Cell death in motor neurons of cultured spinal cord slices in adult mouse

Spinal cord slices culturing from adult mammals could be considered as a suitable in-vitro model for evaluating cellular viability, spinal cord injury and cell death mechanisms. In present study, determining of cell death in motor neurons of cultured spinal cord slices in adult mouse was done. In a experimental- laboratory study, thoracic regions of spinal cords from 4 Balb/c mice were cut into 400-µm slices using tissue chopper and incubated in a Co2 incubator at 37˚C for different periods of time. Freshly prepared slices (0h) and cultured slices were fixed and sectioned using cryostat. To study morphological and biochemical features of cell death, fluorescent staining, TUNEL method and agarose gel electrophoresis were used. In freshly prepared slices of motor neurons showed no apoptotic changes. While, 6, 12 and 24h after culturing, this neurons displayed morphological features of apoptosis including cell shrinkage as well as nuclear and chromatin condensation. Also, 6 and 12h after culturing were TUNEL positive. In addition, extracted DNA from cultured slices for 24h were indicated the nucleosomal DNA fragmentation on agarose gel electrophoresis. Results were showed the occurrence of apoptosis in motor neurons of cultured adult mouse spinal cord slices.