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Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients

Year 2010, Volume: 36 Issue: 1, 1 - 8, 13.10.2010

Abstract

ABSTRACT

The aim of this study was to obtain cloned lambs by somatic cell nuclear transfer. In vitro matured oocytes were enucleated and injected with serum-starved cumulus cells. After electrofusion and ionomycin treatments, reconstructed oocytes were activated by 10 mg/ml cycloheximide (CHX) for 5 h or 2 mM 6-dimethylaminopurine (6-DMAP) for 3 h and cultured in vitro in SOF medium at 38.5°C in a humidified atmosphere of 5% CO2, 5% O2 and 90% N2.

Cleavage rates were 18.1% (26/144) in CHX and 46.5% (140/301) in 6-DMAP groups; 11.4% of embryos developed to the morula stage in the 6-DMAP group. To investigate the developmental capacity of the embryos to become live young, early cleavage stage embryos (n= 69) were transferred to the oviducts of synchronized recipients. Five out of eight ewes (62.5%) were diagnosed as pregnant at Day 18 according to progesterone assays, and two ewes (25.0%) were diagnosed as pregnant after 45 days according to ultrasound examinations. Two healthy lambs were born by Cesarean section.

In conclusion, although by using different activation protocols blastocyst stage embryos were not obtained after in vitro culture of the somatic cell nuclear transfer embryos, live young that were born after transfer of early stage embryos showed that these embryos possess developmental capacity.

