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Journal of Neuroscience, Vol 11, 2321-2331, Copyright © 1991 by Society for Neuroscience
c-fos mRNA, Fos, and Fos-related antigen induction by hypertonic saline and stress
FR Sharp, SM Sagar, K Hicks, D Lowenstein and K Hisanaga
Department of Neurology, University of California, San Francisco.
The induction of c-fos mRNA was assessed using Northern blots and in situ
hybridization in adult rats administered hypertonic saline (HS) and
isotonic saline (IS). HS induced c-fos mRNA in magnocellular
paraventricular nucleus (PVNm), parvocellular paraventricular nucleus
(PVNp), supraoptic nucleus (SON), and lamina terminalis (LMT). This
occurred within 5 min, peaked at 30-60 min, and disappeared by 180 min. Fos
protein, detected using a specific monoclonal antibody, was maximal at 1-2
hr and disappeared 4-8 hr after HS administration. This confirms
observations showing that the c-fos gene response is transient even in the
presence of a continuing stimulus. In contrast, Fos-like immunoreactivity
(FLI), detected using two polyclonal antisera, was observed in PVNm, PVNp,
SON, and LMT for 1-24 hr during continuous osmotic stimulation. Moreover,
FLI was observable in these structures for 7 d in rats administered HS and
allowed to drink water ad libitum beginning 24 hr later. At times greater
than 8 hr, FLI presumably represents Fos-related antigens (FRA), proteins
immunologically and functionally related to Fos, whose expression is much
more prolonged than authentic Fos following the osmotic stimulus. In
addition to induction of c-fos expression in regions specifically involved
in osmotic regulation, HS injections also induced c-fos in many other
forebrain regions. In order to assess the induction of c-fos mRNA due to
the "stress" of the injections, rats injected with isotonic saline were
compared to uninjected controls. Isotonic saline injections induced c-fos
mRNA in the PVNp, anterior hypothalamus, suprachiasmatic nucleus, cingulate
gyrus, neocortex, ventral lateral septal nucleus, piriform cortex,
hippocampal pyramidal and dentate granule neurons, paraventricular and
intralaminar thalamic nuclei, bed nuclei of stria terminalis, cortical and
medial amygdaloid nuclei, and other structures. In accord with other work,
we interpret this pattern of c- fos expression to result from the stress of
handling and injections. Since Fos and FRA probably bind to the promoters
of target genes and regulate their expression, they likely mediate
biochemical changes in the cells activated by the osmotic and stressful
stimuli. Whereas the Fos signal is transient, FRA may act on target genes
for the duration of the stimulus or longer.
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