title: Pentose phosphate pathway activity: effect on in vitro maturation and oxidative status of bovine oocytes.
abstract: The relationship between pentose phosphate pathway (PPP) activity in cumulus-oocyte complexes (COCs) and oxidative and mitochondrial activity in bovine oocytes was evaluated with the aim of analysing the impact of two inhibitors (NADPH and 6-aminonicotinamide (6-AN)) and a stimulator (NADP) of the key enzymes of the PPP on the maturation rate, oxidative and mitochondrial activity and the mitochondrial distribution in oocytes. The proportion of COCs with measurable PPP activity (assessed using brilliant cresyl blue staining), glucose uptake, lactate production and meiotic maturation rate diminished when 6-AN (0.1, 1, 5 and 10mM for 22h) was added to the maturation medium (P<0.05). The addition of NADPH did not modify glucose uptake or lactate production, but reduced PPP activity in COCs and meiotic maturation rates (P<0.05). The presence of NADP (0.0125, 0.125, 1.25 and 12.5mM for 22h of culture) in the maturation medium had no effect on PPP activity in COCs, glucose uptake, lactate production and meiotic maturation rate. However, in the absence of gonadotropin supplementation, NADP stimulated both glucose uptake and lactate production at 12.5mM (the highest concentration tested; P<0.05). NADP did not modify cleavage rate, but decreased blastocyst production (P<0.05). During IVM, oocyte oxidative and mitochondrial activity was observed to increase at 15 and 22h maturation, which was also related to progressive mitochondrial migration. Inhibiting the PPP with 6-AN or NADPH led to reduced oxidative and mitochondrial activity compared with the respective control groups and inhibition of mitochondrial migration (P<0.05). Stimulation of the PPP with NADP increased oxidative and mitochondrial activity at 9h maturation (P<0.05) and delayed mitochondrial migration. The present study shows the significance of altering PPP activity during bovine oocyte IVM, revealing that there is a link between the activity of the PPP and the oxidative status of the oocyte.
authors.affiliation: Area of Biochemistry, Institute of Research and Technology on Animal Reproduction, School of Veterinary Sciences, University of Buenos Aires, Chorroarín 280, Buenos Aires C1427CWO, Argentina.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Gutnisky
authors.affiliation: Area of Biochemistry, Institute of Research and Technology on Animal Reproduction, School of Veterinary Sciences, University of Buenos Aires, Chorroarín 280, Buenos Aires C1427CWO, Argentina.
authors.forename: Gabriel C
authors.initials: GC
authors.lastname: Dalvit
authors.affiliation: Research Centre for Reproductive Health, The Robinson Institute, School of Paediatrics and Reproductive Health, The University of Adelaide, 2nd Floor, Medical School South, Adelaide, SA 5005, Australia.
authors.forename: Jeremy G
authors.initials: JG
authors.lastname: Thompson
authors.affiliation: Area of Biochemistry, Institute of Research and Technology on Animal Reproduction, School of Veterinary Sciences, University of Buenos Aires, Chorroarín 280, Buenos Aires C1427CWO, Argentina.
authors.forename: Pablo D
authors.initials: PD
authors.lastname: Cetica
mesh_terms: D015120:6-Aminonicotinamide / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D054885:Cumulus Cells / Q000502:physiology; D005260:Female; D005947:Glucose / Q000378:metabolism; D059471:In Vitro Oocyte Maturation Techniques; D019344:Lactic Acid / Q000096:biosynthesis; D008540:Meiosis / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology / Q000648:ultrastructure; D009249:NADP / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology* / Q000648:ultrastructure; D010084:Oxidation-Reduction; D010427:Pentose Phosphate Pathway / Q000187:drug effects / Q000502:physiology*
chemical_list: D002843:Chromatin; D015120:6-Aminonicotinamide; D019344:Lactic Acid; D009249:NADP; D005947:Glucose