Key Words: sheep, somatic cell nuclear transfer, early transfer

References

  • Alberio, R., Zakhartchenko, V., Motlik, J . , Wolf, E . , 2001. Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. Int. J. Dev. Biol. 45, 797-809.
  • Alexander, B., Coppola, G., Di Berardino, D., Rho, G.J., St John, E . , Betts, D.H., King, W.A., 2006. The effect of 6-dimethylaminopurine (6- DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and nuclear transfer embryos. Mol. Reprod. Dev. 73, 20-30.
  • Birler, S., Pabuccuoglu, S., Alkan, S., Ozdas, O.B., Atalla, H., Ileri, I.K., 2002a. Development of in vitro derived sheep embryos to the blastocyst stage. Turk J. Vet. Anim. Sci 26, 891-894.
  • Birler, S., Pabuccuoglu, S., Atalla, H., Alkan, S., Ozdas, O.B., Bacinoglu, S., Cirit, U., Zavar, I., Sonmez, M.E.C., Ileri, I . K . , 2002b. Transfer of in vitro produced sheep embryos. Turk J. Vet. Anim. Sci. 26, 1421-1426.
  • Campbell, K.H.S., Fisher, P., Chen, W.C., Choi, I . , Kelly, R.D.W., Lee, J.-H., Xhu, J . , 2007. Somatic cell nuclear transfer: Past, present and future perspectives. Theriogenology 68S, S214- S231.
  • Corekci, S., Yilmaz, A., Birler, S., Pabuccuoglu, S., Atalla, H., 2004. A comparative study on the growth and survival characteristics of lambs produced by the transfer of in vitro produced (IVP) embryos. Turk J. Vet. Anim. Sci. 28, 831ı 835.
  • Georgiou, A.S., Snijders, A.P.L., Sostaric, E . , Aflatoonian, R., Vazquez, J . L . , Vazquez, J.M., Roca, J . , Martinez, E.A., Wright, P.C., Fazeli, A., 2007. Modulation of the oviductal environment by gametes. J. Proteome Res. 6, 4656-4666.
  • Huang, J.-C., Goldsby, J.S., Arbab, F., Melhem, Z., Aleksic, N., Wu, K . K . , 2004. Oviduct prostacyclin functions as a paracrine factor to augment the development of embryos. Hum. Reprod. 19 (12), 2907-2912.
  • Keefer, C.L., Baldassarre, H., Keystone, R., Wang, B., Bhatia, B., Bilodeau, A.S., Zhou, J . F . , Leduc, M., Downey, B.R., Lazaris, A., Karat/as, C.N., 2001. Generation of dwarf goat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes. Biol. Reprod. 64, 849-856.
  • Khosla, S., Dean, W., Brown, D., Reik, W., Feil, R., 2001. Culture of preimplantation mouse embryos affects fetal development and the expression of imprinted genes. Biol. Reprod. 64, 918-926.
  • King, T . J . , Dobrinsky, J.R., Zhu, J . , Finlayson, H.A., Bosma, W., Harkness, L . , Ritchie, W.A., Travers, A., McCorquodale, C., Day, B.N., Dinnyes, A., De Sousa, P.A., Wilmut, I., 2002. Embryo development and establishment of pregnancy after embryo transfer in pigs: coping with limitations in the availability of viable embryos. Reproduction 123, 507-515.
  • Loi, P., Ledda, S., Fulka, J . Jr., Cappai, P., Moor, R.M., 1998. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols. Biol. Reprod. 58, 1177-1187.
  • Loi, P., Boyazoglu, S., Fulka, J . Jr., Naitana, S., Cappai, P., 1999. Embryo cloning by nuclear transfer: Experiences in sheep. Livest. Prod. Sci. 60, 281-294.
  • Mahsoudi, B., L i , A., O'Neill, C., 2007. Assessment of the long-term and transgenerational consequences of perturbing preimplantation embryo development in mice. Biol. Reprod. 77, 889-896.
  • Peura, T.T., Vajda, G . , 2003. Comparison of established and new approaches in ovine and bovine nuclear transfer. Cloning Stem Cells 5-4, 257-277.
  • Ptak, G., Clinton, M., Tischner, M., Barboni, B., Mattioli, M., Loi, P., 2002. Improving delivery and offspring viability of in vitro-produced and cloned sheep embryos. Biol. Reprod 67, 1719ı 1725.
  • Sambrook, J . , Fritsch, E . F . , Maniatis, T., 1989. Molecular cloning: A laboratory manual (3 Volume set).
  • Sostaric, E . , Georgiou, A.S., Wong, C.H., Watson, P.F., Holt, W.V., Fazeli, A., 2006. Global profiling of surface plasma membrane proteome of oviductal epithelial cells. J. Proteome Res. 5, 3029- 3037.
  • Taggart, J.B., Hynes, R.A., Prodohol, P.A., Ferguson, A., 1992. A simplified protocol for routine total DNA isolation from salmonid fishes. J. of Fish Biol. 40, 963-965.
  • Takeda, H., 2007.Medaka Book.10.2.1.Genomic DNA extraction (embryo), February 2, http://www. shigen.nig.ac.j p/medaka/medakaboo
  • I. Hamzaoğlu, Ö.A. Öztürk, C. Özcan, Ö. Koçak, R. Kılıçarslan, G. Kaşıkçı, S. Toydemir, H. Dinç, E. Koban, I . Togan, I.K. Ileri and K. Ak
  • k/mdex.php?10.2.1%20genomic%20DNA°/o20e
  • xtraction (April 3, 2008).
  • Tveden-Nyborg, P., Peura, T.T., Hartwich, K.M., Walker, S.K., Maddox-Hyttel, P., 2005. Morphological characterization of pre- and periı implantation in vitro cultured, somatic cell nuclear transfer and in vivo derived ovine embryos. Reproduction 130, 681-694.
  • Walker, S.K., Hartwich, K.M., Seamark, R.F., 1996. The production of unusually large offspring following embryo manipulation: Concepts and challenges. Theriogenology 45 (1), 111-120.
  • Walmsley, S.E., Buckrell, B.C., Buschbeck, C., Rumph, N., Pollard, J.W., 2004. Rate of abnormalities in lambs from in vitro produced embryos transferred on Day 2 compared with Day 6 post fertilization. Theriogenology 62, 195-206.
  • Wells, D.N., Misica, P.M., Day, A.M., Tervit, H.R., 1997. Production of cloned lambs from an established embryonic cell line: A comparison between in vivo- and in vitro-matured cytoplasts. Biol. Reprod. 57, 385-393.
Year 2010, Volume: 36 Issue: 1, 1 - 8, 13.10.2010

