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Retrograde horseradish peroxidase tracing combined with localization of choline acetyltransferase immunoreactivity.
- Source :
-
The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society [J Histochem Cytochem] 1984 Apr; Vol. 32 (4), pp. 439-43. - Publication Year :
- 1984
-
Abstract
- Localization of choline acetyltransferase (ChAT) immunoreactivity in rodent brain (AI Levey, DM Armstrong, SF Atweh, RD Terry, BH Wainer: J Neurosci 3 1, 1983) with a monoclonal antibody (Ab8) has been previously reported. Now a procedure for combining ChAT immunohistochemistry with retrograde tracing for the purpose of mapping cholinergic pathways is presented. Rats were injected with horseradish peroxidase-wheat germ agglutinin in the tongue and cerebral cortex. Sections from their perfusion-fixed brains were reacted with 3,3'-diaminobenzidine (DAB)/H2O2/cobalt acetate followed by ChAT localization with monoclonal antibody Ab8 using the peroxidase-antiperoxidase method and visualization using DAB/H2O2. Double-labeled cells were visualized with black punctate staining (retrograde tracer) on a diffuse brown cytoplasmic background (ChAT immunoreactivity) in the hypoglossal nucleus and ventral telencephalon (substantia innominata-nucleus basalis). Conditions of fixation, histochemistry, and immunohistochemistry that contribute to optimal resolution for this procedure are discussed.
Details
- Language :
- English
- ISSN :
- 0022-1554
- Volume :
- 32
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
- Publication Type :
- Academic Journal
- Accession number :
- 6368680
- Full Text :
- https://doi.org/10.1177/32.4.6368680