Assessing Cell Cycle Progression of Neural Stem and Progenitor Cells in the Mouse Developing Brain after Genotoxic Stress

• DOI: 10.3791/51209-v
• Publication Date: 2021-11-19T14:56:35Z
• Authors: Olivier Etienne; Amandine Bery; Telma Roque; Chantal Desmaze; François D. Boussin
• Keywords: 0303 health sciences; Stem Cells; Cell Cycle; Brain; Deoxyuridine; G1 Phase Cell Cycle Checkpoints; Mice; 03 medical and health sciences; Bromodeoxyuridine; Microscopy, Fluorescence; Neural Stem Cells; Pregnancy; S Phase Cell Cycle Checkpoints; Animals; Female; [SDV.BC] Life Sciences [q-bio]/Cellular Biology; Whole-Body Irradiation; DNA Damage

Neurons of the cerebral cortex are generated during brain development from different types neural stem and progenitor cells (NSPC), which form a pseudostratified epithelium lining lateral ventricles embryonic brain. Genotoxic stresses, such as ionizing radiation, have highly deleterious effects on developing related to high sensitivity NSPC. Elucidation cellular molecular mechanisms involved depends characterization DNA damage response these particular cells, requires an accurate method determine NSPC progression through cell cycle in damaged tissue. Here is shown based successive intraperitoneal injections EdU BrdU pregnant mice further detection two thymidine analogues coronal sections both incorporated replicating S phase detected by techniques (azide or specific antibody, respectively), facilitate their simultaneous detection. staining then determined for each nucleus function its distance ventricular margin standard region dorsal telencephalon. Thus this dual labeling technique allows distinguishing that progressed those activated checkpoint leading arrest damage. An example experiment presented, was injected before irradiation immediately after analyzes performed within 4 hr following irradiation. This protocol provides analysis acute at they been irradiated. easily transposable many other systems order impact treatment living tissues.