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• W1963890915 abstract "Oocyte pre/postovulatory aging significantly contributes to abnormal fertilization, development and chromosome abnormalities in the embryo. The process of aging is possibly ooplasm mediated (1). However, exact mechanisms related to oocyte aging are poorly understood. Nonetheless, critical balance between the pro and antioxidant mechanisms could explain these phenomena. Hydrogen peroxide (H2O2) is an intermediate/byproduct of reactive oxygen species (ROS) metabolism, that affects oocyte Ca2+ release mechanisms at fertilization (2). On the other hand, NADPH is a cellular antioxidant. We therefore, investigate the role of H2O2 and NADPH in modulating the oxidative stress and aging related phenomena in oocytes. Mouse oocytes retrieved at 13.5 and 16.5 h after hCG were cultured for 2 h in the presence of H2O2 (group A, n=40), or microinjected with NADPH (group B, n=32), or processed without treatment (controls, group C, n=34). Aging related phenomena of zona pellucida (ZP) hardening, cortical granule (CG) loss and taxol stimulated microtubule dynamics were studied and results were compared between groups using Mann Whitney U test and Fisher's Exact test. Oocytes in group A were incubated with 20 μM H2O2 in M 16 (37°C, 5% CO2, 2 h). Oocytes from group B were microinjected with 2–4 pL of 1 μM NADPH using micromanipulation (estimated oocyte concentration 5–20 nM) and cultured (M 16, 2 h). Oocytes in group C were cultured without any treatment (2 h). All oocytes were subsequently subjected to taxol treatment (1), ZP dissolution, fluorescent α-tubulin immunocytochemistry and staining for CG prior to mounting in Vectashield with DAPI. The oocytes in individual groups were assessed for ZP dissolution time (seconds), cortical granule intactness and microtubule dynamics after taxol treatment (1) using a confocal laser scanning microscope. The ZP dissolution time (seconds) was similar in young oocytes from groups A (49.1±1.2) B (47.2±2.0) and C (46.6±1.3), but increased significantly in old oocytes from group C (66.4±1.6, P<0.001). Furthermore, H2O2 treatment resulted in significant lengthening of the ZP dissolution time in old (112.8±3.5), but not young oocytes from group A (49.1±1.2) compared to other groups (P<0.001). Increased ooplasmic microtubule dynamics were observed in old oocytes from groups A and C (80.0 and 75.0% respectively) but were remarkably diminished in all young (16/16) and most old oocytes (12/15) in group B compared to old oocytes in groups A (0/15) and C (0/15, P<0.001). Finally, the number of oocytes with intact granules was significantly higher in old oocytes from group B (80.0%) compared to old oocytes in groups A (20.0%) and C (25.0%, P<0.001). Overall, H2O2 treatment enhanced ZP hardening, ooplasmic microtubule dynamics and CG loss in old but not young oocytes, whereas NADPH microinjection seemed to protect older oocytes from these changes. Antioxidant mechanisms controlling the production/metabolism of H2O2 could be compromised in older oocytes. NADPH may play a vital role in modulating oxidative stress in oocytes. Further studies are underway to identify mechanisms involved in generation and combating oxidative stress in oocytes and their contribution to aging. References: [(1) Goud et al., Fertil Steril. 2004; 81:323–31, (2) Takahashi et al., Mol Reprod Dev. 2003; 66:143–52.]" @default.
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• W1963890915 date "2004-09-01" @default.
• W1963890915 modified "2023-10-16" @default.
• W1963890915 title "Role of NADPH and hydrogen peroxide in modulating oxidative stress and oocyte aging" @default.
• W1963890915 doi "https://doi.org/10.1016/j.fertnstert.2004.07.281" @default.
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