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J. Biol. Chem., Vol. 261, Issue 28, 12956-12959, 10, 1986
HH Zingg, D Lefebvre and G Almazan
Although the steps involved in biosynthesis and secretion of the
neuropeptide vasopressin (AVP) have been extensively studied, the factors
which regulate AVP gene expression remain unknown. Therefore, we sought to
determine the dynamics of AVP mRNA accumulation in response to a strong
stimulus for AVP release, i.e. during salt imbibition and the ensuing
period of rehydration. AVP mRNA levels were determined in terms of absolute
amounts by a novel quantitative densitometric hybridization assay, using in
vitro synthesized sense- strand RNA as a quantitative standard and
complementary anti-sense RNA as a specific probe. The template used for RNA
transcription consisted of a 196-base pair genomic DNA fragment
corresponding to exon C of the rat AVP gene. Determination of basal
hypothalamic AVP mRNA levels yielded 12.5 +/- 2.7 fmol/hypothalamus. Salt
imbibition, which induced a 6% rise in blood osmolality and an 82% loss of
pituitary AVP, resulted in a 3-fold increase of AVP mRNA to 35 +/- 5
fmol/hypothalamus. Following rehydration, plasma osmolality returned to
control levels by day 2, pituitary AVP by day 6, and hypothalamic AVP by
day 14. By contrast, AVP mRNA levels remained significantly elevated
throughout the 30-day rehydration period. Furthermore, pituitary AVP
reached a level of 177% of control by day 14 of rehydration. These data
show that osmotic stimulation results in a long-lasting elevation of
hypothalamic AVP mRNA; during rehydration, these elevated mRNA levels
direct AVP biosynthesis at a rate which surpasses secretory demands; AVP
mRNA accumulation does not appear to be directly regulated by either
pituitary or hypothalamic AVP. Therefore, either an unusually long
half-life of greater than or equal to 7 days must be assumed for AVP mRNA
or, alternatively, a continued stimulation of AVP gene transcription must
occur, even in the absence of a secretory stimulus and following complete
repletion of cellular AVP stores.
Regulation of vasopressin gene expression in rat hypothalamic neurons. Response to osmotic stimulation
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