Fast optical recordings of membrane potential changes from dendrites of pyramidal neurons.
Understanding the biophysical properties of single neurons and how they process information is fundamental to understanding how the brain works. A technique that would allow recording of temporal and spatial dynamics of electrical activity in neuronal processes with adequate resolution would facilitate further research. Here, we report on the application of optical recording of membrane potential transients at many sites on neuronal processes of vertebrate neurons in brain slices using intracellular voltage-sensitive dyes. We obtained evidence that 1) loading the neurons with voltage-sensitive dye using patch electrodes is possible without contamination of the extracellular environment; 2) brain slices do not show any autofluorescence at the excitation/emission wavelengths used; 3) pharmacological effects of the dye were completely reversible; 4) the level of photodynamic damage already allows meaningful measurements and could be reduced further; 5) the sensitivity of the dye was comparable to that reported for invertebrate neurons; 6) the dye spread approximately 500 micron into distal processes within 2 h incubation period. This distance should increase with longer incubation; 7) the optically recorded action potential signals from basolateral dendrites (that are difficult or impossible to approach by patch electrodes) and apical dendrites show that both direct soma stimulation and synaptic stimulation triggered action potentials that originated near the soma. The spikes backpropagated into both basolateral dendrites and apical processes; the propagation was somewhat faster in the apical dendrites.
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Department of Cellular and Molecular Physiology, Yale University, School of Medicine, New Haven, Connecticut 06520, USA.
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Zecevic
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D000200:Action Potentials / Q000502:physiology; D000818:Animals; D016477:Artifacts; D004396:Coloring Agents; D003712:Dendrites / Q000502:physiology*; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000502:physiology; D055095:Optics and Photonics; D017966:Pyramidal Cells / Q000502:physiology*; D011833:Radiation Injuries, Experimental / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D013003:Somatosensory Cortex / Q000166:cytology / Q000473:pathology / Q000502:physiology / Q000528:radiation effects
D004396:Coloring Agents