Effect of Leptin on In Vitro Nuclear Maturation and Apoptosis of Buffalo (Bubalus bubalis) Oocyte

International Journal of Fertility and Sterility

卷:8

Gholamreza Najafi¹  Amir Khaki1²  Rouzali Batavani²  Abolfazl Belbasi²  Hamid Tahmasbian²  Aram Mokarizadeh³ 

[1] Department of Basic Sciences, Division of Embryology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran;

[2] Department of Clinical Sciences,Division of Theriogenology, Faculty of Veterinary Medicine, Urmia University,Urmia, Iran;

[3] Department of Immunology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Irann;

关键词: buffalo;    oocyte;    leptin;    maturation;    apoptosis;   

DOI  :  

来源: DOAJ

【 摘 要 】

Background: Leptin, as a 16 kDa adipokine, is a pleiotropic cytokine-like hormone thatprimarily secreted from adipose tissue. It also involves in the regulation of energy homeostasis, neuroendocrine function, immunity, lipid and glucose homeostasis, fatty acidoxidation, angiogenesis, puberty and reproduction. The aim of this study was to investigate the effects of in vitro addition of leptin to in vitro maturation (IVM) medium onbuffalo oocyte maturation and apoptosis.Materials and Methods: In this experimental study, Ovaries from apparently normalreproductive organs of slaughtered adult buffaloes (Bubalus bubalis) with unknownbreeding history were collected from Urmia Abattoir, Urmia, Iran, and were transportedimmediately to the laboratory in a thermos flask containing sterile normal saline withadded antibiotics. Oocytes were aspirated from 2-8 mm visible follicles of the ovariesusing an 18-G needle attached to a 10 ml syringe. IVM medium included tissue culturemedium-199 (TCM-199), 10% fetal bovine serum (FBS), 22 µg/ml sodium pyruvate, 0.5IU/ml ovine follicle-stimulating hormone (oFSH), 0.5 IU/ml ovine luteinizing hormone(oLH), 1 μg/ml oestradiol, 50 μg/ml gentamycin, and leptin [0 (control), 10, 50, and 100ng/ml]. The good quality buffalo oocytes (batches of 10 oocytes) were placed in a cultureplate containing six 50 μl droplets of maturation medium, covered with sterilized mineraloil, and then incubated at 38.5˚C with 5% CO2in air for 24 hours. The maturation of oocytes was evaluated under a stereomicroscope by detecting the first polar body extrusionof oocytes. FITC-Annexin V propidium iodide (PI) staining method was used to detectoocyte apoptosis. Results:From a total of 115 collected ovaries, 1100 oocytes were recovered among which283 oocyte were suitable for IVM. In the groups of leptin treated with 0 (control), 10, 50and 100 ng/ml, the percentage of oocytes maturation was 74.65, 83.81, 77.85, and 75.40%,while the percentage of oocytes apoptosis was 9.83, 9.54, 9.93, and 10.42%, respectively.Our results showed that addition of 10 ng/ml leptin to buffalo IVM medium increased oocyte maturation, significantly, as compared with that in control group. However, addition ofleptin to IVM medium had no significant influence on buffalo oocyte apoptosis.Conclusion: Our findings suggested that addition of 10 ng/ml leptin to IVM medium of buffalo oocyte can improve oocyte nuclear maturation. Furthermore, we showed that there is norelation between in vitro addition of leptin to buffalo oocyte IVM medium and oocyte apoptos Conclusion: Our findings suggested that addition of 10 ng/ml leptin to IVM medium of buffalo oocyte can improve oocyte nuclear maturation. Furthermore, we showed that there is no relation between in vitro addition of leptin to buffalo oocyte IVM medium and oocyte apoptosis.

【 授权许可】

Unknown