The Effect of Melatonin on Mitochondrial Function and Autophagy in In Vitro Matured Oocytes of Aged Mice


Zahraa Nasheed Hamad Almohammed, Ph.D, 1,2Fatemeh Moghani-Ghoroghi, Ph.D, 3Iraj Ragerdi-Kashani, Ph.D., 3Rouhollah Fathi, Ph.D., 4Leila Sadat Tahaei, M.Sc., 4Mohamad Naji, Ph.D., 5Parichehr Pasbakhsh, Ph.D., 3,*
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, International Campus, Tehran, Iran
Department of Gynecology, Alshatra Hospital, Thiqar Health Office, Health Ministry of Iraq
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Urology and Nephrology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, International Campus, Tehran, Iran
Department of Gynecology, Alshatra Hospital, Thiqar Health Office, Health Ministry of Iraq
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
Urology and Nephrology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
*Corresponding Address: P.O.Box: 14155-6447 Department of Anatomy School of Medicine Tehran University of Medical Sciences Tehran Iran Email:pasbakhsh@hotmail.com
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Abstract

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.