Relationship between Nucleus Swelling and Development Competence of Bovine Cloned Embryos Reconstructed by Enucleated Oocytes with Serum-starved or Serum-fed Fetal Somatic Cells

Mokhamad Fahrudin; Ni Wayan Kurniani Karja; Tatsuyuki Suzuki

doi:10.22146/ijbiotech.7553

Relationship between Nucleus Swelling and Development Competence of Bovine Cloned Embryos Reconstructed by Enucleated Oocytes with Serum-starved or Serum-fed Fetal Somatic Cells

https://doi.org/10.22146/ijbiotech.7553

Mokhamad Fahrudin^(1*), Ni Wayan Kurniani Karja⁽²⁾, Tatsuyuki Suzuki⁽³⁾

(1) Faculty of Veterinary Medicine, Bogor Agricultural University, Bogor 16680, Indonesia
(2) Department of Reproduction and Obstetric, Faculty of Veterinary Medicine, Gadjah Mada University,Yogyakarta 55281, Indonesia
(3) Laboratory of Animal Reproduction and Biotechnology, The United Graduate School of VeterinarySciences, Yamaguchi University, Yamaguchi 753-8515, Japan
(*) Corresponding Author

Abstract

This study was conducted to examine the occurrence of nuclear remodeling (nucleus swelling) and its effects on the subsequent in vitro development of bovine embryos reconstructed by serum-starved and serum-fed somatic cells. Results from this study demonstrated that all of the reconstructed embryos that received serum-starved and serum-fed somatic cells exhibited condensed-nuclei. More than 90% of the transferred nuclei exhibited nuclear envelope breakdown and premature chromatin condensation which clearly distinct from an intact nucleus. There was no significant difference on the degree of nucleus swelling in SS-NT embryos or SF-NT embryos, indicating that either serum-starved or confluent somatic cell lines could be reprogrammed by the recipient cytoplasm environments in similar pattern. Although the fusion rate was not significantly different among the groups, the proportion of SS-NT embryos which developed to the 2- to 4-cell stage (89.7%) and to the 8- to 16-cell stage (74.7%) was significantly higher than that of SF-NT embryos. Whereas, the proportion of reconstructed embryos that developed to the morula and blastocyst stages were not significantly different among the groups. Results of these studies demonstrate that reconstructed embryos, which received either serum-starved or serum-fed confluent somatic cells, showed similar developmental competence to the blastocyst stage.

Keywords

nuclear transplantation technique-somatic cells-nucleus swelling

Full Text:

PDF

References

Campbell, K.H.S., Loi, P., Otaegui, P. J., and Wilmut, I., 1996. Cell cycle co- ordination in embryo cloning by nuclear transfer. Rev. Reprod. I, 40- 46.

Campbell, K.H.S., Ritchie, W.A., and Wilmut, I., 1993. Nuclear- cytoplasmic interactions during the first cell cycle of nuclear transfer reconstructed embryos: implications for deoxyribonucleic acid replication and development. Biol. Reprod. 49, 933-942.

Cheong, H.T., Takahashi, Y., and Kanagawa, H., 1993. Birth of mice after trans-plantation of early cell- cycle stage embryonic nuclei into enucleated oocytes. Biol. Reprod. 48, 958-963.

Cheong, H.T., Takahashi, Y., and Kanagawa, H., 1994. Relationship between nuclear remodeling and subsequent development of mouse embryonic nuclei transferred into enucleated oocytes. Mol. Reprod. Dev. 37, 138-145.

Cibelli, J.B., Stice, S.L., Golueke, P.J., Kane, J.J., Jerry, J., Blackwell, C., Ponce De Leon, A., and Robl, J.M., 1998. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science, 280, 1256-1258.

Collas, P., and Robl, J.M., 1991. Relationship between nuclear remodeling and development in nuclear transplant rabbit embryos. Biol. Reprod., 45, 455-465.

