The effect of glucose on the progression of the nuclear maturation of pig oocytes.
The progression of the nuclear maturation of oocytes is a useful marker for the estimation of the subsequent developmental competence of oocytes. In this study, we examined the effect of energy substrates in an in vitro maturation medium on the progression of the nuclear maturation of oocytes. In experiment 1, the supplementation of the maturation medium with 0, 5 and 10 mM of glucose lead to increase in the total cell number of the blastocysts. In experiments 2 and 3, the maturation phase was divided into two stages (germinal vesicle (GV) stage: 0-20 h and nuclear maturation stage: 20-44 h), and the effects of glucose or pyruvate added at each stage on the kinetics of nuclear maturation were examined. The addition of glucose at the nuclear maturation stage rather than at the GV stage of maturation effected greater acceleration in the progression of nuclear maturation. However, the addition of pyruvate at both stages had the same effect on the progression of nuclear maturation was the same. In addition, when glucose was added to the medium containing pyruvate, an additive effect on the progression of nuclear maturation was observed (experiment 4). In experiment 5, the inhibitors of glucose-6-phosphate dehydrogenase (G6PD), dehydroepiandrosterone (DHEA) and 6-aminonicotinamide (6-AN) decreased the rate of the final maturation of oocytes and reduced the difference between the rates of the final maturation of oocytes cultured with glucose and those cultured with pyruvate. In the experiment 6, when the activator of G6PD, brilliant cresyle blue (BCB), was added to the maturation medium, the progression of nuclear maturation was significantly accelerated. The results of this study suggested that in addition to the role of an energy substrate, glucose or its metabolites play a role in nuclear maturation. This role was more pronounced at the second stage of maturation (transition from GV breakdown (GVBD) to M2), probably due to the metabolism of glucose via the pentose phosphate pathway (PPP) rather than the glycolysis pathway.
Sato
H
H
Tokyo University of Agriculture, Department of Animal Science, Funako 1737, Atsugi City, Kanagawa Prefecture, Japan.
Iwata
H
H
Hayashi
T
T
Kimura
K
K
Kuwayama
T
T
Monji
Y
Y
D015120:6-Aminonicotinamide / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects* / Q000502:physiology; D002478:Cells, Cultured; D003687:Dehydroepiandrosterone / Q000494:pharmacology; D005260:Female; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects*; D010078:Oxazines / Q000494:pharmacology; D010427:Pentose Phosphate Pathway / Q000502:physiology; D019289:Pyruvic Acid / Q000378:metabolism / Q000494:pharmacology; D013552:Swine / Q000378:metabolism*
D010078:Oxazines; D015120:6-Aminonicotinamide; D003687:Dehydroepiandrosterone; C006797:Brilliant Cresyl Blue; D019289:Pyruvic Acid; D005947:Glucose