Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-26T15:11:18.575Z Has data issue: false hasContentIssue false
  • English
  • Français

Effect of trichostatin A on transfected donor cells and subsequent development of porcine cloned embryos

Published online by Cambridge University Press:  23 June 2010

Fu Bo ,
Liu Di ,
Fang Qing-chang ,
Ren Liang ,
Ma Hong ,
Wang Liang ,
Guo Zhen-hua  and
Li Zhong-qiu
Fu Bo
Affiliation:
College of Life Science, Northeast Agriculture University, Harbin 150030, P.R. China.
Liu Di*
Affiliation:
Heilongjiang Academy of Agricultural Science; 368 Xue Fu Road, Harbin City 150086, China. College of Life Science, Northeast Agriculture University, Harbin 150030, P.R. China.
Fang Qing-chang
Affiliation:
College of Life Science, Northeast Agriculture University, Harbin 150030, P.R. China.
Ren Liang
Affiliation:
College of Life Science, Northeast Agriculture University, Harbin 150030, P.R. China.
Ma Hong
Affiliation:
Institute of Animal Science, Heilongjiang Academy of Agriculture Science, Harbin 150086, P.R. China.
Wang Liang
Affiliation:
Institute of Animal Science, Heilongjiang Academy of Agriculture Science, Harbin 150086, P.R. China.
Guo Zhen-hua
Affiliation:
Institute of Animal Science, Heilongjiang Academy of Agriculture Science, Harbin 150086, P.R. China.
Li Zhong-qiu
Affiliation:
Institute of Animal Science, Heilongjiang Academy of Agriculture Science, Harbin 150086, P.R. China.
*
All correspondence to: Liu Di. Heilongjiang Academy of Agricultural Science; 368 Xue Fu Road, Harbin City 150086, China. Tel: +86 13845120192. e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

Transgenes integrated into mammalian cells are silenced rapidly. This phenomenon correlates with repressed chromatin structure marked by histone hypoacetylation. This study investigated the effect of trichostatin A (TSA; a histone-deacetylase inhibitor) on EGFP expression in transfected cells and embryonic development after somatic cell nuclear transfer (SCNT). Porcine adult fibroblasts were transfected with a pEGFP-C1 vector. Then transfected cells, donor cells for SCNT, were pretreated with TSA, with the untreated cells being used as the control. Expression of EGFP in donor cells and reconstructed embryos was detected when exposed to blue light. Results showed that the percentage of EGFP-positive cells significantly increased when the transfected cells were treated with TSA and the increased expression of EGFP was sustained to at least the morula stage. In addition, the cytotoxic effect of TSA on the transfected cells was dose dependent. In conclusion, TSA can rescue the silenced EGFP gene. Even after transferring the TSA-treated cells to enucleated recipient oocytes, TSA retained the ability to rescue a silenced EGFP gene. In addition, TSA had an impact on cell proliferation.

