Live Cell Imaging of Heavy-Ion-Induced Radiation Responses by Beamline Microscopy
| Autor: | G. Taucher-Scholz, Burkhard Jakob, Nuri Gueven, Martin F. Lavin, J. H. Rudolph |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
DNA damage
DNA repair Cell Culture Techniques Biophysics Biology Radiation Dosage chemistry.chemical_compound Live cell imaging Image Interpretation Computer-Assisted Microscopy Humans Heavy Ions Linear Energy Transfer Radiology Nuclear Medicine and imaging Irradiation Microscopy Video Radiation Nuclear Proteins Dose-Response Relationship Radiation DNA Equipment Design Robotics Molecular biology Chromatin DNA-Binding Proteins Equipment Failure Analysis Protein Transport chemistry DNA Damage HeLa Cells Nucleotide excision repair |
| Zdroj: | Radiation Research. 163:681-690 |
| ISSN: | 1938-5404 0033-7587 |
| DOI: | 10.1667/rr3374 |
| Popis: | 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 subnuclear 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. |
| Databáze: | OpenAIRE |
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