Abstract

References

  • Alberio, R., Zakhartchenko, V., Motlik, J . , Wolf, E . , 2001. Mammalian oocyte activation: lessons from the sperm and implications for nuclear transfer. Int. J. Dev. Biol. 45, 797-809.
  • Alexander, B., Coppola, G., Di Berardino, D., Rho, G.J., St John, E . , Betts, D.H., King, W.A., 2006. The effect of 6-dimethylaminopurine (6- DMAP) and cycloheximide (CHX) on the development and chromosomal complement of sheep parthenogenetic and nuclear transfer embryos. Mol. Reprod. Dev. 73, 20-30.
  • Birler, S., Pabuccuoglu, S., Alkan, S., Ozdas, O.B., Atalla, H., Ileri, I.K., 2002a. Development of in vitro derived sheep embryos to the blastocyst stage. Turk J. Vet. Anim. Sci 26, 891-894.
  • Birler, S., Pabuccuoglu, S., Atalla, H., Alkan, S., Ozdas, O.B., Bacinoglu, S., Cirit, U., Zavar, I., Sonmez, M.E.C., Ileri, I . K . , 2002b. Transfer of in vitro produced sheep embryos. Turk J. Vet. Anim. Sci. 26, 1421-1426.
  • Campbell, K.H.S., Fisher, P., Chen, W.C., Choi, I . , Kelly, R.D.W., Lee, J.-H., Xhu, J . , 2007. Somatic cell nuclear transfer: Past, present and future perspectives. Theriogenology 68S, S214- S231.
  • Corekci, S., Yilmaz, A., Birler, S., Pabuccuoglu, S., Atalla, H., 2004. A comparative study on the growth and survival characteristics of lambs produced by the transfer of in vitro produced (IVP) embryos. Turk J. Vet. Anim. Sci. 28, 831ı 835.
  • Georgiou, A.S., Snijders, A.P.L., Sostaric, E . , Aflatoonian, R., Vazquez, J . L . , Vazquez, J.M., Roca, J . , Martinez, E.A., Wright, P.C., Fazeli, A., 2007. Modulation of the oviductal environment by gametes. J. Proteome Res. 6, 4656-4666.
  • Huang, J.-C., Goldsby, J.S., Arbab, F., Melhem, Z., Aleksic, N., Wu, K . K . , 2004. Oviduct prostacyclin functions as a paracrine factor to augment the development of embryos. Hum. Reprod. 19 (12), 2907-2912.
  • Keefer, C.L., Baldassarre, H., Keystone, R., Wang, B., Bhatia, B., Bilodeau, A.S., Zhou, J . F . , Leduc, M., Downey, B.R., Lazaris, A., Karat/as, C.N., 2001. Generation of dwarf goat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes. Biol. Reprod. 64, 849-856.
  • Khosla, S., Dean, W., Brown, D., Reik, W., Feil, R., 2001. Culture of preimplantation mouse embryos affects fetal development and the expression of imprinted genes. Biol. Reprod. 64, 918-926.
  • King, T . J . , Dobrinsky, J.R., Zhu, J . , Finlayson, H.A., Bosma, W., Harkness, L . , Ritchie, W.A., Travers, A., McCorquodale, C., Day, B.N., Dinnyes, A., De Sousa, P.A., Wilmut, I., 2002. Embryo development and establishment of pregnancy after embryo transfer in pigs: coping with limitations in the availability of viable embryos. Reproduction 123, 507-515.
  • Loi, P., Ledda, S., Fulka, J . Jr., Cappai, P., Moor, R.M., 1998. Development of parthenogenetic and cloned ovine embryos: Effect of activation protocols. Biol. Reprod. 58, 1177-1187.
  • Loi, P., Boyazoglu, S., Fulka, J . Jr., Naitana, S., Cappai, P., 1999. Embryo cloning by nuclear transfer: Experiences in sheep. Livest. Prod. Sci. 60, 281-294.
  • Mahsoudi, B., L i , A., O'Neill, C., 2007. Assessment of the long-term and transgenerational consequences of perturbing preimplantation embryo development in mice. Biol. Reprod. 77, 889-896.
  • Peura, T.T., Vajda, G . , 2003. Comparison of established and new approaches in ovine and bovine nuclear transfer. Cloning Stem Cells 5-4, 257-277.
  • Ptak, G., Clinton, M., Tischner, M., Barboni, B., Mattioli, M., Loi, P., 2002. Improving delivery and offspring viability of in vitro-produced and cloned sheep embryos. Biol. Reprod 67, 1719ı 1725.
  • Sambrook, J . , Fritsch, E . F . , Maniatis, T., 1989. Molecular cloning: A laboratory manual (3 Volume set).
  • Sostaric, E . , Georgiou, A.S., Wong, C.H., Watson, P.F., Holt, W.V., Fazeli, A., 2006. Global profiling of surface plasma membrane proteome of oviductal epithelial cells. J. Proteome Res. 5, 3029- 3037.
  • Taggart, J.B., Hynes, R.A., Prodohol, P.A., Ferguson, A., 1992. A simplified protocol for routine total DNA isolation from salmonid fishes. J. of Fish Biol. 40, 963-965.
  • Takeda, H., 2007.Medaka Book.10.2.1.Genomic DNA extraction (embryo), February 2, http://www. shigen.nig.ac.j p/medaka/medakaboo
  • I. Hamzaoğlu, Ö.A. Öztürk, C. Özcan, Ö. Koçak, R. Kılıçarslan, G. Kaşıkçı, S. Toydemir, H. Dinç, E. Koban, I . Togan, I.K. Ileri and K. Ak
  • k/mdex.php?10.2.1%20genomic%20DNA°/o20e
  • xtraction (April 3, 2008).
  • Tveden-Nyborg, P., Peura, T.T., Hartwich, K.M., Walker, S.K., Maddox-Hyttel, P., 2005. Morphological characterization of pre- and periı implantation in vitro cultured, somatic cell nuclear transfer and in vivo derived ovine embryos. Reproduction 130, 681-694.
  • Walker, S.K., Hartwich, K.M., Seamark, R.F., 1996. The production of unusually large offspring following embryo manipulation: Concepts and challenges. Theriogenology 45 (1), 111-120.
  • Walmsley, S.E., Buckrell, B.C., Buschbeck, C., Rumph, N., Pollard, J.W., 2004. Rate of abnormalities in lambs from in vitro produced embryos transferred on Day 2 compared with Day 6 post fertilization. Theriogenology 62, 195-206.
  • Wells, D.N., Misica, P.M., Day, A.M., Tervit, H.R., 1997. Production of cloned lambs from an established embryonic cell line: A comparison between in vivo- and in vitro-matured cytoplasts. Biol. Reprod. 57, 385-393.
There are 27 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

