Establishment of a model for axonal regeneration following mechanical transection in vitro

Abstract

Objective To establish a model fit for axonal regeneration research after its mechanical injury.Methods Cortical explants from mice were planted on culture dishes by microglass pipettes or routine glass pipettes.The cell body and dendrites in axonal area were detected by immunofluorescence and RT-PCR.Besides,purity of regenerated axons was also tested by immunofluorescence and RT-PCR after mechanical transection of axons under microscopy.Results Compared with explants planting by routine glass pipettes,in the outside 1/2 axons of explants planted by micro-glass pipettes,the immunofluorescence and RT-PCR showed negative nucleus and dendrites.In the regenerated axons following mechanical transection of explants planted by micro-glass pipette,the immunofluorescene and RT-PCR showed no regenerated axons nucleus mixed into the dendrites and nucleus.Conclusions Explants planted by micro-glass pipette obtains enough pure axons and regenerated axons.The establishment of models of axonal mechanical transection lays foundation for its molecular study after trauma. Key words: Axotomy; Regeneration; Model