Effects of Vitamin C and Melatonin on the Viability and Mitochondrial Distribution of In vitro Matured Dromedary Camel Oocytes

Document Type : Original Article

Authors

1 Dept. of animal reproduction & A.I., Veterinary Research Division, National Research Centre

2 Dept. of Theriogenology, Faculty of Veterinary Medicine, Alexandria University, Egypt.

Abstract

The purpose of this work was to look into the impact of antioxidants on the viability and mitochondrial distribution of dromedary camel oocytes, through 1) Study the effect of vitamin C (ascorbic acid) and melatonin on the maturation rate and fertilization rate of dromedary camel oocytes, 2) Study the impact of vitamin C and melatonin on the viability, mitochondrial distribution, and intensity of in-vitro matured dromedary camel oocytes. Camel oocytes were aspirated from ovaries. Oocytes of excellent and good quality were in-vitro matured using in vitro maturation media (TCM-199 + 10 ug/ml FSH + 10%fetal calf serum + 100 IU/ml Pregnant mare serum + 50 mg/ml gentamycin) without (control group) or with antioxidant groups (50 µg/ml vitamin C or 100 µM/ml melatonin) and incubated in CO2 incubator (38.5 ̊C, 5% CO2, 20% O2 and 95% humidity) for 40 hours. In vitro matured oocytes with 1st polar body were either in vitro fertilized using epididymal spermatozoa for detection the fertilization rate or stained with Hoechst 33342 (bis-Benzimide H33258) dye and MitoTracker red dye for detection the viability and mitochondrial distribution and intensity using confocal microscope. Results showed that, the nuclear maturation rate of dromedary camel oocytes in vitro matured with antioxidants were significantly (P˂0.001) higher than the control group. The fertilization rate of in vitro matured oocytes in the control group (56.26 ± 0.63) was lower than vitamin C and melatonin group (70.88 ± 0.87 and 76.63 ± 0.82, respectively). The mitochondrial intensity was significantly increased in the vitamin C (118.3 ± 1.13) and melatonin (231.1 ± 4.06) groups when compared with control group (102.5 ± 2.16). The melatonin group was significantly higher in peripheral mitochondrial distribution when compared with control and vitamin C groups, while the diffused distribution of mitochondria was markedly greater in the vitamin C and control group when compared with melatonin group. In-conclusion, Supplementation of vitamin C or melatonin on in vitro maturation media were significantly improved the maturation rate and fertilization rate through enhancing the mitochondrial intensity and distribution in Dromedary camel oocytes.

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