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Effect of cytochalasins B and D on the developmental competence of somatic cell nuclear transfer embryos in miniature pigs

Published online by Cambridge University Press:  01 May 2008

Satoshi Sugimura*
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan. Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Manabu Kawahara
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Takuya Wakai
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Ken-ichi Yamanaka
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Hiroshi Sasada
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
Eimei Sato
Affiliation:
Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan.
*
All correspondence to Satoshi Sugimura. Laboratory of Animal Reproduction, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-amamiyamachi, Aoba-ku, Sendai 981-8555, Japan. Tel: +81 22 717 8687. Fax: +81 22 717 8687. e-mail: s-sugimura@bios.tohoku.ac.jp

Summary

In many animals, cytochalasins have generally been used as cytoskeletal inhibitors for the diploid complement retention of somatic cell nuclear transfer (SCNT) embryos. However, limited information is available on the effects of cytochalasins on the in vitro development of SCNT embryos. Hence, we compared the effects of cytochalasin B (CB) and cytochalasin D (CD) on pseudo-polar body (pPB) extrusion, cortical actin filament (F-actin) distribution in porcine parthenogenetic oocytes and in vitro development of SCNT embryos that were reconstructed using foetal fibroblasts in the G0/G1 phase derived from miniature pigs. CB (7.5 μg/ml) and CD (2.5 μg/ml) treatments effectively inhibited pPB extrusion in SCNT embryos. CB (2.5 μg/ml) treatment could not inhibit pPB extrusion and insufficiently destabilized F-actin immediately following artificial activation. In parthenogenetic oocytes treated with 2.5 μg/ml CD, normal reorganization and uniform distribution of cortical F-actin at the cytoplasmic membrane were observed at 8 h after artificial activation; this finding was similar to that of control oocytes. In contrast, parthenogenetic oocytes treated with 7.5 μg/ml CB showed non-uniform distribution of F-actin at 8 h after artificial activation. On day 5 after in vitro cultivation, the blastocyst formation rate of SCNT embryos treated with 2.5 μg/ml CD was significantly higher than that of SCNT embryos treated with 2.5 and 7.5 μg/ml CB (p < 0.05). Hence, the present findings suggest that CD is more effective than CB as the cytoskeletal inhibitor for the production of SCNT embryos in miniature pigs.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2008

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References

Beyhan, Z., Ross, P.J., Iager, A.E., Kocabas, A.M., Cunniff, K., Rosa, G.J. & Cibelli, J.B. (2007). Transcriptional reprogramming of somatic cell nuclei during preimplantation development of cloned bovine embryos. Dev. Biol. 305, 637–49.CrossRefGoogle ScholarPubMed
Brown, S.S. & Spudich, J.A. (1981). Mechanism of action of cytochalasin: evidence that it binds to actin filament ends. J. Cell Biol. 88, 487–91.CrossRefGoogle ScholarPubMed
Campbell, K.H., McWhir, J., Ritchie, W.A. & Wilmut, I. (1996). Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–6.CrossRefGoogle ScholarPubMed
Cooper, J.A. (1987). Effects of cytochalasin and phalloidin on actin. J. Cell Biol. 105, 1473–8.CrossRefGoogle ScholarPubMed
De Sousa, P.A., Dobrinsky, J.R., Zhu, J., Archibald, A.L., Ainslie, A., Bosma, W., Bowering, J., Bracken, J., Ferrier, P.M., Fletcher, J., Gasparrini, B., Harkness, L., Johnston, P., Ritchie, M., Ritchie, W.A., Travers, A., Albertini, D., Dinnyes, A., King, T.J. & Wilmut, I. (2002). Somatic cell nuclear transfer in the pig: control of pronuclear formation and integration with improved methods for activation and maintenance of pregnancy. Biol. Reprod. 66, 642–50.CrossRefGoogle ScholarPubMed
Inoue, K., Noda, S., Ogonuki, N., Miki, H., Inoue, S., Katayama, K., Mekada, K., Miyoshi, H. & Ogura, A. (2007). Differential developmental ability of embryos cloned from tissue-specific stem cells. Stem Cells 25, 1279–85.CrossRefGoogle ScholarPubMed
Kawahara, M., Mori, T., Tanaka, H. & Shimizu, H. (2002). The suppression of fragmentation by stabilization of actin filament in porcine enucleated oocytes. Theriogenology 58, 1081–95.CrossRefGoogle ScholarPubMed
Kawahara, M., Wakai, T., Yamanaka, K.Kobayashi, J., Sugimura, S., Shimizu, T., Matsumoto, H., Kim, J. H., Sasada, H. & Sato, E. (2005). Caffiene promotes premuture chromosome condensation fromation and in vitro development in porcine reconstructed embryos via a ligh level of maturation promoting factor activity during nuclear transfer. Reproduction 130, 351–7.CrossRefGoogle Scholar
Lai, L., Tao, T., Machaty, Z., Kuhholzer, B., Sun, Q.Y., Park, K.W., Day, B.N. & Prather, R.S. (2001). Feasibility of producing porcine nuclear transfer embryos by using G2/M-stage fetal fibroblasts as donors. Biol. Reprod. 65, 1558–64.CrossRefGoogle ScholarPubMed
Miyoshi, K., Saeki, K. & Sato, E. (2000). Improvement in development of porcine embryos reconstituted with cells from blastocyst-derived cell lines and enucleated oocytes by optimization of reconstruction methods. Cloning 2, 175–84.CrossRefGoogle ScholarPubMed
Onishi, A., Iwamoto, M., Akita, T., Mikawa, S., Takeda, K., Awata, T., Hanada, H. & Perry, A.C. (2000). Pig cloning by microinjection of fetal fibroblast nuclei. Science 289, 1188–90.CrossRefGoogle ScholarPubMed
Quinn, P., Barros, C. & Whittingham, D.G. (1982). Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa. J. Reprod. Fertil. 66, 161–8.CrossRefGoogle ScholarPubMed
Rampal, A.L., Pinkofsky, H.B. & Jung, C.Y. (1980). Structure of cytochalasins and cytochalasin B binding sites in human erythrocyte membranes. Biochemistry 19, 679–83.CrossRefGoogle ScholarPubMed
Sung, L.Y., Gao, S., Shen, H., Yu, H., Song, Y., Smith, S.L., Chang, C.C., Inoue, K., Kuo, L., Lian, J., Li, A., Tian, X.C., Tuck, D.P., Weissman, S.M., Yang, X. & Cheng, T. (2006). Differentiated cells are more efficient than adult stem cells for cloning by somatic cell nuclear transfer. Nat. Genet. 38, 1323–8.CrossRefGoogle ScholarPubMed
Wakayama, T. & Yanagimachi, R. (2001). Mouse cloning with nucleus donor cells of different age and type. Mol. Reprod. Dev. 58, 376–83.3.0.CO;2-L>CrossRefGoogle ScholarPubMed
Wakayama, T., Perry, A.C., Zuccotti, M., Johnson, K.R. & Yanagimachi, R. (1998). Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394, 369–74.CrossRefGoogle ScholarPubMed
Yamanaka, K.I., Sugimura, S., Wakai, T., Shoji, T., Kobayashi, J., Sasada, H. & Sato, E. (2007). Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. J. Reprod. Dev. 53, 791800.CrossRefGoogle ScholarPubMed
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M. & Iwamura, S. (2002). Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–9.CrossRefGoogle Scholar