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Effect of culture media on porcine embryos produced by in vitro fertilization or parthenogenetic activation after oocyte maturation with cycloheximide

Published online by Cambridge University Press:  14 January 2011

Mariana Groke Marques ,
Anibal Ballarotti Nascimento ,
Renato Pereira da Costa Gerger ,
José Sergio de Arruda Gonçalves ,
Ana Rita de Sousa Coutinho ,
Renata Simões ,
Mayra Elena Ortiz D'Avila Assumpção  and
José Antônio Visintin
Mariana Groke Marques*
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva no. 87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
Anibal Ballarotti Nascimento
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
Renato Pereira da Costa Gerger
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
José Sergio de Arruda Gonçalves
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
Ana Rita de Sousa Coutinho
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
Renata Simões
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
Mayra Elena Ortiz D'Avila Assumpção
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
José Antônio Visintin
Affiliation:
Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva n°87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil.
*
All correspondence to: Mariana Groke Marques. Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences of the University of São Paulo. São Paulo–SP, Brazil, Rua Orlando Marques de Paiva no. 87, CEP 05508270 – Cidade Universitaria, São Paulo – SP, Brazil. Tel: +55 11 30917916. Fax: +55 11 30917412. e-mail: marigroke@gmail.com
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Summary

This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 μg/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX– blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.

Keywords

cycloheximidein vitro fertilizationoocyte maturationparthenogenetic activationswine
Type
Research Article
Information
Zygote , Volume 19 , Issue 4 , November 2011 , pp. 331 - 337
Copyright
Copyright © Cambridge University Press 2011

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References

Abeydeera, L.R. (2002). In vitro production of embryos in swine. Theriogenology 57, 257–73.Google Scholar
Beux, G.L., Richard, J. & Sirard, MA. (2003). Effect of cycloheximide, 6-DMAP, roscovitine and butyrolactone I on resumption of meiosis in porcine oocytes. Theriogenology 60, 1049–58.Google Scholar
Bing, Y., Che, L., Hirao, Y., Takenouchi, N., Rodrigues-Martinez, H. & Nagai, T. (2003). Parthenogenetic activation and subsequent development of porcine oocytes activated by a combined electric pulse and butyrolactone I treatment. J. Reprod. Dev. 49, 159–66.CrossRefGoogle ScholarPubMed
Booth, P.J., Holm, P. & Callesen, H. (2005). The effect of oxygen tension on porcine embryonic development is dependent on embryo type. Theriogenology 63, 2040–52.Google Scholar
Coy, P., Romar, R. (2002). In vitro production of pig embryos: a point of view. Reprod. Fertil. Dev. 14, 275–86.CrossRefGoogle ScholarPubMed
Goissis, M.D., Caetano, H.V.A., Marques, M.G., Barros, F.R.O., Feitosa, W.B., Milazzotto, M.P., Binelli, M., Assumpção, M.E.O.A. & Visintin, J.A. (2007). Effects of serum deprivation and cycloheximide on cell cycle of low and high passage porcine fetal fibroblasts. Reprod. Dom. Anim. 42, 660–3.CrossRefGoogle ScholarPubMed
Grupen, C.G., Nagashima, H. & Nottle, M.B. (1997). Asynchronous meiotic progression in porcine oocytes matured in vitro: a cause of polyspermic fertilization? Reprod. Fertil. Dev. 9, 187–91.CrossRefGoogle ScholarPubMed
Hashimoto, S., Kimura, K., Iwata, H. & Takakura, R. (2003). Oocyte transport: developmental competence of bovine oocytes arrest at germinal vesicle stage by cycloheximide under air. J. Reprod. Dev. 49, 61–6.CrossRefGoogle ScholarPubMed
Im, G.S., Liangxue, L., Liu, Z., Hao, Y., Wax, D., Bonk, A. & Prather, R.S. (2004). In vitro development of preimplantation porcine nuclear transfer embryos cultured in different media and gas atmospheres. Theriogenology 61, 1125–35.CrossRefGoogle ScholarPubMed
Kubelka, M., Motlík, J., Schultz, R.M. & Pavlok, A. (2000). Butyrolactone I reversibly inhibits meiotic maturation of bovine oocytes, without influencing chromosome condensation activity. Biol. Reprod. 62, 292–302.Google Scholar
Leal, C.L.V. & Liu, L. (1998). Differential effects of kinase inhibitor and electrical stimulus on activation and histone H1 kinase activity in pig oocytes. Anim. Rep. Sci. 52, 51–61.CrossRefGoogle ScholarPubMed
Lonergan, P., Fair, T., Khatir, H., Cesaroni, G. & Mermillod, P. (1998). Effect of protein synthesis inhibition before or during in vitro maturation on subsequent development of bovine oocytes. Theriogenology 50, 417–31.CrossRefGoogle ScholarPubMed
Marques, M.G., Nicacio, A.C., Oliveira, V.P., Nascimento, A.B., Caetano, H.V.A., Mendes, C.M., Mello, M.R.B., Milazzotto, M.P., Assumpção, M.E.O.A. & Visintin, J.A. (2007). In vitro maturation of pig oocytes with different media, hormone and meiosis inhibitors. Anim. Rep. Sci. 97, 375–81.CrossRefGoogle ScholarPubMed
Nagai, T. (1994). Current status and perspectives in IVM-IVF of porcine oocytes. Theriogenology 41, 73–8.CrossRefGoogle Scholar
Pavlok, A., Kanka, J., Motlik, J. & Vodicka, P. (2000). Culture of bovine oocytes from small antral follicles in meiosis-inhibiting medium with butyrolactone I: RNA synthesis, nucleolar morphology and meiotic competence. Anim. Rep. Sci. 64, 1–11.CrossRefGoogle ScholarPubMed
Saeki, K., Nagao, Y., Kishi, M. & Nagai, M. (1997). Developmental capacity of bovine oocytes following inhibition of meiotic resumption by cycloheximide or 6 dimethylaminopurine. Theriogenology 48, 1161–72.CrossRefGoogle ScholarPubMed
Saeki, K., Nagao, Y., Kishi, M. & Iritani, A. (1998). Timing of completion of the first meiotic division in bovine oocytes after maintenance of meiotic arrest with cyclohexamide and their subsequent development. J. Vet. Med. Sci. 60, 523–6.Google Scholar
Ye, J., Flint, P.F., Campbell, H.S. & Luck, M.R. (2002). Synchronization of porcine oocyte meiosis using cyclohexamide and its application to the study of regulation by cumulus cells. J. Reprod. Dev. 14, 433–42.Google Scholar
Ye, J., Campbell, J. & Luck, M.R., (2005). Dynamic chances in meiotic progression and improvement of developmental competence of pig oocytes in vitro by follicle stimulating hormone and cycloheximide. Biol. Reprod. 72, 399–406.CrossRefGoogle Scholar
Yi, Y.J. & Park, C.S. (2005). Parthenogenetic development of porcine oocytes treated by ethanol, cycloheximide, cytochalasin B and 6-dimethylaminopurine. Anim. Rep. Sci. 86, 297–304.CrossRefGoogle ScholarPubMed
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M.K. & Iwamura, S. (2002). Birth of piglets derived from porcine zygotes culture in chemically defined medium. Biol. Reprod. 66, 112–9.CrossRefGoogle ScholarPubMed
Zhu, J., Telfer, E.E., Fletcher, J., Springbett, A., Dobrinsky, J.R., De Souza, P.A. & Wilmut, I. (2002). Improvement of electrical activation protocol for porcine oocytes. Biol. Reprod. 66, 635–41.Google Scholar