Dataset Information

The Effect of Melatonin on Maturation, Glutathione Level and Expression of H MGB1 Gene in Brilliant Cresyl Blue (BCB) Stained Immature Oocyte.

ABSTRACT:

Objective

Nutrients and antioxidants in the medium of immature oocyte have a profound effect on maturation, fertilization and development of resulting embryos. In this study the effects of melatonin as an antioxidant agent on maturation, glutathione level and expression of High mobility group box-1 (HMGB1) gene were evaluated in immature oocytes of mice stained with brilliant cresyl blue (BCB).

Materials and methods

In this experimental study, immature oocytes were harvested from ovaries of Naval Medical Research Institute (NMRI) mice. Oocytes were stained with 26 μM BCB for 90 minutes and transferred to in vitro maturation medium containing varying doses of melatonin (10-12, 10-9, 10-6, 10-3 M) and without melatonin, for 22-24 hours. Maturation was monitored using an inverted microscope. Glutathione was assessed by monochlorobimane (MCB) staining and HMGB1 expression in mature oocyte was analyzed using real-time polymerase chain reaction (PCR).

Results

Melatonin in the concentration of 10-6 M had the most effect on maturation and HMGB1 expression of BCB+ oocytes (p<0.05). Meanwhile melatonin had no effects on glutathione levels. Additionally in immature BCB- oocytes, compared to the control group, melatonin did not affect cytoplasm maturation (p>0.05).

Conclusion

In vitro treatment with melatonin increases the maturation and HMGB1 expression in BCB+ immature oocytes and has no significant effect on glutathione levels.

SUBMITTER: Salimi M

PROVIDER: S-EPMC3866532 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Publications

The Effect of Melatonin on Maturation, Glutathione Level and Expression of H MGB1 Gene in Brilliant Cresyl Blue (BCB) Stained Immature Oocyte.

Salimi Maryam M Salehi Mohammad M Masteri Farahani Reza R Dehghani Maryam M Abadi Mohammad M Novin Marefat Ghaffari MG Nourozian Mohsen M Hosseini Ahmad A

Cell journal 20131120 4

<h4>Objective</h4>Nutrients and antioxidants in the medium of immature oocyte have a profound effect on maturation, fertilization and development of resulting embryos. In this study the effects of melatonin as an antioxidant agent on maturation, glutathione level and expression of High mobility group box-1 (HMGB1) gene were evaluated in immature oocytes of mice stained with brilliant cresyl blue (BCB).<h4>Materials and methods</h4>In this experimental study, immature oocytes were harvested from ...[more]

PMID: 24381853

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Project description:In briefOxidative stress due to high-fat content damages canine oocytes during long period of in vitro maturation and compromises their developmental competence. This paper shows how melatonin protects the oocytes and increases development to metaphase II stage. Results from this study would be applicable to conservation of endangered and other animal species.AbstractUnlike other domesticated animals, in vitro maturation of canine oocytes results in poor nuclear maturation to the metaphase II stage and high oocyte degeneration. The high-fat content of canine oocytes is the likely cause; it predisposes them to oxidative stress and deleterious reactive oxygen species (ROS). Melatonin (MTN) (potentially acting as a powerful antioxidant) was reported to support in vitro cultured oocytes in other species. In this work, canine cumulus oocyte complexes (COCs) were collected after routine ovariohysterectomy. Immunocytochemistry for expression of melatonin receptors (MTNR-1A and 1B) revealed that both receptors were highly expressed in canine oocytes and there was lower expression in the cumulus cells. Canine COCs matured in vitro in melatonin-supplemented culture at 100 nM concentration in low (5%) O2 incubator had a lower percentage at GV stage (6.7% ± 4.2 vs 19.8% ± 3), a higher MII stage (32.3% ± 6.4 vs 15.81% ± 8.1), lower degeneration (20.5% ± 3.2 vs 45.2% ± 5.15) and higher meiotic resumption (GVBD-MII; 56.2% ± 8.6 vs 19% ± 3), and produced lower ROS (determined after DCHFDA staining) (all P < 0.005) than oocytes cultured in high O2 (20%) incubator. The expression of ROS-regulating genes GPX-1 and catalase was significantly reduced in oocytes cultured with melatonin in low O2 compared with high O2 (P < 0.05). Importantly, the oocyte maturation rate increased significantly when G-IVF PLUS, a commercial culture medium used in human oocyte culture, was supplemented with MTN (P < 0.05). These results support the main objective of this study to analyze the beneficial effects of melatonin and low oxygen tension on both health and the developmental competence of in vitro matured canine oocytes.

Dataset Information

The Effect of Melatonin on Maturation, Glutathione Level and Expression of H MGB1 Gene in Brilliant Cresyl Blue (BCB) Stained Immature Oocyte.

Objective

Materials and methods

Results

Conclusion

Publications

The Effect of Melatonin on Maturation, Glutathione Level and Expression of H MGB1 Gene in Brilliant Cresyl Blue (BCB) Stained Immature Oocyte.

Similar Datasets

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