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Growth characteristics of postnatal rat adrenal medulla in culture

A study correlating phase contrast, microcinematographic, histochemical, and electron microscopical observations

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Summary

Explants and enzyme-dispersed cells of adrenal medulla from 10–12 day old rats were studied in culture for up to 3 weeks. Adrenomedullary chromaffin cells, nerve cells and satellite cells were clearly discernible.

The nerve cells were few in number and did not show catecholaminespecific fluorescence.

Chromaffin cells stored catecholamines, as judged by the Falck and Hillarp method, in varying amounts decreasing with age of the cultures and the distance from the explants. Exocytosis profiles observed with the electron microscope suggested that cultured chromaffin cells also released catecholamines. Moreover, the cells formed processes and frequently migrated into the outgrowth. After 6 days in culture, the great majority of chromaffin cells stored noradrenaline as revealed by electron microscopy with few adrenaline-storing cells being visible. Granular vesicles (∼ 80–240 nm in diameter) with cores of different electron densities were occasionally present in the same cell suggesting the occurrence of mixtures of primary and secondary amines. Apart from “chromaffin” granules, small clear and densecored vesicles (∼ 40–60 nm) were found both in the somata and cell processes. Chromaffin cells and their processes were often closely apposed and occasionally formed specialized attachment zones. As a whole, chromaffin cells in culture resembled small granule-containing cells in sympathetic ganglia.

0.5 mM dbcAMP prevented dedifferentiation of chromaffin cells as judged by the lack of processes, the size and amount of “chromaffin” granules and the high number of adrenaline-storing cells present after 6 days in culture.

NGF caused a striking increase in the number of axons growing out from expiants.

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Abbreviations

CA:

catecholamines

NA:

noradrenaline

A:

adrenaline

DA:

dopamine

6-OHDA:

6-hydroxydopamine

5-OHDA:

5-hydroxydopamine

NGF:

nerve growth factor

PNMT:

phenylethanolamine N-methyltransferase

TH:

tyrosine hydroxylase

DBH:

dopamine β-hydroxylase

Ach:

acetylcholine

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Authors and Affiliations

  1. Department of Anatomy, University of Kiel, West Germany

    K. Unsicker

  2. Department of Anatomy, University College London, London, UK

    Julie H. Chamley

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  1. K. Unsicker
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Part of this study was carried out in the Department of Zoology, University of Melbourne, Australia. We wish to thank Professor G. Burnstock for providing us with laboratory facilities

Supported by grants from Deutsche Forschungsgemeinschaft, a Research Fellowship of the University of Melbourne (K.U.), and a grant from the Life Insurance Medical Research Fund of Australia and New Zealand (J.H.C.). Thanks are due to Janet McConnell, Marlis Rall and Ingrid Rietiens for excellent technical assistance, Heidi Waluk for the photographic work and Brigitte Schierhorn for typing the manuscript

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Unsicker, K., Chamley, J.H. Growth characteristics of postnatal rat adrenal medulla in culture. Cell Tissue Res. 177, 247–268 (1977). https://doi.org/10.1007/BF00221086

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