Tracing specific transmitter pathways in the rat CNS: combination of [3H]serotonin retrograde labelling with immunocytochemical detection of endogenous transmitters.

Journal of Neuroscience Methods
S AranedaAndré Calas

Abstract

Selective retrograde labelling with [3H]serotonin ([3H]5-HT) can be used to identify serotonergic cell bodies after specific [3H]5-HT uptake by the corresponding nerve terminals. In the present study, we demonstrate that autoradiography of this [3H]5-HT radiolabelling can be combined with immunocytochemical detection of endogenous serotonin, GABA or substance P on the same tissue section. The midbrain raphe serotonergic projections to the olfactory bulb and the spinal projections of medullary serotonergic nuclei were investigated. The specificity of retrograde labelling with [3H]5-HT was confirmed by immunoreactivity of the radiolabelled cells for serotonin, using an antiserum specific for formaldehyde-fixed serotonin. After spinal injections of [3H]5-HT, many retrogradely labelled cells in the medullary raphe were immunopositive for substance P, and a few for GABA. These results are in agreement with the available information on the co-existence of putative transmitters in the spinal projections of caudal raphe neurons. Therefore, autoradiography of [3H]5-HT retrograde labelling combined with immunocytochemistry offers a possibility to test the specificity of transmitter-selective retrograde labelling, to identify transmitter-...Continue Reading

References

Mar 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·G W Kreutzberg
Jun 1, 1980·The Journal of Comparative Neurology·P Streit
Mar 1, 1983·Journal of Neurochemistry·A Oblin, B Zivkovic
Jun 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·P SeguelaM Le Moal

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Citations

Jul 16, 2002·Hearing Research·Laura M HurleyGeorge D Pollak
Mar 10, 2000·Journal of Chemical Neuroanatomy·D Vergé, A Calas
Feb 24, 2010·Annales pharmaceutiques françaises·A Calas

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