Effect of AMP on the oxidative metabolism of the bovine oocyte during in vitro maturation

CYNTHIA GUTNISKY; SERGIO MORADO; GABRIEL DALVIT; JEREMY THOMPSON; PABLO CETICA

Buenos Aires

Jornada; III Jornadas Internacionales del Instituto de Investigación y Tecnología en Reproducción Animal; 2012

Facultad de Ciencias Veterinarias, Universidad de Buenos Aires

The uptaken glucose by cumulus-oocyte complexes (COCs) during in vitro maturation might be involved in the regulation of oxidative metabolism. The aim of this work was to study the effect of the COC glycolytic pathway stimulation on the glucose uptake, lactate production and in vitro maturation of the oocyte. A further aim was to determine the effect of stimulation of COC glycolysis on oocyte oxidative and mitochondrial activities. COCs were matured individually in medium 199 + 5% FBS with or without gonadotrophic supplementation (control) and without hormonal supplementation but with AMP (0, 1, 10, 20 and 40 mM). Glucose uptake and lactate production from spent media was measured by spectrophotometric assays. The cytoplasmic maturation was evaluated by the ratio of cleaved embryos after in vitro fertilization. The oxidative and mitochondrial activities of the oocytes were measured at 4 different time points (0, 9, 15, 22h), oocytes were stained with the fluorescent probes Redox Sensor Red CC-1 and MitoTracker Green FM and observed by laser scanning confocal microscopy. The images were analysed using Photoshop 9.0 software. COCs matured with AMP had no effect on glucose uptake and lactate production. However, in the absence of gonadotrophins a dose dependent stimulatory effect was observed (p