A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes

CEVIK, MESUT; AKKOÇ, Tolga; Bagis, haydar; TAŞ, Arzu; ARAT, Sezen

A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes

Mesut ÇEVİK, (Ondokuz Mayıs Üniversitesi, Veteriner Fakültesi, Samsun, Türkiye)

Arzu TAŞ, (TÜBİTAK MAM Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü, Kocaeli, Türkiye)

Tolga AKKOÇ, (TÜBİTAK MAM Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü, Kocaeli, Türkiye)

Haydar BAĞIŞ, (TÜBİTAK MAM Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü, Kocaeli, Türkiye)

Sezen ARAT (TÜBİTAK MAM Gen Mühendisliği ve Biyoteknoloji Araştırma Enstitüsü, Kocaeli, Türkiye)

Turkish Journal of Veterinary and Animal Sciences

1 0

Yıl: 2009 Cilt: 33 Sayı: 5 Sayfa Aralığı: 393 - 399 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes

Öz:

Bu çalışmanın amacı, in vitro mature edilmiş sığır oositlerinin elektriksel uyarım ve kimyasal ajanlar ile aktive edilmiş partenogenetik aktivasyonu ile standart IVF protokolünün, embriyo gelişim oranları yönünden karşılaştırılmasıdır. Bu amaçla, oositler 39 °C ve % 5 $CO_2$’li ortamda Tissue Culture Medium–199 (TCM–199)’da mature edildi. IVF işlemi, mature oositlerin modifiye edilmiş Tyrode-Lactate Medium’unda fertilize edilmesi ile yapıldı. Partenogenetik aktivasyonda, oositler 25 mikrosaniye süreyle elektriksel uyarıma (133 V/500 μm) tabi tutuldu ve daha sonra sırayla 10 dakika calcium ionophore (CaI), 1 saat cycloheximide (CHX) + cytochalasin D karışımı ve son olarak da 5 saat süreyle CHX ajanlarında tutulmak suretiyle aktivasyonları tamamlandı. Tüm embriyolar (IVF ve partenogenetik) G1.3/G2.3 mediumunda ve 39 °C’de % 6 $CO_2$, % 5 $O_2$ ve % 89 $N_2$ gaz karışımı ortamında kültüre edilmişlerdir. Partenogenetik aktivasyon ve IVF sonrası bölünme oranı yönünden yapılan karşılaştırmada önemli bir farklılık saptanmamıştır (P > 0,05). Morula, blastosist oranları ve blastosist hücre sayıları, IVF ve partenogenetik aktivasyon uygulamaları için karşılaştırıldığında aynı şekilde fark önemsiz bulundu (P > 0,05). Bu sonuçlar göstermiştir ki, sığır oositlerinin aktivasyonunda kimyasal ajanlar ile kombine edilmiş elektrik stimülasyonu uygulaması arzu edilen sonuçlar vermektedir ve kullanılan kültür mediumu (G1.3/G2.3) hem partenogenetik ve hem de IVF embriyolarının gelişimini desteklemiştir.

Anahtar Kelime: embriyo sığır oosit olgunlaşması oosit inek partenogenez embriyonik gelişim sikloheksimit in vitro döllenme blastosist

Konular: Ziraat Mühendisliği

İn vitro mature edilmiş sığır oositlerinin partenogenetik aktivasyonu ve in vitro fertilizasyonunun karşılaştırmalı çalışması

Öz:

The objective of this study was to determine the effects of 3 chemical agents used sequentially and electrical stimuli on parthenogenetic activation of in vitro matured bovine oocytes and comparison with a standard IVF protocol for embryo developmental rates. For this purpose, oocytes were matured in tissue culture medium-199 (TCM-199) at 39 °C and 5% of $CO_2$ in humidified air. For IVF, matured oocytes were fertilized in Modified Tyrode-Lactate Medium. In the parthenogenetic activation process, direct current (DC) was pulsed (133 V/500 μm) for 25 μs and oocytes were sequentially activated with calcium ionophore (CaI) for 10 min, cycloheximide (CHX) + cytochalasin D for 1 h, and CHX for 5 h. After that, all embryos (both IVF and parthenogenetic) were cultured in G1.3/G2.3 media containing 6% $CO_2$, 5% $O_2$, and 89% $N_2$ in humidified air at 39 °C. Cleavage rate was not significantly different following parthenogenetic activation compared to IVF (P > 0.05). Morula, blastocyst development, and blastocyst cell numbers were not also significantly different between parthenogenetic activation and IVF (P > 0.05). These results showed that bovine oocyte activation with electrical stimulation and chemical agents gave the desirable results, and the culture medium (G1.3/G2.3) supported both parthenogenetic and IVF embryo development.

