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Text
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URL Address
<a href="http://doi.org/10.1016/j.neuroscience.2007.02.026" target="_blank" rel="noreferrer noopener">http://doi.org/10.1016/j.neuroscience.2007.02.026</a>
Pages
773–783
Issue
2
Volume
146
Dublin Core
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Title
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On the use of retrograde tracers for identification of axon collaterals with multiple fluorescent retrograde tracers.
Publisher
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Neuroscience
Date
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2007
2007-05
Subject
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Amidines; Animals; Axons/metabolism/*physiology; Brain Mapping; Brain/*anatomy & histology; Coloring Agents/administration & dosage/metabolism; Dextrans/administration & dosage/metabolism; Female; Fluoresceins/administration & dosage/metabolism; Guinea Pigs; Indicators and Reagents/administration & dosage; Male; Neural Pathways/*anatomy & histology/metabolism
Creator
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Schofield B R; Schofield R M; Sorensen K A; Motts S D
Description
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A common method for identifying collateral projections is to inject different retrograde tracers into two targets and examine labeled cells for the presence of both tracers. Double-labeled cells are considered to have collateral projections to the two injection sites. This method is widely considered to underestimate the extent of collaterals. To test the efficiency of double-labeling, we mixed equal volumes of two tracers, injected them into one site in a guinea-pig brain, and counted the resulting labeled cells. Ideally, the tracers would have precisely overlapping injection sites and all labeled cells would contain both tracers. We tested several combinations of tracers: 1) Fast Blue and fluorescein dextran; 2) fluorescein dextran and FluoroGold; 3) fluorescein dextran and FluoroRuby; 4) FluoroGold and green beads; 5) FluoroGold and red beads; 6) FluoroRuby and green beads; and, 7) green beads and red beads. For each combination, a mixture was injected into the left inferior colliculus. After 1 week to allow for transport, labeled cells were counted in the right inferior colliculus and the left temporal cortex. For each mixture, the results were similar for the two areas. The percentage of cells that were double-labeled varied from 0% to 100%, depending on tracer combination. The highest efficiencies (\textgreater96%) were observed with red beads and green beads or with FluoroRuby and fluorescein dextran. The limited efficiency of other mixtures could be accounted for only in part by incomplete overlap of the two tracers at the injection site. The results indicate that the specific combination of tracers used to search for collateral projections can greatly affect the findings.
Identifier
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<a href="http://doi.org/10.1016/j.neuroscience.2007.02.026" target="_blank" rel="noreferrer noopener">10.1016/j.neuroscience.2007.02.026</a>
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Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
2007
Amidines
Animals
Axons/metabolism/*physiology
Brain Mapping
Brain/*anatomy & histology
Coloring Agents/administration & dosage/metabolism
Department of Anatomy & Neurobiology
Dextrans/administration & dosage/metabolism
Female
Fluoresceins/administration & dosage/metabolism
Guinea Pigs
Indicators and Reagents/administration & dosage
Male
Motts S D
NEOMED College of Medicine
Neural Pathways/*anatomy & histology/metabolism
Neuroscience
Schofield B R
Schofield R M
Sorensen K A