Ultrastructural damage and neuritic beading in cold-stressed spinal neurons with comparisons to NMDA and A23187 toxicity.
While exposure of cultured spinal neurons to mild hypothermia provides some protection from physical trauma (dendrotomy), profound cooling (< 17 degrees C) causes unrelated neuronal injury and death, which can be prevented by treatment with NMDA receptor antagonists. To investigate the mechanism of hypothermic neuronal injury we examined the ultrastructure of cultured spinal neurons after 2 h of cooling to 17 degrees C or 10 degrees C, with or without the presence of the NMDA receptor antagonist D-2-amino-5-phosphonovalerate, and with or without rewarming to 37 degrees C. These groups were compared to cultures exposed to NMDA or to the calcium ionophore A23187. Patterns of ultrastructural change, involving cytoskeletal disruption, mitochondrial abnormalities and vacuolization of the cytoplasm, suggest a common mechanism of injury in all treatment groups, involving an elevation of intracellular calcium. Some neurons exposed to hypothermia, NMDA or ionophore developed beaded dendrites. Microtubules were fragmented in varicosities but not in the intervening constrictions; other organelles were largely excluded from the constrictions. Varicosities may form when organelles and cytoplasm accumulate as the result of disruption of transport and membrane stabilizing proteins by proteases activated by calcium influx via NMDA mediated channels. The periodic nature of the swellings may reflect inherently discontinuous distribution of molecular subunits of the cytoskeleton.
Emery
D G
DG
Department of Zoology and Genetics, Iowa State University, Ames 50011-3223, USA.
Lucas
J H
JH
D015763:2-Amino-5-phosphonovalerate / Q000494:pharmacology; D000818:Animals; D000001:Calcimycin / Q000633:toxicity; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D003080:Cold Temperature / Q000009:adverse effects*; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D007476:Ionophores / Q000633:toxicity; D051379:Mice; D008854:Microscopy, Electron; D016202:N-Methylaspartate / Q000633:toxicity; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure*; D009474:Neurons / Q000187:drug effects / Q000648:ultrastructure*; D016194:Receptors, N-Methyl-D-Aspartate / Q000819:agonists; D013312:Stress, Physiological / Q000139:chemically induced / Q000473:pathology*; D014947:Wounds and Injuries / Q000473:pathology
D018690:Excitatory Amino Acid Agonists; D018691:Excitatory Amino Acid Antagonists; D007476:Ionophores; D016194:Receptors, N-Methyl-D-Aspartate; D000001:Calcimycin; D016202:N-Methylaspartate; D015763:2-Amino-5-phosphonovalerate; D002118:Calcium