Mechanostress induces DNA damage in migrating neurons during brain development
Newborn neurons in the developing cerebellar cortex undergo DNA double-strand breaks (DSBs) due to mechanostress during passage through narrow interstitial spaces. The purpose of this study was to map the DSB sites and to profile transcriptional changes in postmigratory neurons defective in the DSB repair pathway. Overall design: Dissociated cerebellar granule neurons subjected to confined migration (transwell assay) or control conditions (standard culture dishes) were harvested 8 hours after seeding for End-seq (with or without etoposide treatment), ChIP-seq, RNA-seq, or ATAC-seq. For tissue-based End-seq and ChIP-seq, cerebella from P15 control (Neurod1-Cre-negative ; Lig4 flox/flox) and Lig4 ND1-Cre (Neurod1-Cre-positive ; Lig4 flox/flox) mice were dissected, and nuclei were isolated either for direct lysis (End-seq) or crosslinking (ChIP). RNA-seq on cerebella from 2-month-old control and Lig4 ND1-Cre mice was performed by extracting total RNA from the tissue directly.