Advertisement

Chinese Science Bulletin

, Volume 44, Issue 12, pp 1101–1106 | Cite as

Apoptosis in suspension culture of tobacco cells induced by heat shock

  • Haoming Chen
  • Changhui Yan
  • Xiaofang Jiang
  • Yaoren Dai
Notes

Abstract

The induction of apoptosis in suspension culture of tobacco cells by heat shock is reported for the first time. Heat treatment (48° for 4 h) of tobacco cells led to the appearance of typical hallmarks of apoptosis. It was demonstrated by DNA laddering analysis that the cells treated with heat shock at 48°C for 4 h had a serious degradation of nuclear DNA into multi-nucleosomal sizes, suggesting that heat shock activated endogenous nuclease which led to DNA cleavage at the linkage sites between the nucleosomes, but ladders were very faint for DNA from2 and 9 h heat-treated cells. The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) detection also showed that most of these treated cells (48°C for4 h) displayed positive reactions, indicating a serious DNA 3′-OH cleavage in their nuclei. Moreover, some other cytological changes in apoptotic cells, such as cell shrinkage, chromatin aggregation, nucleus collapse, have also been observed by 4′, 6′-diamidino-2-phenyl-indole (DAPI) staining.

Keywords

apoptosis DNA ladder TUNEL Nicotiana tabacum 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Kerr, J. F. R., Wylllie, A. H., Currie, A. H., Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics,Bri. J. Cancer., 1972, 26: 239.Google Scholar
  2. 2.
    McCabe, P. F., Levine, A., Meijer, P. J.et al., A programmed cell death pathway activated in carrot cells cultured at low density,Plant J., 1997, 12(2): 267.CrossRefGoogle Scholar
  3. 3.
    Jiang, X. F., Zhu, H. Z., Zhou, J.et al., Application of comet assay in plant protoplast apoptosis detection,Acta Botanica Sinica (in Chinese), 1998, 40(10): 928.Google Scholar
  4. 4.
    Poccia, F., Peselli, P., Vendetti, S.et al., Heat shock protein expression on the membrane of T-cell undergoing apoptosis,Immnunology, 1996, 88: 6.CrossRefGoogle Scholar
  5. 5.
    Li, W. X., Chen, C. H., Ling, C. C.et al., Apoptosis In heat induced cell killing the protective role of hsp-70 and the sensitization effect of the c-myc gene,Radh Res., 1996, 324.Google Scholar
  6. 6.
    Zhu, W. G., Aramaki, R., Cai, Y.et al., Promotion of heat-induced apoptosis in Fm3A cells by protease inhibitors,Biochem. Bioph. Res. Co., 1996, 225(3): 924.CrossRefGoogle Scholar
  7. 7.
    Vogel, P., Dux, E., Wiessner, C., Evidence of apoptosis in primary neuronal cultures after heat-shock.,Brain Res., 1997, 764(1-2): 205.PubMedCrossRefGoogle Scholar
  8. 8.
    Wang, H., Juan, L., Richard,. M. B.et al., Apoptosis: a functional paradigm for programmed plant cell death induced by a host-selective phytotoxin and invoked during development,Plant Cell. 1996, 8: 375.PubMedCrossRefGoogle Scholar
  9. 9.
    Mitter, R., Lam, E., Identification, characterization, and purification of a tobacco endonuclease activity induced on hypersensitive response cell death,Plant Cell, 1995, 7: 1951.CrossRefGoogle Scholar
  10. 10.
    Harm, H. K., Thermotoleranee in mammalian cells protein denaturation and aggregation, and stress proteins,J. Cell Sci., 1993, 104: 11.Google Scholar
  11. 11.
    Mosser, D. D., Caron, A. W., Bourget, L.et al., Role of the human hat-shock-protein Hsp70 in protection against stressinduced apoptosis,Mol. Cell Biol., 1997, 17(9): 5317.PubMedGoogle Scholar
  12. 12.
    Mehlen, P., Mehlen, A., Godet, J.et al., Hap27 as a switch between differentiation and apoptosis in murine embryonic stem-cells,J. Biol. Chem., 1997, 272(50): 31657.PubMedCrossRefGoogle Scholar

Copyright information

© Science in China Press 1999

Authors and Affiliations

  • Haoming Chen
    • 1
  • Changhui Yan
    • 1
  • Xiaofang Jiang
    • 1
  • Yaoren Dai
    • 1
  1. 1.Department of BiologyTsinghua UniversityBeijingChina

Personalised recommendations