Effects of gonadotrophins and insulin on glucose uptake in the porcine cumulus–oocyte complex during IVM

Abstract

The combination of gonadotrophins (LH and FSH) and insulin is frequently used in porcine oocyte IVM, but the individual effects of gonadotrophins and insulin have not been completely studied. The aim of this study was to investigate the mechanisms involved in glucose metabolism in the swine cumulus?oocyte complex (COC), analysing the effects of gonadotrophins (10 IU/mL LH + 10 IU/mL FSH) and 0.4 uU/mL insulin, during 44 h of IVM, on glucose transport and consumption, as well as on nuclear maturation and sperm penetration. We evaluated the effects ofgonadotrophins and insulin separately or in combination on glucose consumption, membrane permeability to the glucose fluorescent analogue 6-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-6-deoxyglucose (6-NBDG), the presence of GLUT-4 and oocyte maturation rates, after 44 h of IVM. Nuclear maturation percentages increased significantly following the addition of gonadotrophins alone or in combination with insulin to the culture medium (P < 0.0001), whereas insulinalone had no effect. A significant increase was observed in sperm penetration of COCs matured with insulin, gonadotrophins or their combination (P , 0.0001). However, only gonadotrophins significantly increased glucose uptake (P < 0.0001). Although gonadotrophins and insulin increased GLUT-4 expression, neither modified 6-NBDG incorporation. In conclusion, gonadotrophins and insulin had different effects during IVM; although gonadotrophins increased maturation rates and glucose consumption, they had no effect on glucose transport, and insulin improved sperm penetration without affecting the parameters related to glucose utilisation. Therefore, glucose metabolism is likely to be primarily regulated by its consumption in metabolic pathways rather than by changes in membrane permeability.