Anahtar Kelime: cycloheximide in vitro fertilization blastocyst embryos cattle oocyte maturation oocytes cows parthenogenesis embryonic development

Konular: Ziraat Mühendisliği

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

1. Yi, Y.J., Park, C.S.: Parthenogenetic development of porcine oocytes treated by ethanol, cycloheximide, cytochalasin B and 6-dimethylaminopurine. Anim. Reprod. Sci., 2005; 86: 297-304.
2. Bhak, J-S., Lee, S-L., Ock, S-A., Mohana Kumar, B., Choe, S-Y., Rho, G-J.: Developmental rate and ploidy of embryos produced by nuclear transfer with different activation treatments in cattle. Anim. Reprod. Sci., 2006; 92: 37-49.
3. Grabiec, A., Max, A., Tischner, M.: Parthenogenetic activation of domestic cat oocytes using ethanol, calcium ionophore, cycloheximide and a magnetic field. Theriogenology, 2007; 67: 795-800.
4. Alberio, R., Zakhartchenko, V., Motlik, J., Wolf, E.: Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. Int. J. Dev. Biol., 2001; 45: 797-809.
5. Booth, P.J., Holm, P., Vajta, G., Greve, T., Callesen, H.: Effect of two activation treatments and age of blastomere karyoplasts on in vitro development of bovine nuclear transfer embryos. Mol. Reprod. Dev., 2001; 60: 377-383.
6. Tosti, E., Boni, R., Cuomo, A.: Fertilization and activation currents in bovine oocytes. Reproduction, 2002; 124: 835-846.
7. Méo, S.C., Leal, C.L.V., Garcia, J.M.: Activation and early parthenogenesis of bovine oocytes treated with ethanol and strontium. Anim. Reprod. Sci., 2004; 81: 35-46.
8. Shen, P.C., Lee, S.N., Liu, B.T., Chu, F.H., Wang, C.H., Wu, J.S., Lin, H.H., Cheng, W.T.K.: The effect of activation treatments on the development of reconstructed bovine oocytes. Anim. Reprod. Sci., 2008; 106: 1-12.
9. Arat, S., Bağış, H., Taş, A., Akkoç, T.: Effect of several parameters on parthenogenetic bovine embryo development in vitro. Reprod. Fert. Develop., 2006; 18: 119.
10. Nánássy, L., Lee, K., Jávor, A., Macháty, Z.: Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos. Anim. Reprod. Sci., 2008; 104: 264-274.
11. Wang, Z-G., Wang, W., Yu, S-D., Xu, Z-R.: Effects of different activation protocols on preimplantation development, apoptosis and ploidy of bovine parthenogenetic embryos. Anim. Reprod. Sci., 2008; 105: 292-301.
12. Gasparrini, B., Boccia, L., De Rosa, A., Di Palo, R., Campanile, G., Zicarelli, L.: Chemical activation of buffalo (Bubalus bubalis) oocytes by different methods: effects of aging on postparthenogenetic development. Theriogenology, 2004; 62: 1627- 1637.
13. Hwang, S., Lee, E., Yoon, J., Yoon, B-K., Lee, J-H., Choi, D.: Effects of electric stimulation on bovine oocyte activation and embryo development in intracytoplasmic sperm injection procedure. J. Assist. Reprod. Genet., 2000; 17: 310-314.
14. Arat, S., Bağış, H., Mercan Odaman, H., Dinnyés, A.: Cloned embryos can be produced using donor cells obtained from a 72- hour cooled carcass. Reprod. Fert. Develop., 2005; 17: 164.
15. Parrish, J.J., Susko-Parrish, J.L., Leibfried-Rutledge, M.L., Critser E.S., Eyestone, W.H., First, N.L.: Bovine in vitro fertilization with frozen-thawed semen. Theriogenology, 1986; 25: 591-600.
16. Keskintepe, L., Brackett, B.G.: In vitro developmental competence of in vitro matured bovine oocytes fertilized and cultured in completely defined media. Biol. Reprod., 1996; 55: 333-339.
17. Parrish, J.J., Krogenaes, A., Susko-Parrish, J.L.: Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development. Theriogenology, 1995; 44: 859-869.
18. Sağırkaya, H., Yağmur, M., Nur, Z., Soylu, M.K.: Replacement of fetal calf serum with synthetic serum substitute in the in vitro maturation medium: effects on maturation, fertilization and subsequent development of cattle oocytes in vitro. Turk. J. Vet. Anim. Sci., 2004; 28: 779-784.
19. Arat, S., Gibbons, J., Rzucidlo, S.J., Respess, D.S., Tumlin, M., Stice, S.L.: In vitro development of bovine nuclear transfer embryos from transgenic clonal lines of adult and fetal fibroblast cells of the same genotype. Biol. Reprod., 2002; 66: 1768-1774.
20. SAS: SAS Statistical Software Program, 2007; SAS User Guide Ver: 9.01. SAS Campus Drive, Cary NC 27513 USA.
21. Saikhun, J., Kitiyanant, N., Songtaveesin, C., Pavasuthipaisit, K., Kitiyanant, Y.: Development of swamp buffalo (Bubalus bubalis) embryos after parthenogenetic activation and nuclear transfer using serum fed or starved fetal fibroblasts. Reprod. Nutr. Dev., 2004; 44: 65-78.
22. Liu, L., Ju, J-C., Yang, X.: Parthenogenetic development and protein patterns of newly matured bovine oocytes after chemical activation. Mol. Reprod. Dev., 1998; 49: 298-307.
23. Dinnyés, A., Dai, Y., Jiang, S., Yang, X.: High developmental rates of vitrified bovine oocytes following parthenogenetic activation, in vitro fertilization, and somatic cell nuclear transfer. Biol. Reprod., 2000; 63: 513-518.

