 T vs 27,1±8,2 n=11C, p<0,05). Besides, we detected an increment of TUNEL labeling ongerminal line from testicles of treated groups vs. control groups(56%±6% T vs 21%±4% C, p<0,01, n=2).When we analyzed F1 mice wecould detect differences in the testicles weight (0,099±0,004 g n=11T vs 0,118±0,006 g n=11 C, p<0,05) and their germinal linethickness from F1 male mice of treated group (275±3 µm, n=7 C vs249±8 µm, n=5 T, p<0,01) being both significantly minor thatthose in control group. However,there were not differences insperm concentration, motilityand head decondensation between both groups of F1 male mice. Takingtogether, these results suggest that short-term paternal alcoholconsumption impairs epididymal sperm quality altering the malereproductive biology and inheriting reproductive defects to theirmale offspring.