EFFECT OF TROLOX ON PLASMA MEMBRANE INTEGRITY AND MITOCHONDRIAL FUNCTION IN REFRIGERATED PORCINE SEMEN

NICOLÁS GARCÍA; MARIANA CÓRDOBA; PABLO CETICA; SERGIO MORADO

Buenos Aires

Jornada; XX JORNADAS ANUALES DE LA SOCIEDAD ARGENTINA DE BIOLOGÍA (SAB), XVII JORNADAS DE LA SOCIEDAD URUGUAYA DE BIOCIENCIAS (SUB)-Segundas Jornadas Rioplatenses de Biología; 2018

Gamete refrigeration is a commonly used biotechnology for porcine production. Porcine sperm have a high proportion of unsaturated lipids in their plasma membrane making them more susceptible to damages caused by temperature descent. Trolox (6-hydroxi-2,5,7,8-tetrametilchroman-2-ácido carboxilico) is a vitamin E analogue used as antioxidant, which proved to improve sperm quality post-freezing in several species. In the porcine, sperm have a high oxidative metabolism and energy is generated mainly in the mitochondria. The aim of our work was to study the changes in functional parameters in porcine sperm refrigerated with or without Trolox. Sperm capacitation was analyzed using the epifluorescent chlorotetracycline technique (CTC), plasma membrane integrity by hypoosmotic test (HOS-test), sperm vitality using Trypan Blue, acrosome integrity by a differential interferential microscope (DIC), mitochondrial membrane potential by 5,5´, 6,6´- tetracloro-1,1´, 3,3´yoduro de tetra-etil-benzimidazolil-carbocianina (JC-1) stain and in vitro fertilization by the observation of pronuclei formation. Samples were suspended in a commercial diluent in a 1:10 proportion and treated with or without Trolox at 17 ºC. Data were analyzed using ANOVA and Tukey test (P0.05). Sperm vitality in refrigerated samples was improved by the addition of Trolox (P0.05). Regarding mitochondrial membrane potential, the lowest level was observed in samples refrigerated without the antioxidant (P