Protection of the axonal cytoskeleton in anoxic optic nerve by decreased extracellular calcium.
Since CNS white matter tracts contain axons, oligodendrocytes and astrocytes but not synapses, it is likely that anoxic injury of white matter is mediated by cellular mechanisms that do not involve synapses. In order to test the hypothesis, that anoxic injury of white matter is mediated by an influx of Ca2+ into the intracellular compartment of axons, we compared the ultrastructure of axons in rat optic nerve exposed to 60 min of anoxia in artificial cerebrospinal fluid (aCSF) containing normal (2 mM) Ca2+, and in aCSF containing zero-Ca2+ together with 5 mM EGTA. Optic nerves fixed at the end of 60 min of anoxia in 2 mM Ca2+ exhibit extensive ultrastructural alterations including disruption of microtubules and neurofilaments within the axonal cytoskeleton, development of membranous profiles and empty spaces between the axon and the ensheathing myelin, and swelling of mitochondria with loss of cristae. Bathing the nerves in zero-Ca2+ aCSF during anoxia protected the axons from cytoskeletal changes; after 60 min of anoxia, optic nerve axons retained normal-appearing microtubules and neurofilaments. Membranous profiles were rare, and empty spaces between axons and myelin did not develop in anoxic optic nerves bathed in zero-Ca2+ aCSF. Disorganization of cristae in axonal mitochondria was observed in anoxic optic nerves even when Ca2+ was omitted from the medium. Because Ca(2+)-mediated injury is known to disrupt the axonal cytoskeleton, these results support the hypothesis that anoxia triggers an abnormal influx of Ca2+ into myelinated axons in CNS white matter.
Waxman
S G
SG
Department of Neurology, Yale School of Medicine, New Haven, CT 06510.
Black
J A
JA
Ransom
B R
BR
Stys
P K
PK
D000818:Animals; D001369:Axons / Q000648:ultrastructure*; D002118:Calcium / Q000378:metabolism / Q000502:physiology*; D003599:Cytoskeleton / Q000648:ultrastructure*; D002534:Hypoxia, Brain / Q000473:pathology*; D066298:In Vitro Techniques; D007382:Intermediate Filaments / Q000648:ultrastructure; D008870:Microtubules / Q000648:ultrastructure; D009413:Nerve Fibers, Myelinated / Q000378:metabolism; D009900:Optic Nerve / Q000648:ultrastructure*; D010477:Perfusion; D011901:Ranvier's Nodes / Q000648:ultrastructure; D051381:Rats
D002118:Calcium