Document Type
Article
Publication Date
1982
DOI
10.1177/30.5.6176614
Publication Title
Journal of Histochemistry and Cytochemistry
Volume
30
Issue
5
Pages
425-435
Abstract
Eight methods for the electron microscopic demonstration of horseradish peroxidase (HRP) labeling have been compared in adjacent series of vibratome sections of mouse lumbar spinal cord. The tracer, a HRP-wheat germ agglutinin (WGA) conjugate, was injected into the gastrocnemius muscle complex. Following retrograde axonal transport to the lumbar motor neurons and transganglionic anterograde transport of the tracer to the dorsal horn, the HRP activity was demonstrated in eight series of adjacent sections of lumbar spinal cord using eight methods. These included procedures using tetramethylbenzidine (TMB), benzidine dihydrochloride (BDHC), o-tolidine, paraphenylenediamine-pyrocatechol (PPD-PC), and 4 methods using 3,3'-diaminobenzidine (DAB). All eight methods were able to demonstrate both retrograde labeling of motor neurons and transganglionic anterograde transport into the dorsal horn. However, there were differences in the appearance of the various reaction products under the electron microscope. In addition, differences in the distribution of the reaction products were observed by both light and electron microscopy. The largest distribution of reaction product was observed with TMB. BDHC and o-tolidine were next, followed by the DAB procedures and PPD-PC. The TMB, BDHC, and o-tolidine reaction products were all found to be suitable for electron microscopy. The TMB reaction product was electron dense and had a very distinctive crystalloid appearance that made identification of HRP-labeled neuronal profiles easy and unequivocal. Copyright © 1982 by The Histochemical Society, Inc.
Original Publication Citation
Carson, K. A., & Mesulam, M. M. (1982). Electron microscopic demonstration of neural connections using horseradish peroxidase: A comparison of the tetramethylbenzidine procedure with seven other histochemical methods. Journal of Histochemistry and Cytochemistry, 30(5), 425-435. doi:10.1177/30.5.6176614
Repository Citation
Carson, Keith A. and Mesulam, M-Marsel, "Electron Microscopic Demonstration of Neural Connections Using Horseradish Peroxidase: A Comparison of the Tetramethylbenzidine Procedure With Seven Other Histochemical Methods" (1982). Biological Sciences Faculty Publications. 357.
https://digitalcommons.odu.edu/biology_fac_pubs/357