title: Two-photon imaging of stroke onset in vivo reveals that NMDA-receptor independent ischemic depolarization is the major cause of rapid reversible damage to dendrites and spines.
abstract: We adapt a mouse global ischemia model to permit rapid induction of ischemia and reperfusion in conjunction with two-photon imaging to monitor the initial ionic, structural, and functional implications of brief interruptions of blood flow (6-8 min) in vivo. After only 2-3 min of global ischemia, a wide spread loss of mouse somatosensory cortex apical dendritic structure is initiated during the passage of a propagating wave (3.3 mm/min) of ischemic depolarization. Increases in intracellular calcium levels occurred during the wave of ischemic depolarization and were coincident with the loss of dendritic structure, but were not triggered by reperfusion. To assess the role of NMDA receptors, we locally applied the antagonist MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate] at concentrations sufficient to fully block local NMDA agonist-evoked changes in intracellular calcium levels in vivo. Changes in dendritic structure and intracellular calcium levels were independent of NMDA receptor activation. Local application of the non-NMDA glutamate receptor antagonist CNQX also failed to block ischemic depolarization or rapid changes in dendrite structure. Within 3-5 min of reperfusion, damage ceased and restoration of synaptic structure occurred over 10-60 min. In contrast to a reperfusion promoting damage, over this time scale, the majority of spines and dendrites regained their original structure during reperfusion. Intrinsic optical signal imaging of sensory evoked maps indicated that reversible alteration in dendritic structure during reperfusion was accompanied by restored functional maps. Our results identify glutamate receptor-independent ischemic depolarization as the major ionic event associated with disruption of synaptic structure during the first few minutes of ischemia in vivo.
authors.affiliation: Kinsmen Laboratory, Department of Psychiatry, Brain Research Center, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3. thmurphy@interchange.ubc.ca
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Murphy
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.forename: Kellen
authors.initials: K
authors.lastname: Betts
authors.forename: Richard
authors.initials: R
authors.lastname: Liu
mesh_terms: D000818:Animals; D001931:Brain Mapping / Q000379:methods*; D002118:Calcium / Q000378:metabolism; D003712:Dendrites / Q000473:pathology*; D049229:Dendritic Spines / Q000473:pathology; D004195:Disease Models, Animal; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004569:Electroencephalography; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020521:Stroke / Q000378:metabolism* / Q000473:pathology*; D013569:Synapses / Q000473:pathology
chemical_list: D018691:Excitatory Amino Acid Antagonists; D016194:Receptors, N-Methyl-D-Aspartate; D016291:Dizocilpine Maleate; D002118:Calcium