Collas, P., Balise, J.J., and Robl, J.M. 1992a. Influence of cell cycle stage of the donor nucleus on development of nuclear transplant rabbit embryos. Biol. Reprod., 46, 492-500.

Collas, P., Pinto-Correia, C., Pone De Leon, F.A., and Robl, J.M., 1992. Effect of donor cell cycle stage on chromatin and spindle morphology in nuclear transplant rabbit embryos. Biol. Reprod., 46, 501-511.

Gurdon, J.B., Partington, G.A., and De Robertis, E.M., 1976. Injected nuclei in frog oocytes: RNA synthesis and protein exchange. J. Embryol. Exp. Morph., 36, 541-553.

Fahrudin, M., Otoi, T., Suzuki, T., 2001. Developmental competence of bovine embryos reconstructed by the transfer of somatic cells derived from frozen tissues. J. Vet. Med. Sci., 63, 1151-1154.

Kubota, C., Yamakuchi, H., Todoroki, J., Mizoshita, K., Tabara, N., Barber, M., and Yang, X., 2000. Six cloned calves produced from adult fibroblast cells after long-term culture. Proc. Natl. Acad. Sci. USA., 97, 990-995.

Kues, W.A., Anger, M., Carnwath, J.W., Paul, D., Motlik, J., and Nieman, H., 2000. Cell cycle synchronization of porcine fetal fibroblasts: effects of serum deprivation and reversible cell cycle inhibitors. Biol. Reprod., 62, 412-419.

Ogura A., Inoue, K., Ogonuki, N., Noguchi, A., Takano, K., Nagano, R., Suzuki, sO., Lee, J., Ishino, F., and Matsuda, J., 2000. Production of male cloned mice from fresh, cultured, and cryopreserved immature sertoli cells. Biol. Reprod., 62, 1579-1584.

Prather, R.S., and First, N.L., 1990. Cloning embryos by nuclear transfer. J. Reprod. Fertil. (Suppl.), 41, 125-134.

Rosenkrans, Jr.C.F., Zeng, G.Q., Mcnamara, G.T., Schoff, P.K., and First, N.L., 1993. Development of bovine embryos in vitro as affected by energy substrates. Biol. Reprod. 49, 459-462.

Szollosi, D., Czolowska, R., Szollosi, M.S., and Tarkowski, A.K., 1988. Remodelling of mouse thymocyte nuclei depends on the time of their transfer into activated, homologous oocytes. J. Cell Sci., 91, 19-34.

Takano, H., Koyama, K., Kozai, C., Shimizu, S., Kato, Y., and Tsunoda, Y., 1996. Effects of cell cycle stage of donor nuclei on the development of bovine nuclear transferred embryos. J. Reprod. Dev., 42, 61-65.

Tarkowski, A.K. and Balakier, H., 1980. Nucleo-cytoplasmic interaction in cell hybrids between mouse oocytes, blastomere and somatic cells. J. Embryol. Exp. Morph., 55, 319-330.

Ushijima, H. and Eto, T., 1994. Bovine nuclear transplantation using a parthe-nogenetically activated oocytes as the recipient oocytes. J. Reprod. Dev., 40, 117-124.

Wells, D.N., Misica, P.M., Day, A.M., and Tervit, H.R., 1997. Production of cloned lambs from established embryonic cell line: a comparison between in vivo-and in vitro- matured cytoplast. Biol. Reprod., 57, 385-393.

Wells, D.N., Misica, P.M., and Tervit, H.R., 1999. Production of cloned calves following nuclear transfer with cultured adult mural granulose cells. Biol. Reprod., 60, 996-1005.

Wilmut, I., Schnieke, A.E., McWhir, J., Kind, A.J., and Campbell, K.H.S., 1997. Viable offspring derived from fetal and adult mammalian cells. Nature, 385, 810-813.

DOI: https://doi.org/10.22146/ijbiotech.7553