APA	CEVIK M, TAŞ A, AKKOÇ T, Bagis h, ARAT S (2009). A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. , 393 - 399.
Chicago	CEVIK MESUT,TAŞ Arzu,AKKOÇ Tolga,Bagis haydar,ARAT Sezen A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. (2009): 393 - 399.
MLA	CEVIK MESUT,TAŞ Arzu,AKKOÇ Tolga,Bagis haydar,ARAT Sezen A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. , 2009, ss.393 - 399.
AMA	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. . 2009; 393 - 399.
Vancouver	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. . 2009; 393 - 399.
IEEE	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S "A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes." , ss.393 - 399, 2009.
ISNAD	CEVIK, MESUT vd. "A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes". (2009), 393-399.

APA	CEVIK M, TAŞ A, AKKOÇ T, Bagis h, ARAT S (2009). A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. Turkish Journal of Veterinary and Animal Sciences, 33(5), 393 - 399.
Chicago	CEVIK MESUT,TAŞ Arzu,AKKOÇ Tolga,Bagis haydar,ARAT Sezen A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. Turkish Journal of Veterinary and Animal Sciences 33, no.5 (2009): 393 - 399.
MLA	CEVIK MESUT,TAŞ Arzu,AKKOÇ Tolga,Bagis haydar,ARAT Sezen A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. Turkish Journal of Veterinary and Animal Sciences, vol.33, no.5, 2009, ss.393 - 399.
AMA	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. Turkish Journal of Veterinary and Animal Sciences. 2009; 33(5): 393 - 399.
Vancouver	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes. Turkish Journal of Veterinary and Animal Sciences. 2009; 33(5): 393 - 399.
IEEE	CEVIK M,TAŞ A,AKKOÇ T,Bagis h,ARAT S "A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes." Turkish Journal of Veterinary and Animal Sciences, 33, ss.393 - 399, 2009.
ISNAD	CEVIK, MESUT vd. "A comparative study of parthenogenetic activation and in vitro fertilization of in vitro matured bovine oocytes". Turkish Journal of Veterinary and Animal Sciences 33/5 (2009), 393-399.