Keywords

EGFPHistone deacetylationPorcineSomatic cell nuclear transferTrichostatin A
Type
Research Article
Information
Zygote , Volume 19 , Issue 3 , August 2011 , pp. 237 - 243
Copyright
Copyright © Cambridge University Press 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Bhaumik, S.R., Smith, E. & Shilatifard, A. (2007). Covalent modifications of histones during development and disease pathogenesis. Nat. Struct. Mol. 14, 1008–16.Google Scholar
Choi, K.H., Basma, H., Singh, J. & Cheng, P.W. (2005). Activation of CMV promoter-controlled glycosyltransferase and beta-galactosidase glycogenes by butyrate, trichostatin A, and 5-aza-2′-deoxycytidine. Glycoconj. J. 22, 63–9.Google Scholar
Csordas, A. (1990). On the biological role of histone acetylation. Biochem. J. 265, 23–38.Google Scholar
Eden, S., Hashimshony, T., Keshet, I., Cedar, H. & Thorne, A.W. (1998). DNA methylation models histone acetylation. Nature 394, 842.CrossRefGoogle ScholarPubMed
Finnin, M.S., Donigian, J.R., Cohen, A., Richon, V.M., Rifkind, R.A., Marks, P.A., Breslow, R. & Pavletich, N.P. (1999). Structures of a histone deacetylase homologue bound to the TSA and SAHA inhibitors. Nature 401, 188–93.CrossRefGoogle Scholar
Grassi, G., Maccaroni, P., Meyer, R., Kaiser, H., D'Ambrosio, E., Pascale, E., Grassi, M., Kuhn, A., Di Nardo, P., Kandolf, R. & Küpper, J.-H. (2003). Inhibitors of DNA methylation and histone deacetylation activate cytomegalovirus promoter-controlled reporter gene expression in human glioblastoma cell line U87. Carcinogenesis 24, 1625–35.Google Scholar
Hu, J.F., Pham, J., Dey, I., Li, T., Vu, T.H. & Hoffman, A.R. (2000). Allele-specific histone acetylation accompanies genomic imprinting of the insulin-like growth factor II receptor gene. Endocrinology 141, 4428–4435.Google Scholar
Jenuwein, T. & Allis, C.D. (2001). Translating the histone code. Science 293, 1074–80.CrossRefGoogle ScholarPubMed
Kikuchi, H. & Fujimoto, D. (1973). Multiplicity of histone deacetylase from calf thymus. FEBS Lett. 29, 280–282.CrossRefGoogle ScholarPubMed
Krishnan, M., Park, J.M., Cao, F., Wang, D., Paulmurugan, R., Tseng, J.R., Gonzalgo, M.L., Gambhir, S.S. & Wu, J.C. (2006). Effects of epigenetic modulation on reporter gene expression: implications for stem cell imaging. FASEB J. 20, 106–8.Google Scholar
Löser, P., Jennings, G.S., Strauss, M. & Sandig, V. (1998). Reactivation of the previously silenced cytomegalovirus major immediate-early promoter in the mouse liver: involvement of NFkappaB. J. Virol. 72, 180–90.Google Scholar
Li, E. (2002). Chromatin modification and epigenetic reprogramming in mammalian development. Nat. Rev. Genet. 3, 662–73.CrossRefGoogle ScholarPubMed
Meier, J.L. (2001). Reactivation of the human cytomegalovirus major immediate-early regulatory region and viral replication in embryonal NTera2 cells: role of trichostatin A, retinoic acid, and deletion of the 21-base-pair repeats and modulator. J. Virol. 75, 1581–93.Google Scholar
Niemann, H., Rath, D. & Wrenzycki, C. (2003). Advances in biotechnology: new tools in future pig production for agriculture and biomedicine. Reprod. Domest. Anim. 38, 82–9.CrossRefGoogle ScholarPubMed
Ou, J.N., Torrisani, J., Unterberger, A., Provençal, N., Shikimi, K., Karimi, M., Tomas, J., Ekström, T.J. & Szyf, M. (2007). Histone deacetylase inhibitor trichostatin A induces global and gene-specific DNA demethylation in human cancer cell lines. Biochem. Pharmacol. 73, 1297–307.CrossRefGoogle ScholarPubMed
Prendergast, F.G. & Mann, K.G. (1978). Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskålea. Biochemistry 17, 3448–53.CrossRefGoogle ScholarPubMed
Schübeler, D., Lorincz, M.C., Cimbora, D.M., Telling, A., Feng, Y.-Q., Bouhassira, E.E. & Groudine, M. (2000). Genomic targeting of methylated DNA: influence of methylation on transcription, replication, chromatin structure, and histone acetylation. Mol. Cell. Biol. 20, 9103–12.Google Scholar
Tsien, R.Y. (1998). The green fluorescent protein. Annu. Rev. Biochem. 67, 509–44.Google Scholar
Verma, I.M. & Somia, N. (1997). Gene therapy—promises, problems and prospects. Nature 389, 239–42.CrossRefGoogle Scholar
Vogelstein, B. & Kinzler, K.W. (2004). Cancer genes and the pathways they control. Nat. Med. 10, 789–99.Google Scholar
Wee, G.., Shim, J.-J., Koo, D.-B., Chae, J.I., Lee, K.K. & Han, Y.M. (2007). Epigenetic alteration of the donor cells does not recapitulate the reprogramming of DNA methylation in cloned embryos. Reproduction 134, 781–7.Google Scholar
Yoshida, M., Kijima, M., Akita, M. & Beppu, T. (1990). Potent and specific inhibition of mammalian histone deacetylase both in vivo and in vitro by trichostatin A. J. Biol. Chem. 265, 17174–9.Google Scholar