Figure 6.

Mechanical injury of the spinal cord neurons cultured in a monolayer network. (A–C) Control. (D—F) Cultured with CM101 (0.3 μg/ml) 48 hr before injury and for the duration of the experiment. The patch–clamp pipette (X) was used to produce mechanical crush injury to single neurons. After obtaining control image (A), mild compression was applied to axons (arrowhead) to produce cell damage. Axon and soma swelled within 1–3 min (B), and axon autoamputation and autolysis was observed in 95% of neurons within 5–30 min. Within 1–4 hr, swelling was reduced (C) in the remaining injured untreated neurons. CM101 treatment (D) prevented cell death (95% survival) and axonal amputation (two injured neurons shown: arrowhead in E); however, CM101 did not protect axons from swelling (E), which occurred within 2–3 min after crush injury and reversed within 6 hr (F). Z, intracellular microelectrode used to record from a single neuron. (Bar = 50 μm.)