EFFECT OF ZN ON SPERM PHYSIOLOGY

FERNÁNDEZ BEATO, LUCIANA; CANE, FERNANDO; PISKULIC LAURA; MARINI PATRICIA; TEIJEIRO JUAN MANUEL

Congreso; Reunión Anual de Sociedades de Biociencias SAIC, SAI, SAFIS; 2020

Optimal preservation ofboar sperm for artificial insemination (AI) is at 16°C. At this temperature, neithercellular metabolism nor microbiological growth conditions can be effectivelyreduced in seminal doses. Under the hypothesis that Zn?s effect on spermphysiology could improve their preservation at lower temperatures, theobjective of this work was to evaluate Zn supplementation on vitality (eosin-exclusiontest), motility (Computer-Assisted Sperm Analysis, CASA) and reactive oxygenspecies concentration (ROS, luminol) in seminal doses stored at 4°C. Also,mitochondrial and DNA integrities were evaluated by developing a new techniquebased on sperm chromatin dispersion test and cytochrome c oxidase activity.Aliquots of seminal doses were supplemented with 0 (Control, C), 0.5, 1, 2 or 3mM ZnCl2 and kept at 4°C for three days. Vitality, motility and ROS concentrationdata were analysed by ANOVA or Kruskal-Wallis in the case the assumptions were notmet by ANOVA. Mitochondrial and DNA integrity data were analysed by linearmixed models for longitudinal data. The percentage of motile sperm was higherin doses supplemented with 1 and 2 mM ZnCl2 by day 2 (63.25aC vs 68.5a 0.5 mM vs 73.75b 1 mM vs 71.5b 2 mMvs 65.5c 3 mM de ZnCl2, p=0.082) while, by day 3 alldoses supplemented with Zn showed higher percentage of motile sperm than thecontrol (47.75a C vs 61b 0.5 mM vs 65.75b 1 mMvs 60.50b 2 mM vs 61.50b 3 mM ZnCl2, p=0.0184).CASA analysis demonstrated differences in the linear pattern of movement andpercentage of rapid sperm. Mitochondrial integrity was affected only by thetime of storage while vitality, ROS concentration and DNA integrity did notshow changes neither by treatment nor time of the experiment. The resultsindicate that Zn preserves motility of sperm storage at low temperatureallowing its use for AI.