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Effect of Kisspeptin on In Vitro Maturation of Sheep Oocytes


Affiliations
1 Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Rajendra Nagar, Telangana, India
2 Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Korutla, Telangana, India
3 Department of Veterinary Surgery and Radiology, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Rajendra Nagar, Hyderabad, India
 

Aim: The aim of this study was to investigate the effect of kisspeptin (KP) on in vitro maturation (IVM) of sheep oocytes aspirated from the ovaries collected from slaughterhouse.
Materials and Methods: Two different experiments were conducted to investigate the effect of KP (5, 10 and 15 μg/ml) alone (experiment 1) or in combination with follicle-stimulating hormone (FSH), luteinizing hormone (LH), and Estradiol (E2) (experiment 2) on IVM of sheep oocytes. Tissue culture medium 199 supplemented with Gentamicin was used as control medium. Good quality oocytes were randomly allocated into different IVM media and cultured at 38.5°C in 5% CO2 under humidified atmosphere for 24 h. The oocytes were evaluated for their cumulus cell expansion (CCE) and extrusion of the 1st polar body (PB) at the end of maturation.
Results: The proportion of oocytes showing CCE and extrusion of PB was highest when the oocytes were matured in the medium supplemented with 10 μg/ml of KP. In experiment 2, oocytes were matured in 12 different maturation media (G1-G12: G1: Control, G2: KP alone, G3: FSH, G4: FSH+KP, G5: LH, G6: LH+KP, G7: E2, G8: E2+KP, G9: FSH+LH+E2, G10: FSH+LH+E2+KP, G11: FSH+LH+E2+fetal bovine serum (FBS), G12: FSH+LH+E2+FBS+KP). The proportion of oocytes showing cumulus expansion and PB extrusion was highest (98.33±1.05 and 89.17±2.38) when they were matured in FSH+LH+E2+FBS+KP (G12) and was significantly higher than other groups. The proportion of CCE and extrusion of PB was significantly increased when KP was supplemented to FSH and E2, but no effect was observed with LH. The maturation rates were significantly increased when FSH, LH, and E2 (G9) containing media were additionally supplemented with KP (G10).
Conclusion: This study demonstrated that the addition of KP (10 μg/ml) to the FSH, LH, and E2 supplemented media would enhance the sheep oocyte maturation in vitro.

Keywords

Kisspeptin, In Vitro Maturation, Sheep Oocytes.
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  • Effect of Kisspeptin on In Vitro Maturation of Sheep Oocytes

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Authors

Priyanka Byri
Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Rajendra Nagar, Telangana, India
Arunakumari Gangineni
Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Korutla, Telangana, India
K. Ramachandra Reddy
Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Korutla, Telangana, India
K. B. P. Raghavender
Department of Veterinary Surgery and Radiology, College of Veterinary Science, P.V. Narasimha Rao, Telangana Veterinary University, Rajendra Nagar, Hyderabad, India

Abstract


Aim: The aim of this study was to investigate the effect of kisspeptin (KP) on in vitro maturation (IVM) of sheep oocytes aspirated from the ovaries collected from slaughterhouse.
Materials and Methods: Two different experiments were conducted to investigate the effect of KP (5, 10 and 15 μg/ml) alone (experiment 1) or in combination with follicle-stimulating hormone (FSH), luteinizing hormone (LH), and Estradiol (E2) (experiment 2) on IVM of sheep oocytes. Tissue culture medium 199 supplemented with Gentamicin was used as control medium. Good quality oocytes were randomly allocated into different IVM media and cultured at 38.5°C in 5% CO2 under humidified atmosphere for 24 h. The oocytes were evaluated for their cumulus cell expansion (CCE) and extrusion of the 1st polar body (PB) at the end of maturation.
Results: The proportion of oocytes showing CCE and extrusion of PB was highest when the oocytes were matured in the medium supplemented with 10 μg/ml of KP. In experiment 2, oocytes were matured in 12 different maturation media (G1-G12: G1: Control, G2: KP alone, G3: FSH, G4: FSH+KP, G5: LH, G6: LH+KP, G7: E2, G8: E2+KP, G9: FSH+LH+E2, G10: FSH+LH+E2+KP, G11: FSH+LH+E2+fetal bovine serum (FBS), G12: FSH+LH+E2+FBS+KP). The proportion of oocytes showing cumulus expansion and PB extrusion was highest (98.33±1.05 and 89.17±2.38) when they were matured in FSH+LH+E2+FBS+KP (G12) and was significantly higher than other groups. The proportion of CCE and extrusion of PB was significantly increased when KP was supplemented to FSH and E2, but no effect was observed with LH. The maturation rates were significantly increased when FSH, LH, and E2 (G9) containing media were additionally supplemented with KP (G10).
Conclusion: This study demonstrated that the addition of KP (10 μg/ml) to the FSH, LH, and E2 supplemented media would enhance the sheep oocyte maturation in vitro.

Keywords


Kisspeptin, In Vitro Maturation, Sheep Oocytes.