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Journal of Neuroscience, Vol 13, 5126-5138, Copyright © 1993 by Society for Neuroscience
A comparison of two immediate-early genes, c-fos and NGFI-B, as markers for functional activation in stress-related neuroendocrine circuitry
RK Chan, ER Brown, A Ericsson, KJ Kovacs and PE Sawchenko
Laboratory of Neuronal Structure and Function, Salk Institute, San Diego, California 92186-5800.
The promoter regions of the rat corticotropin-releasing factor (CRF),
oxytocin (OT), and vasopressin (AVP) genes contain sequences similar to the
cis-acting response element identified for NGFI-B, an immediate- early gene
structurally related to the steroid hormone receptor superfamily. Combined
immuno- and hybridization histochemical approaches were used to determine
whether challenges that influence the synthesis and secretion of CRF, OT,
and/or AVP result in altered expression in neurosecretory neurons of NGFI-B
and another immediate- early gene, c-fos, which is widely used as a marker
for functionally activated neurons. NGFI-B mRNA was found to be expressed
at constitutively high levels in the telencephalon, but not in the
endocrine hypothalamus, of unperturbed controls; basal levels of c-fos
expression were uniformly low throughout the CNS. NGFI-B and c-fos mRNAs,
and Fos protein, were induced with a similar time course and in similar
neuroendocrine cell types in response to acute hypotensive hemorrhage (15%
reduction in blood volume), intravenous injection of interleukin-1 beta
(IL-1 beta; 1.87 micrograms/kg), chronic salt loading (7 d maintenance on
2% saline), and acute bilateral adrenalectomy. c-fos mRNA and Fos protein
were readily demonstrable in afferent pathways that have been implicated as
mediating the neuroendocrine responses in the three stress paradigms; these
include medullary catecholaminergic cell groups in response to IL-1 beta
and hemorrhage, and cell groups lining the lamina terminalis in response to
salt loading. Challenge-specific induction of NGFI-B expression was
detectable in these extrahypothalamic cell groups, though with a lesser
sensitivity than that required to reveal NGFI-B induction in the
hypothalamus, or c-fos expression in these related afferents. These results
establish NGFI-B as a useful adjunct to c-fos, for revealing synaptic
and/or transcriptional activation in the magno- and parvocellular
neurosecretory systems. Differences in the sensitivity of the two markers
in revealing functionally related activation in extrahypothalamic regions
speak to general issues concerning the use of immediate-early genes in
mapping functional circuitry in the CNS.
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