Live cell imaging of heavy-ion-induced radiation responses by beamline microscopy

Jakob, B., Rudolph, J.H., Gueven, N., Lavin, M.E. and Taucher-Scholz, G. (2005) Live cell imaging of heavy-ion-induced radiation responses by beamline microscopy. Radiation Research, 163 6: 681-690. doi:10.1667/RR3374


Author Jakob, B.
Rudolph, J.H.
Gueven, N.
Lavin, M.E.
Taucher-Scholz, G.
Title Live cell imaging of heavy-ion-induced radiation responses by beamline microscopy
Journal name Radiation Research   Check publisher's open access policy
ISSN 0033-7587
Publication date 2005-06-01
Sub-type Article (original research)
DOI 10.1667/RR3374
Volume 163
Issue 6
Start page 681
End page 690
Total pages 10
Editor S. Rockwell
R. J. M. Fry
J. E. Moulder
et al.
Place of publication Oak Brook, U.S.A.
Publisher Radiation Research Society
Language eng
Subject C1
270100 Biochemistry and Cell Biology
730107 Inherited diseases (incl. gene therapy)
Abstract To study the dynamics of protein recruitment to DNA lesions, ion beams can be used to generate extremely localized DNA damage within restricted regions of the nuclei. This inhomogeneous spatial distribution of lesions can be visualized indirectly and rapidly in the form of radiation-induced foci using immunocytochemical detection or GFP-tagged DNA repair proteins. To analyze faster protein translocations and a possible contribution of radiation-induced chromatin movement in DNA damage recognition in live cells, we developed a remote-controlled system to obtain high-resolution fluorescence images of living cells during ion irradiation with a frame rate of the order of seconds. Using scratch replication labeling, only minor chromatin movement at sites of ion traversal was observed within the first few minutes of impact. Furthermore, time-lapse images of the GFP-coupled DNA repair protein aprataxin revealed accumulations within seconds at sites of ion hits, indicating a very fast recruitment to damaged sites. Repositioning of the irradiated cells after fixation allowed the comparison of live cell observation with immunocytochemical staining and retrospective etching of ion tracks. These results demonstrate that heavy-ion radiation-induced changes in sub-nuclear structures can be used to determine the kinetics of early protein recruitment in living cells and that the changes are not dependent on large-scale chromatin movement at short times postirradiation. © 2005 by Radiation Research Society.
Keyword Biology
Biophysics
Radiology, Nuclear Medicine & Medical Imaging
Double-strand Breaks
Nucleotide Excision-repair
Dna-damage
Living Cells
In-vivo
Chromatin
Recognition
Dynamics
Sites
Motion
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
2006 Higher Education Research Data Collection
School of Medicine Publications
 
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Created: Wed, 15 Aug 2007, 16:50:16 EST