Summary
Lumbar sympathetic ganglia of 12-day-old chick embryos were cultured in organ cultures for 14 days with 1, 10 or 100 mg/l of hydrocortisone or without it. Catecholamines were demonstrated by the formaldehyde-induced fluorescence method. For electron microscopy, the cultures were fixed with glutaraldehyde and osmium tetroxide.
Two types of cells with catecholamine fluorescence were observed in the control cultures: (1) weakly fluorescent sympathetic neurons and sympathicoblasts with long nerve fibres, which were the most common cell type in the explant, and (2) brightly fluorescent cells with or without fluorescent processes, which were less common and were scattered in the explant.
Hydrocortisone caused a great increase in the number of the brightly fluorescent cells. With 10 mg/l of hydrocortisone the increase was about ten-fold as compared with the control cultures. There was no change in the morphology of the cells, nor could any change be observed in the fluorescence intensity by eye.
Electron microscopically the mature neurons were the most common cell type on the surface of the culture, while more immature sympathicoblasts were seen in the deeper layers. Cells were also found which contained large numbers of catecholamine-storing granular vesicles 105–275 nm in diameter. These cells were infrequent. They had round vesicular nuclei and resembled also in other respects sympathicoblasts or young nerve cells. One such cell was found in mitotic division by electŕon microscopy.
Hydrocortisone caused a marked increase in the number of these granule-containing cells and their processes. Cells which could have been classified as the small intensely fluorescent cells of the mammalian ganglion type or their electron microscopic equivalent, the granule-containing cells were found neither in the control cultures nor in the hydrocortisone-containing cultures.
It is concluded that most brightly fluorescent cells in cultured sympathetic ganglia of the chick are nerve cells or sympathicoblasts rich in amine-storing granular vesicles.
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The authors are grateful to Dr. Margaret Murray for a copy of an unpublished manuscript and for stimulating discussions with one of the authors (O.E.), when he was a Fogarty Scholar-in-Residence in N.I.H., Bethesda, Md., U.S.A.
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Hervonen, H., Eränkö, O. Fluorescence histochemical and electron microscopical observations on sympathetic ganglia of the chick embryo cultured with and without hydrocortisone. Cell Tissue Res. 156, 145–166 (1975). https://doi.org/10.1007/BF00221799
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DOI: https://doi.org/10.1007/BF00221799