The Effect of Melatonin on Mitochondrial Function and Autophagy in In Vitro Matured Oocytes of Aged Mice
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Published on: April 16, 2018
Zahraa Nasheed Hamad Almohammed1,2, Fatemeh Moghani-Ghoroghi3, Iraj Ragerdi-Kashani3
1Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, International Campus, Tehran, Iran.
Objective:
This study examined the in vitro effect of melatonin on the protein synthesis of mitochondria, as well as autophagy in matured oocytes of aged mice.
Materials And Methods:
In this experimental study, germinal vesicles (GV) oocytes were collected from aged (with the age of six-months-old) and young mice (with age range of 6-8 weeks old) and then cultured in the in vitro culture medium (IVM) for 24 hours to each metaphase II (MII) oocytes and then supplemented with melatonin at a concentration of 10 μM. The culture medium of MII oocytes was devoid of melatonin. Afterward, the expression of the SIRT-1 and LC3 was assessed by immunocytochemistry. ATP-dependent luciferin-luciferase bioluminescence assay was employed for the measurement of the ATP contents. Intracellular reactive oxygen specious (ROS) was detected by DCFH-DA, and the total antioxidant capacity (TAC) level was determined by TAC assay.
Results:
The expression of SIRT-1 and LC3, as well as the measurement of the ATP content, was significantly increased in oocytes treated with melatonin compared with the oocytes receiving no treatment. Moreover, TAC was considerably higher in melatonin-treated oocytes than oocytes receiving no treatment. On the other hand, the level of ROS was significantly decreased in oocytes treated with melatonin in comparison with the untreated oocytes. The results indicated that melatonin considerably improved the development of oocytes as well.
Conclusion:
According to the data, melatonin increased mitochondrial function and autophagy via an increase in the expression of SIRT1 and LC3, as well as the ATP contents while it decreased the levels of ROS and increased TAC in oocytes derived from aged mice.
Insights
Melatonin supplementation improved aged mouse oocyte quality by enhancing mitochondrial function and autophagy. It boosted SIRT-1 and LC3 expression, increased ATP and antioxidant capacity, and reduced oxidative stress.
Area of Science:
- Reproductive Biology
- Cellular Biology
- Gerontology
Background:
- Oocyte aging is associated with impaired mitochondrial function and reduced autophagy.
- Melatonin, a potent antioxidant, may offer protective effects against age-related oocyte decline.
Purpose of the Study:
- To investigate the in vitro effects of melatonin on mitochondrial protein synthesis and autophagy in aged mouse oocytes.
- To assess melatonin's impact on oxidative stress and antioxidant capacity in these oocytes.
Main Methods:
- Oocytes from aged and young mice were cultured in vitro.
- Melatonin (10 μM) was added to the culture medium of aged oocytes.
- Expression of SIRT-1 and LC3, ATP content, reactive oxygen species (ROS), and total antioxidant capacity (TAC) were measured.
Main Results:
- Melatonin treatment significantly increased SIRT-1 and LC3 expression, and ATP content in aged oocytes.
- Melatonin significantly elevated TAC and decreased ROS levels compared to untreated aged oocytes.
- Melatonin treatment improved overall oocyte development.
Conclusions:
- Melatonin enhances mitochondrial function and autophagy in aged mouse oocytes by upregulating SIRT1 and LC3.
- Melatonin mitigates oxidative stress and boosts antioxidant capacity, improving oocyte quality in aged mice.
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