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

CAMBIASSO MAITE YAEL; LUCILA GOTFRYD; STINSON MARCELO GABRIEL; CALVO, JUAN CARLOS

Cordoba

Congreso; IX INTERNATIONAL MEETING of the Latin American Society for Biomedical Research on Alcoholism (LASBRA); 2019

LASBRA

MICE PATERNAL ALCOHOL EXPOSURE AFFECTS EPIGENETICS MARKS ONSPERM ALTERING THE TESTIS STRUCTURE FROM ITS OFFSPRINGCambiasso, Maite1, Gotfryd, Lucila2 , Stinson, Marcelo Gabriel2 , Calvo, Juan Carlos1,2, Romanato, Marina1 , & Fontana, Vanina Andrea21Instituto de Biología y Medicina Experimental (IBYME-CONICET),Ciudad Autónoma de Buenos Aires, Argentina.2Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales(IQUIBICEN-CONICET). Universidad de Buenos Aires (UBA).Ciudad Autónoma de Buenos Aires, Argentina.v_fontana@yahoo.comPreviously, we observed that male alcohol consumption increased the sperm rate of decondensation, delayed embryo differentiation by deregulating peri-implantation events and altering embryo trophoblast and inner cell mass morphology in vitro. This study evaluates the effect of paternal alcohol consumption on spermatozoa and testis and its effect on male offspring. 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. Sperm from epididymal cauda were obtained by swim-out to determine sperm oxidative stress with the CellROX Green Flow Cytometry Assay Kit and epigenetic marks of histone for immunocytochemistry. Testicles were weighted and the DNA fragmentation was analyzed by TUNEL assay on both groups. Males from control and treated groups were mated with non-treated CF-1 female mice in a ratio 1:1. Sperm from cauda of adult male mice of the offspring (F1) were obtained by swim-out to determine sperm parameters and head decondensation. Testicles of F1 mice were weighted and analyzed histologically. Male alcohol consumption did not alter testicular weight but increased sperm oxidative stress (fluorescence intensity: 2.2 ± 0.2 n = 5 C vs 3.0 ± 0.2 n = 4 T, p < .01) On the other side, we observed a significant decrease of epigenetic marks of histone H3K4me3 in sperm from T group compared to C group (positive mark: 7.0 ± 3.7 n = 8 T vs 27.1 ± 8.2 n = 11 C, p < .05). Besides, we detected an increment of TUNEL labeling on germinal line from testicles of treated groups vs. control groups (56% ± 6% T vs 21% ± 4% C, p < .01, n = 2). When we analyzed F1 mice we could detect differences in the testicles weight (0.099 ± 0.004 g n = 11 T vs 0.118 ± 0.006 g n = 11 C, p < .05) and their germinal line thickness from F1 male mice of treated group (275 ± 3 µm, n = 7 C vs 249 ± 8 µm, n = 5 T, p < .01) being both significantly minor that those in control group. However, there were not differences in sperm concentration, motility and head decondensation between both groups of F1 male mice. Taking together, these results suggest that short-term paternal alcohol consumption impairs epididymal sperm quality altering the male reproductive biology and inheriting reproductive defects to their male offspring.Grants: Honorio Bigand Foundation