PARTHENOGENETIC ACTIVATION OF SHEEP OOCYTES

Year 2008, Volume: 34 Issue: 2, 1 - 5, 21.11.2012

Ümüt Cirit Kamber Demir Süleyman Bacınoğlu Elif Karaman Serhat Pabuccuoğlu , Sema Birler

Abstract

ABSTRACT

In order to establish somatic- and embryonic-cell nuclear transfer procedures in sheep in our laboratory, two activation protocols for parthenogenetic activation of sheep oocytes were tested. Oocytes obtained from ovaries of slaughtered ewes were matured for 24 h in medium 199 supplemented with pyruvate, hormones (FSH and LH) and 10% FCS. After maturation all oocytes were exposed to single electrical stimulation of 1.25 kV/cm for 80 µsec, then placed into ionomycine and cytochalasin B for 5 min and 30 min respectively, and then divided into 2 groups randomly. While oocytes in Group I were incubated in 6-Dimethylaminopurine (6-DMAP) for 2 h, oocytes in Group II were kept in cycloheximide (CHX) for 3 h before transferring into culture medium. Cleavage and blastocyst rates in groups I and II were 82.7% and 81.3%; and 2.3% and 0% respectively (P>0.05). These are our preliminary results for the parthenogenetic development of sheep oocytes in Turkey and represent promising results in the direction to achieve sheep cloning.

Key words: Parthenogenetic Activation, Sheep, Oocyte

 

 

KOYUN OOSİTLERİNİN PARTENOGENETİK AKTİVASYONU

ÖZET

Koyunlarda somatik ve embriyonik hücre nüklear transfer yöntemlerini laboratuvarımızda başlatabilmek için, iki farklı partenogenetik aktivasyon protokolü denendi. Oositler mezbahada kesilen koyunların ovaryumlarından elde edildi ve piruvat, hormonlar (FSH ve LH) ve %10 FCS katkılı medyum 199 içerisinde 24 saat olgunlaştırıldı. Olgunlaştırılan oositlere 80 µsec süreyle 1.25 kV/cm elektrik akımı verildi ve oositlerin tamamı önce 5 dk ionomisin içerisinde, daha sonra ise 30 dk sitokalasin B içerisinde bırakıldı. İki gruba rastgele ayrılan oositler, 1. grupta 6-DMAP'lı medyumda 2 saat, 2. grupta ise sikloheksimid'li medyumda 3 saat inkübe edildikten sonra kültür medyumuna aktarıldı. Grup I ve II'de elde edilen embryolarda yarıklanma oranları sırasıyla %82.7 ve %81.3 olurken blastosist oranları %2.3 ve %0 olarak bulundu (P>0.05). Sunulan çalışma koyunların klonlaması konusunda önemli aşamalardan birini oluşturan partenogenetik aktivasyon konusunda ülkemizde gerçekleştirilmiş ilk çalışmadır.

Anahtar kelimeler: Koyun, Oosit, Partenogenetik aktivasyon

Keywords

Koyun, Oosit, Partenogenetik aktivasyon

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