Sema Birler

Serhat Pabuccuoğlu

Kamber Demir This is me

Ümüt Cirit This is me

Elif Kahraman This is me

Süleyman Bacınoğlu This is me

Özen Özdaş This is me

Mithat Evecen

Serhat Alkan

Alper Baran

Gül Bakırer This is me

İzem Hamzaoğlu This is me

Ömer Öztürk This is me

Cenk Özcan This is me

Ömür Koçak

Ragıp Kılıçarslan

Güven Kaşıkçı

Seval Toydemir This is me

Havva Dinç This is me

Evren Koban This is me

İnci Togan This is me

İrfan İleri This is me

Kemal Ak

Publication Date October 13, 2010
Published in Issue Year 2010 Volume: 36 Issue: 1

Cite

APA Birler, S., Pabuccuoğlu, S., Demir, K., Cirit, Ü., et al. (2010). Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, 36(1), 1-8. https://doi.org/10.16988/iuvfd.12103
AMA Birler S, Pabuccuoğlu S, Demir K, Cirit Ü, Kahraman E, Bacınoğlu S, Özdaş Ö, Evecen M, Alkan S, Baran A, Bakırer G, Hamzaoğlu İ, Öztürk Ö, Özcan C, Koçak Ö, Kılıçarslan R, Kaşıkçı G, Toydemir S, Dinç H, Koban E, Togan İ, İleri İ, Ak K. Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients. iuvfd. October 2010;36(1):1-8. doi:10.16988/iuvfd.12103
Chicago Birler, Sema, Serhat Pabuccuoğlu, Kamber Demir, Ümüt Cirit, Elif Kahraman, Süleyman Bacınoğlu, Özen Özdaş, Mithat Evecen, Serhat Alkan, Alper Baran, Gül Bakırer, İzem Hamzaoğlu, Ömer Öztürk, Cenk Özcan, Ömür Koçak, Ragıp Kılıçarslan, Güven Kaşıkçı, Seval Toydemir, Havva Dinç, Evren Koban, İnci Togan, İrfan İleri, and Kemal Ak. “Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 36, no. 1 (October 2010): 1-8. https://doi.org/10.16988/iuvfd.12103.
EndNote Birler S, Pabuccuoğlu S, Demir K, Cirit Ü, Kahraman E, Bacınoğlu S, Özdaş Ö, Evecen M, Alkan S, Baran A, Bakırer G, Hamzaoğlu İ, Öztürk Ö, Özcan C, Koçak Ö, Kılıçarslan R, Kaşıkçı G, Toydemir S, Dinç H, Koban E, Togan İ, İleri İ, Ak K (October 1, 2010) Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 36 1 1–8.
IEEE S. Birler, “Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients”, iuvfd, vol. 36, no. 1, pp. 1–8, 2010, doi: 10.16988/iuvfd.12103.
ISNAD Birler, Sema et al. “Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi 36/1 (October 2010), 1-8. https://doi.org/10.16988/iuvfd.12103.
JAMA Birler S, Pabuccuoğlu S, Demir K, Cirit Ü, Kahraman E, Bacınoğlu S, Özdaş Ö, Evecen M, Alkan S, Baran A, Bakırer G, Hamzaoğlu İ, Öztürk Ö, Özcan C, Koçak Ö, Kılıçarslan R, Kaşıkçı G, Toydemir S, Dinç H, Koban E, Togan İ, İleri İ, Ak K. Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients. iuvfd. 2010;36:1–8.
MLA Birler, Sema et al. “Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients”. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, vol. 36, no. 1, 2010, pp. 1-8, doi:10.16988/iuvfd.12103.
Vancouver Birler S, Pabuccuoğlu S, Demir K, Cirit Ü, Kahraman E, Bacınoğlu S, Özdaş Ö, Evecen M, Alkan S, Baran A, Bakırer G, Hamzaoğlu İ, Öztürk Ö, Özcan C, Koçak Ö, Kılıçarslan R, Kaşıkçı G, Toydemir S, Dinç H, Koban E, Togan İ, İleri İ, Ak K. Production of Cloned Lambs: Transfer of Early Cleavage Stage Embryos to Final Recipients. iuvfd. 2010;36(1):1-8.