Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining

  • 37 Accesses

  • 46 Citations

Abstract

A relatively simple method has been developed to improve the resolution for measuring breaks produced in interphase chromosomes by X rays or other agents following the induction of premature chromosome condensation (PCC). Mitotic HeLa cells, which induce PCC when fused with interphase cells, were obtained from cultures grown for several generations in 5-bromodeoxyuridine (BrdU). These were fused to cells from low-passage confluent cultures of normal human fibroblasts and subsequently stained by a modified fluorescence-plus-Giemsa (FPG) technique. Following this protocol the prematurely condensed chromosomes stain intensely, whereas the mitotic chromosomes of the inducer cell(s), which are intermingled with them, stain very lightly. With this technique the interphase chromosomes and their fragments can be identified unequivocally, making scoring much easier and more accurate. The frequency of breaks produced in G1 phase AG1522 human fibroblasts immediately following X-ray doses of 58 and 117 rad was 3.68 and 7.38 per cell, respectively. Use of this technique should allow the detection of damage from ionizing radiation at doses lower than 10 rad.

This is a preview of subscription content, log in to check access.

References

  1. Ham RG, Puck TT (1962) Quantitative colonial growth of isolated mammalian cells. In: Colowick SP, Kaplan NO (eds) Methods in enzymology, Vol 5. Academic Press, New York, pp 90–119

  2. Hittelman WN, Pollard M (1982a) A comparison of DNA and chromosome repair kinetics after γ-irradaition. Rad Res 92:497–509

  3. Hittelman WN, Pollard M (1982b) Induction and repair of DNA and chromosome damage by neocarzinostatin in quiescent normal human fibroblasts. Cancer Res 42:4584–4590

  4. Hittelman WN, Rao PN (1974) Premature chromosome condensation, I. Visualization of X-ray induced chromosome damage in interphase cells. Mutat Res 23:251–258

  5. Johnson RT, Rao PN (1970) Mammalian cell fusion: Induction of premature chromosome condensation in interphase nuclei. Nature 226:717–722

  6. Johnson RT, Collins ARS, Waldren CA (1982) Prematurely condensed chromosomes and the analysis of DNA and chromosome lesions. In: Rao PN, Johnson RT, Sperling K (eds) Premature chromosome condensation. Academic Press, New York, pp 253–308

  7. Perry P, Wolff S (1974) New giemsa method for the differential staining of sister chromatids. Nature 251:156–158

  8. Rao PN, Wilson BA, Puck TT (1977) Premature chromosome condensation and cell cycle analysis. J Cell Physiol 91:131–142

  9. Waldren CA, Johnson RT (1974) Analysis of interphase chromosome damage by means of premature chromosome condensation after X- and ultraviolet-irradiation. Proc Natl Acad Sci 71:1137–1141

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cornforth, M.N., Bedford, J.S. High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining. Chromosoma 88, 315–318 (1983). https://doi.org/10.1007/BF00292909

Download citation

Keywords

  • HeLa Cell
  • Developmental Biology
  • Human Fibroblast
  • Mitotic Chromosome
  • Inducer Cell