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Combined Vital Dye Labelling and Catecholamine Histofluorescence of Transplanted Ciliary Ganglion Cells

Sechrist, John and Coulombe, James N. and Bronner-Fraser, Marianne (1989) Combined Vital Dye Labelling and Catecholamine Histofluorescence of Transplanted Ciliary Ganglion Cells. Journal of Neural Transplantation, 1 (3-4). pp. 113-128. ISSN 1352-237X. doi:10.1155/NP.1989.113.

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We have utilized the carbocyanine dye, Dil, to label suspensions of dissociated ciliary ganglion cells removed from 6 to 12 day old quail embryos. Some of the cells were injected into the trunk somites of 2.5 - 3 day old chick embryos along pathways where neural crest cells migrate to form sensory and sympathetic ganglia, aortic plexuses and the adrenal medulla; the remainder of the cells were cultured to check their viability and the persistence of the Dil label. Embryos were incubated for 1 - 8 days post-injection, fixed in 4% paraformaldehyde/0.25% glutaraldehyde and processed for cryostat sectioning. DH-labelled cells were readily identifiable in culture and in sections of embryos at all stages examined. Several cell types were identified, based on their morphology and soma size. These included cells with large cell bodies and bright Dil-labelling that appeared to be neurons and smaller, more weakly labelled cells that appeared non-neuronal. The latter presumably bad divided several times, accounting for their reduced levels of dye. Many of the Dil-labelled cells were found in and around neural crestderived sympathetic ganglia, aortic plexuses and adrenomedullary cords, but were rarely observed in dorsal root ganglia. The aldehyde fixative (Faglu mixture) used in this study reacts with catecholamines to form a bright reaction product in adrenergic cells including those in the sympathetic ganglia and the adrenal medulla. The catecholamine biproduct and the Dil in the same cell can easily be viewed with different fluorescent filter sets. A variable number of the DU-labelled cells in these adrenergic sites contained catecholamines. Cells derived from younger 6 day ciliary ganglion dissociates exhibited detectable catecholamine neurotransmitters earlier and more frequently than those derived from 8 day embryos. The presence of cells exhibiting both bright Dil and catecholamine fluorescence is consistent with previous indications that post-mitotic ciliary ganglion neurons can undergo pbenotypic conversion from cholinergic to adrenergic when transplanted to the trunk environment.

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Bronner-Fraser, Marianne0000-0003-4274-1862
Additional Information:© 1989 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. We thank Dr. Gabrielle Leblanc for a critical reading of the manuscript and Michael Artinger for his technical assistance. This work was supported by BNS 8809454.
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NSFBNS 8809454
Issue or Number:3-4
Record Number:CaltechAUTHORS:20160520-152504213
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Official Citation:Combined Vital Dye Labelling and Catecholamine Histofluorescence of Transplanted Ciliary Ganglion Cells John Sechrist, James N. Coulombe, and Marianne Bronner-Fraser Journal of Neural Transplantation Volume 1 (1989), Issue 3-4, Pages 113-128
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:67229
Deposited By: Ruth Sustaita
Deposited On:20 May 2016 23:02
Last Modified:11 Nov 2021 00:29

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