Pathway tracing using biotinylated dextran amines

Anton Reiner, C. Leonardus Veenman, Loreta Medina, Yun Jiao, Nobel Del Mar, Marcia Honig

Research output: Contribution to journalArticle

263 Citations (Scopus)

Abstract

Biotinylated dextran amines (BDA) are highly sensitive tools for anterograde and retrograde pathway tracing studies of the nervous system. BDA can be reliably delivered into the nervous system by iontophoretic or pressure injection and visualized with an avidin-biotinylated HRP (ABC) procedure, followed by a standard or metal-enhanced diaminobenzidine (DAB) reaction. High molecular weight BDA (10 k) yields sensitive and exquisitely detailed labeling of axons and terminals, while low molecular weight BDA (3 k) yields sensitive and detailed retrograde labeling of neuronal cell bodies. The detail of neuronal cell body labeling can be Golgi-like. BDA tolerates EM fixation and processing well and can, therefore, be readily used in ultrastructural studies. Additionally, BDA can be combined with other anterograde or retrograde tracers (e.g. PHA-L or cholera toxin B fragment) and visualized either by multi-color DAB multiple-labeling - if permanent labels are desired, or by using multiple simultaneous immunofluorescence - if fluorescence viewing is desired. In the same manner, BDA pathway tracing and neurotransmitter immunolabeling can be combined. Note that BDA pathway tracing can also be combined with anterograde or retrograde labeling with fluorescent dextran amines, if one wishes to exclusively use tracers with the favorable transport properties and sensitivities of dextran amines. In this case, the BDA can be visualized together with the fluorescent dextran amines using fluorescence labeling for the BDA, or the fluorescent dextran amines can be visualized together with the BDA by multicolor DAB labeling via immunolabeling of the fluorescent dextran amines using anti-fluorophore antisera. BDA is, thus, a flexible and valuable pathway tracing tool that has gained widespread popularity in recent years. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)23-37
Number of pages15
JournalJournal of Neuroscience Methods
Volume103
Issue number1
DOIs
StatePublished - Nov 15 2000

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Dextrans
Amines
Nervous System
biotinylated dextran amine
Molecular Weight
Fluorescence
Cholera Toxin
Presynaptic Terminals
Fluorescent Antibody Technique
Neurotransmitter Agents
Immune Sera
Color
Metals
Pressure
Injections
Cell Body

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Pathway tracing using biotinylated dextran amines. / Reiner, Anton; Veenman, C. Leonardus; Medina, Loreta; Jiao, Yun; Del Mar, Nobel; Honig, Marcia.

In: Journal of Neuroscience Methods, Vol. 103, No. 1, 15.11.2000, p. 23-37.

Research output: Contribution to journalArticle

Reiner, Anton ; Veenman, C. Leonardus ; Medina, Loreta ; Jiao, Yun ; Del Mar, Nobel ; Honig, Marcia. / Pathway tracing using biotinylated dextran amines. In: Journal of Neuroscience Methods. 2000 ; Vol. 103, No. 1. pp. 23-37.
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