The sequence of ultrastructural changes in cultured neurons after dendrite transection

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Abstract
Cultured mouse spinal neurons were fixed at three different intervals after dendrite amputation: within the first 15 min, at 2 h and at 24 h. Dendrites were amputated at lesion distance of either 50 μm (31% probability of cell survival) or 100 μm (53% probability of cell survival) from the edge of their perikarya. When fixed within 15 min, operated neurons showed a two-phase gradient of ultrastructural damage which spread from the transection site towards the perikaryon. At 2 h after dendrite amputation all neurons operated close to their perikarya were categorized as either viable, moribund or dead, based on their appearance with phase contrast microscopy. These categories of response to physical trauma corresponded to distinctly different ultrastructural changes. Moribund neurons were filled with membrane-bound vesicles which were derived from swollen mitochondria and grossly dilated cisternae of the smooth endoplasmic reticulum. The cytoplasm of dead neurons contained large clear areas and many condensed, dark mitochondria. Both moribund and dead neurons lacked cytoskeletal elements. All of these ultrastructural changes are hypothesized to be the result of an increase in the intracellular concentrations of free calcium. Although evidence of residual mitochondrial swelling was present in some surviving neurons at 24 h, the ultrastructure of others was comparable to that of control cells. Some surviving neurons had terminal swellings at the ends of the severed neurites which were very similar to retraction balls of transected axons after CNS trauma.