| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1687195 | Nuclear Instruments and Methods in Physics Research Section B: Beam Interactions with Materials and Atoms | 2006 | 6 Pages |
Ion beams offer the possibility to generate strictly localized DNA lesions within subregions of a cell nucleus. The distribution of the ion-induced damage can be indirectly visualized by immunocytochemical detection of repair-related proteins as radiation-induced foci. The proteins analyzed here were the double-strand break marker γ-H2AX, the excision repair and replication protein PCNA and the cell cycle regulator CDKN1A. A newly developed adjustable sample holder is now used to apply an irradiation geometry characterized by a small angle between the plane of the cellular monolayer and the incoming ion beam. This allows the spatial analysis of protein accumulations along ion trajectories, revealing an unexpected clustering after irradiation with low-energy zinc ions. The patterns of protein aggregation observed show considerable intrinsic variability, but similar patterns of protein clustering were obtained for functionally different proteins irrespective of the type of ion beam applied, confirming previous observations for lower and higher LET beams. Foci sizes within ion tracks were found to be larger for γ-H2AX foci in comparison to CDKN1A foci, in agreement with the known histone H2AX phosphorylation response. The results suggest that not the pattern of dose deposition but the underlying chromatin structure determines the distribution of protein clusters along tracks. Therefore, the requirement of time-lapse studies using live cells is emphasized for future studies on chromatin movement as a potential component of the DNA damage response.
