Mature oligodendrocyte (OL) translatome changes at various times after moderate contusive spinal cord injury at the thoracic T9 level
OL translatome was determined in female RiboTag:Plp1-CreERT2 mice (C57Bl6 background) at 2, 10, and, 42 days after moderate contusive T9 SCI. Ribotag expression was induced by daily tamoxifen injections (1 mg i.p. over 8 days). Three weeks after completion of the induction protocol moderate contusive SCI was performed at T9 level (IH imactor, 50 kdyn). Uninjured mice (naive) were used as controls. At the time of surgery, animal age was 10-12 weeks. After euthanasia, RNA was isolated from a 5 mm spinal cord segment spanning the injury epicenter. OL tranlsatome was immunopurified using anti-HA antibody and protein A/G coupled magnetic beads following the Ribotag protocol. Total spinal cord RNA and OL translatome RNA samples were analyzed by RNASeq. Each biological sample was prepared by pooling material from two mice (3 biological samples representing 6 animals were used for each timepoint). Overall design: Analyzing effects of spinal cord injury on mRNA levels in total spinal cord transcriptome and OL translatome