5/1/2023 0 Comments Geneious prime student account![]() ![]() Pdgfrα + :tdT + :DTA − littermates administered TAM and infused with vehicle alone served as non-ablated controls. Starting 4 days post-TAM, mice received a 6-day infusion of AraC before being sacrificed. TAM was administered to 8-week-old Pdgfrα + :tdT + :DTA + mice to induce expression of both tdTomato and DTA from the Ai14 tdTomato and Sox10-DTA recombined alleles, respectively ( Figure 2A ). To further explore the extent of OPC ablation, we generated Pdgfrα-CreER T2+/− :Ai14-tdTomato +/− :Sox10-DTA +/− mice (hereafter denoted Pdgfrα + :tdT + :DTA +), to enable simultaneous genetic fate-mapping and ablation of OPCs. ![]() Intracisternal infusion of AraC following TAM prevented rapid OPC regeneration However, residual non-recombined OPCs exhibited a robust proliferative response to OPC ablation, resulting in restoration of OPCs to similar or higher density than non-ablated controls within 8–10 days post-TAM. Together, these data demonstrate that the vast majority of OPCs in Pdgfrα + :DTA + mice were depleted after TAM administration. Supporting this idea, not all SOX10 + oligodendroglia in Pdgfrα + :DTA + brains examined 4 days post-TAM expressed GFP ( Figure S1A). Some EdU + OPCs did not express GFP ( Figure S1D), most likely reflecting low transcriptional activity of the Sox10 promoter that directs GFP expression. ![]() Most OPCs present 4 days post-TAM expressed GFP ( Figure S1D), suggesting that surviving OPCs were principally those in which the Sox10-DTA allele had not recombined. Indeed, most OPCs present after 10 days were newly generated, as demonstrated by the significant proportion of PDGFRA + cells incorporating 5-ethynyl-2′-deoxyuridine (EdU) provided continuously in the drinking water after TAM delivery ( Figures S1C and S1D). The marked increase in OPC density observed 10 days post-TAM suggests that OPCs exhibit robust proliferation following acute ablation. Although Pdgfrα + :DTA + mice exhibited marked OPC depletion 4 days post-TAM, OPC density in the corpus callosum returned to control levels by day 8 post-TAM and increased further over the subsequent 2 days ( Figures 1E–1G). ![]()
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |