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Journal of Neuroscience, Vol 16, 2740-2749, Copyright © 1996 by Society for Neuroscience
Intrinsic optical signals in rat neocortical slices measured with near- infrared dark-field microscopy reveal changes in extracellular space
K Holthoff and OW Witte
Neurologische Klinik der Heinrich Heine Universitat, Dusseldorf, Germany.
In the last decade, the measurement of activity-dependent intrinsic optical
signals (IOSs) in excitable tissues has become a useful tool for collecting
data about spatial patterns of information processing in mammalian brain
and spread of excitation. Although the extent of the IOS correlates well
with the extent of electrical excitation, its time course is much slower,
suggesting that it does not directly monitor the electrical activity. The
aim of this study was to investigate the mechanisms responsible for
generation of IOSs. Coronal neocortical brain slices of juvenile rats were
electrically stimulated at the border of layer VI and the white matter. The
induced columnar-shaped IOSs were recorded using dark-field video
microscopy. At corresponding locations, alterations in extracellular K+
concentration and extracellular space (ECS) volume were registered using
ion-selective microelectrodes. After stimulation, a transient increase of
extracellular K+ concentration up to 10 mM and a transient decrease of ECS
volume by approximately 4% could be observed. The comparison of the time
courses of these parameters yielded considerable differences between
extracellular K+ concentration increase and IOS, but obvious similarities
between alterations in ECS volume and IOS. To test the hypothesis that
changes in IOS reflect changes in ECS, but not extracellular K+
concentration, we recorded under conditions that are known to prevent
activity-induced changes in ECS, i.e., in low Cl- solutions and in the
presence of furosemide. Both treatments similarly decreased
stimulation-induced IOSs and alterations of ECS. However, the effect of
these treatments on changes of extracellular K+ was different and did not
correspond to the changes of IOS. We conclude that activity- dependent IOSs
in rat neocortical slices measured by near-infrared video microscopy reveal
changes in ECS. Furthermore, the pharmacological and ion substitutional
experiments make it likely that activity-induced IOSs are attributable to
cell swelling via a net KCI uptake and a concomitant water influx.
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