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Asian Journal of Animal and Veterinary Advances

Year: 2014 | Volume: 9 | Issue: 11 | Page No.: 713-720
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Authors

Sh. A. Gabr

Sh. M. Samiah

W.M. Nagy

Keywords

Research Article

Effect of Leptin Supplementation in Maturation Medium on in vitro Nuclear Maturation and Fertilization of Camel Oocytes (Camelus dromedarius)

Sh. A. Gabr, Sh. M. Samiah and W.M. Nagy
The present study aimed to evaluate the effect of leptin addition (0, 10, 20 and 30 ng mL-1) to maturation media on in vitro maturation (IVM) and fertilization (IVF) of dromedary camel oocytes. Ovaries were collected from slaughtered animals, oocytes were collected by slicing technique and matured in TCM-199 medium with different leptin levels in CO2 incubator (5% CO2) at 38.5°C and high humidity for 42 h. After maturation, oocytes were categorized as Germinal Vesicle (GV), Germinal Vesicle Breakdown (GVBD), metaphase-I (MI), metaphase-II (MII) and degenerated oocytes. Epididymal spermatozoa were recovered from camel testes immediately after slaughter. In vitro fertilization was carried out for oocytes matured in TCM-199 with 10% FDCS plus 20 μg mL-1 of leptin. Spermatozoa and oocytes were co-incubated at 38.5°C in a moisture atmosphere of 5% CO2 in air for 20-24 h. The cleaved oocytes were examined after five days of culture for cleavage stages (2, 4 and 8-16 cell, morula and blastocyst stages). Results revealed that supplementation of leptin (20 mg mL-1) showed (p<0.05) the highest percentage of oocytes at MII (58.8%) and the lowest percentages of those at GV (7.8%), GVBD (9.8%), MI (10.8%) and degenerated oocytes (12.7%) as compared to control medium (40.0, 10.9, 11.8, 14.5 and 22.7%, respectively) and other levels of leptin. Supplementation of leptin, matured with 20 ng mL-1 leptin increased fertilization rate (27 vs. 25.3%) and blastocyst rate (3.6 vs. 1.4%) as compared to the control medium. In conclusion, supplementation of leptin to maturation medium (TCM-199) at a level 20 ng mL-1 increased in vitro nuclear maturation rate of immature oocytes and their fertilization rate and blastocyst production.
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