Although there are many reports on the effect of glucose metabolism on oocyte nuclear maturation, there are few studies on its effect on ooplasmic maturation. By manipulating glucose metabolism pathways using a maturation medium that could support oocyte nuclear maturation but only a limited blastocyst formation without glucose, this study determined effects of glucose metabolism pathways on ooplasmic maturation. During maturation of cumulus-oocyte-complexes (COCs) with glucose, the presence of PPP inhibitor, DHEA or glycolysis inhibitor, iodoacetate significantly decreased blastocyst rates, intraoocyte glutathione and ATP. While blastocyst rates, GSH/GSSG ratio and NADPH were higher, ROS was lower significantly in COCs matured with iodoacetate than with DHEA. Fructose-6-phosphate overcame the inhibitory effect of DHEA on PPP. During maturation of COCs with pyruvate, electron transport inhibitor, rotenone or monocarboxylate transfer inhibitor, 4-CIN significantly decreased blastocyst rates. Cumulus-denuded oocytes had a limited capacity to use glucose or lactate, but they could use pyruvate to support maturation. In conclusion, whereas glycolysis promoted ooplasmic maturation mainly by supplying energy, PPP facilitated ooplasmic maturation to a greater extent by both reducing oxidative stress and supplying energy through providing fructose-6-phosphate for glycolysis. Pyruvate was transferred by monocarboxylate transporters and utilized through mitochondrial electron transport to sustain ooplasmic maturation.
Background:Gender is one of the risk factors accounting for the high prevalence of adolescent myopia. Considerable research results have shown that myopia incidence of female is higher than that of male. This study aimed to analyze the correlation between ocular parameters and serum estrogen level and to investigate the vision changes along with estrogen change in menstrual cycle of adolescent females.Methods:A total of 120 young females aged between 15 and 16 years, diagnosed with myopia were recruited. Spherical lens, cylindrical lens, axis, interpupillary distance (IPD), and vision in each tested eye of the same subject were measured by automatic optometry and comprehensive optometry, with repetition of all measurements in the menstrual cycle of the 2nd or 3rd days, 14th days, and 28th days, respectively. Serum estradiol (E2) levels were assayed by chemiluminescence immunoassay at the same three times points of the menstrual cycle mentioned above.Results:In young females with myopia, the spherical lens showed a statistically significant difference among all different time in menstrual cycle (all P < 0.0001). The cylindrical lens, axis, and IPD were changed significantly during the menstrual cycle (P < 0.05). The vision of the three different time points in menstrual cycle had a significant difference (χ2 = 6.35, P = 0.042). The vision during the 14th and 28th day was higher compared to that on the 2nd or 3rd days (P = 0.021). Serum E2 levels were significantly different at different time points in menstrual cycle (P < 0.05). E2 levels reached its maximum value on the 14th day and the minimum value on the 2nd or 3rd day.Conclusions:In adolescent females, the spherical lens and other related ocular parameters vary sensitively with different levels of E2 in menstrual cycle. Vision in late menstrual stage is significantly higher than that in premenstrual stage.
In previous studies on glucose metabolism during in vitro maturation, intact cumulus-oocyte complexes (COCs) were treated with enzyme inhibitors/activators. Because inhibitors/activators may have non-specificity and/or toxicity, and culture of COCs cannot differentiate whether glucose metabolism of cumulus cells (CCs) or that of the oocyte supports oocyte maturation, results from the previous studies must be verified by silencing genes in either CCs or cumulus-denuded oocytes (DOs). In this study, RNAi was adopted to specify the effects of glucose metabolism in CCs or DOs on oocyte maturation. Although silencing either glyceraldehyde 3-phosphate dehydrogenase (GAPDH) or glucose-6-phosphate dehydrogenase (G6PD) genes in CCs significantly decreased competence of the cocultured DOs, silencing G6PD impaired competence to a greater extent. While silencing G6PD or GAPDH of CCs decreased glutathione and ATP contents of cocultured DOs to similar extents, silencing G6PD increased oxidative stress as well. Analysis on metabolite contents and oxidative stress index and culture of DOs in medium conditioned with gene-silenced CCs indicated that CCs supported oocyte maturation by releasing glucose metabolites. Silencing mitochondrial pyruvate carrier 1 or NADH dehydrogenase (ubiquintone) flavoprotein 1 of DOs significantly impaired their maturation. The results have unequivocally confirmed that CCs promote oocyte maturation by releasing glucose metabolites from both pentose phosphate pathway (PPP) and glycolysis. Pyruvate is transferred into DOs by mitochondrial pyruvate carrier (MPC) and utilized through mitochondrial electron transport to support maturation.
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