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JBC, Vol. 252, Issue 11, 3870-3875, Jun, 1977
L. Y. Yu, R. J. Tushinski and F. C. Bancroft
Conditions were determined for measuring growth hormone synthesis by a
clonal strain of rat pituitary cells grown in suspension culture.
Incubation of the cells with [3H]leucine in either continuous labeling or
pulse-chase experiments showed that secretion of newly synthesized growth
hormone commences only after a lag of about 15 min. The pulse-chase
experiments also demonstrated that there is no detectable degradation by
the cells of growth hormone. Thus growth hormone synthesis could be
measured, in the absence of complications arising either from secretion or
degradation of growth hormone, by incubating the cells with [3H]leucine for
10 min. Exposure of cells grown under the usual culture conditions to
dexamethasone (a synthetic glucocorticoid) led to an average stimulation of
specific growth hormone synthesis (growth hormone synthesis/total
cytoplasmic protein synthesis) of only 2.6-fold. However, two other growth
conditions were found in which dexamethasone routinely yielded a 5- to
15-fold stimulation of specific growth hormone synthesis. One of these
conditions, involving substitution of 10% fetal calf serum for the normal
serum supplement, was employed in subsequent experiments. A stimulation of
specific growth hormone synthesis could be observed at 10(-9) M
dexamethasone, and the maximum stimulation was observed at dexamethasone
concentrations of about 10(-8) to 10(-7) M. There was a lag of about 6 h
before a stimulation by dexamethasone of specific growth hormone synthesis
was detected. Thereafter, the stimulation increased in a nearly linear
fashion until maximum stimulation was reached at about 48 h.
Glucocorticoid induction of growth hormone synthesis in a strain of rat pituitary cells
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