Neuron labeling by extracellular delivery of horseradish peroxidase in vivo: a method for studying the local circuitry of projection and interneurons at physiologically characterized sites

An anatomical method is described that yields individual neurons with continuously labeled dendrites and axons following the extracellular deposition of horseradish peroxidase (HRP) at neurophysiological recording sites in vivo. The method is a logical evolution of previous methods for iontophoretic...

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Bibliographic Details
Published in:Journal of neuroscience methods 1995-03, Vol.57 (1), p.81-91
Main Authors: Benson, Thane E., Voigt, Herbert F.
Format: Article
Language:English
Subjects:
Animals
Axon
Axons - physiology
Biological and medical sciences
Cochlea - cytology
Cochlea - physiology
Cochlear nucleus
Dendrite
Dendrites - physiology
Extracellular Space - physiology
Fundamental and applied biological sciences. Psychology
General aspects. Models. Methods
Gerbillinae
Glia
Horseradish peroxidase
Horseradish Peroxidase - administration & dosage
In vivo labeling
Interneurons - physiology
Iontophoresis
Local circuit
Microelectrodes
Microscopy, Electron
Neurophysiology
Vertebrates: nervous system and sense organs
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Summary:An anatomical method is described that yields individual neurons with continuously labeled dendrites and axons following the extracellular deposition of horseradish peroxidase (HRP) at neurophysiological recording sites in vivo. The method is a logical evolution of previous methods for iontophoretic delivery of HRP: Parameters critical to the ultimate concentration of HRP at the labeling site are reduced by an order of magnitude relative to standard practice. In successful cases one neuron or two in the immediate vicinity (50 μm) of recording sites is/are labeled. Labeling of other processes traversing the injection site, if any, is subliminal at highest light microscopic magnification. Due to the labeling of so few cells and the absence of other labeled processes, dendritic trees and local axonal arbors can be reconstructed without ambiguity. In addition to recovering neurons at sites characterized with physiological (e.g., sensory) stimuli, the method offers the further advantage of being fully compatible with subsequent electron microscopy. Both large (> 20 μm) and small (≈ 8 μm) neuron types and glia have been labeled.
ISSN:0165-0270
1872-678X
DOI:10.1016/0165-0270(94)00131-Y