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Comparative Study
. 1994 Nov;26(11):856-62.
doi: 10.1007/BF00162931.

Retrograde neuronal tracing with cholera toxin B subunit: comparison of three different visualization methods

P J Dederen  1 , A A Gribnau, M H Curfs
Affiliations
Comparative Study

Retrograde neuronal tracing with cholera toxin B subunit: comparison of three different visualization methods

P J Dederen et al. Histochem J. 1994 Nov.

Abstract

In this report a comparison is made of three different visualization methods of rat cervical motoneurons retrogradely labelled with cholera toxin B subunit (CTb). CTb conjugates such as CTb-HRP and CTb-FITC or CTb-TRITC, which can be visualized after histochemical detection and by fluorescence microscopy, respectively. The following results were obtained. (1) Immunochemical detection of CTb with peroxidase and DAB-Ni incubation provides the best labelling of the cell bodies and their processes, whereas immunochemical detection with FITC produces less effective labelling of the dendrites. (2) Histochemical visualization of CTb-HRP conjugate gives results similar to those of CTb immunochemistry but produces a much more granular appearance of the label, which may affect the identification of distal dendrites. In addition, direct electron-microscopic analysis of labelled structures can be achieved. (3) CTb-FITC and CTb-TRITC visualization permit double-labelling experiments but the labelled cells exhibit fluorescence only in their somata and proximal dendrites. (4) Factors other than labelling intensity, e.g. double-labelling, preservation of the label, compatibility with other techniques and even economic reasons must be taken into consideration when a selection of visualization methods is to be made.

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