Effects of gonadotrophins and insulin on glucose uptake in the porcine cumulus–oocyte complex during IVM
Gabriel Martín Alvarez
A D E , María Josefina Barrios Expósito A , Evelin Elia B , Dante Paz B C , Sergio Morado A and Pablo Daniel Cetica A D
+ Author Affiliations
- Author Affiliations
A Area of Biochemistry, Institute of Research and Technology on Animal Reproduction, School of Veterinary Sciences, University of Buenos Aires, Chorroarín 280, CP 1427, Buenos Aires, Argentina.
B Developmental Biology Laboratory, Area of Biodiversity and Experimental Biology, Institute of Physiology, Molecular Biology and Neurosciences, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), School of Exact and Natural Sciences, University of Buenos Aires, Pabellon INFIBYNE, ingreso costanera norte, Ciudad Universitaria, CP 1428, Buenos Aires, Argentina.
C Biodiversity and Experimental Biology Department, School of Exact and Natural Sciences, University of Buenos Aires, Pabellon INFIBYNE, ingreso costanera norte, Ciudad Universitaria, CP 1428, Buenos Aires, Argentina.
D Institute of Research on Animal Production, Consejo Nacional de Investigaciones Científicas y Técnicas, Chorroarín 280, CP 1427, Buenos Aires, Argentina.
E Corresponding author. Email: galvarez@fvet.uba.ar
Reproduction, Fertility and Development 31(8) 1353-1359 https://doi.org/10.1071/RD18321
Submitted: 14 August 2018 Accepted: 7 February 2019 Published: 8 April 2019
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−1 LH + 10 IU mL−1 FSH) and 0.4 μU mL−1 insulin, during 44 h of IVM, on glucose transport and consumption, as well as on nuclear maturation and sperm penetration. We evaluated the effects of gonadotrophins 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 insulin alone 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.
Additional keywords: FSH, LH.
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