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Journal of Neuroscience, Vol 5, 2119-2142, Copyright © 1985 by Society for Neuroscience
Environmental influences in the development of neural crest derivatives: glucocorticoids, growth factors, and chromaffin cell plasticity
AJ Doupe, SC Landis and PH Patterson
The neural crest gives rise to three major adrenergic cell types:
sympathetic principal neurons, adrenal chromaffin cells, and small
intensely fluorescent (SIF) cells. All of these derivatives synthesize and
store catecholamines, but they differ in numerous other characteristics.
SIF cells appear intermediate in phenotype between the other two. We have
examined the role of several environmental factors in the differentiation
of sympathetic principal neurons and adrenal chromaffin cells. In previous
studies of young rat adrenal chromaffin cells in dissociated cell culture,
differentiated characteristics such as the presence of the enzyme
phenylethanolamine N-methyltransferase (PNMT), epinephrine (E) synthesis,
and large catecholamine storage vesicles were not well maintained. Here we
describe long-term culture of chromaffin cells which, in the presence of
micromolar glucocorticoid, maintained all of these characteristics. In
addition, chromaffin cells of a variety of ages were found to be dependent
on glucocorticoid for long-term survival in culture. In the absence of
glucocorticoid, many adrenal chromaffin cells from neonatal rats could be
rescued by nerve growth factor (NGF) administration. They extended
neurites, as previously described by Unsicker and colleagues (Unsicker, K.,
B. Krisch, U. Otten, and H. Thoenen (1978) Proc. Natl. Acad. Sci. U.S.A.
75: 3498-3502). In contrast to previous studies, however, with long-term
exposure to NGF these cells became indistinguishable from mature
sympathetic neurons, as judged by the following morphological and
biochemical criteria: increased cell size and loss of intense CA
fluorescence in their cell bodies; acquisition of characteristic neuronal
ultrastructure, including morphologically specialized synapses; loss of
chromaffin granules, PNMT, and E synthesis; and acquisition of neuron
markers, including tetanus toxin labeling and immunoreactivity to
neurofilament protein. This conversion to neurons was markedly enhanced by
addition of a non-neuronal cell conditioned medium (CM) containing a
neurite-promoting factor, which acted by increasing the NGF responsiveness
of the chromaffin cells. Even chromaffin cells from adult rats, which are
known to grow few processes in response to NGF alone, became neuronal in
the presence of this CM plus NGF. While converting to neurons, adrenal
chromaffin cells transiently assumed an intermediate phenotype resembling
type I SIF cells, which suggests particular developmental relationships
between the different cell types of the sympathoadrenal lineage.(ABSTRACT
TRUNCATED AT 400 WORDS)
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