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Volume 271, Number 41,
Issue of October 11, 1996
pp. 25277-25283
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
Secretagogues Increase the Expression of Surfactant Protein A
Receptors on Lung Type II Cells
(Received for publication, January 26, 1996, and in revised form, May 15, 1996)
Qiping
Chen
,
Sandra R.
Bates
and
Aron B.
Fisher
From the Institute for Environmental Medicine, University of
Pennsylvania School of Medicine,
Philadelphia, Pennsylvania 19104-6068
Since secretagogues have been shown to increase
the internalization of surfactant phospholipid and protein by lung
cells, we postulated that their action occurred through a mechanism
involving increased surfactant protein A (SP-A) receptor density.
Therefore, we evaluated the influence of secretagogues on the binding
of iodinated SP-A to alveolar type II cells. Type II cells were
isolated from rat lung and maintained in primary culture for 18 h
on Transwell membranes. Upon exposure to 8-bromo-cyclic AMP (cAMP, 0.1 mM), phorbol 12-myristate 13-acetate (PMA, 10 nM), terbutaline (0.1 mM), or ATP (1 mM), the binding of SP-A increased 1.5-2-fold. This
stimulation was cell substrate-dependent since type II
cells plated on plastic dishes did not show this effect. A time course
of the stimulation of SP-A binding due to secretagogues showed that
both cAMP and PMA increased SP-A binding by 2-fold after 20 min. With
cAMP, binding remained elevated for 2 h, while binding in the
presence of PMA had returned to control values. The effects of
submaximal concentrations of cAMP and PMA on binding were additive.
Inhibition of cellular protein synthesis with cycloheximide did not
alter the increase of SP-A binding stimulated by the secretagogues.
Type II cells pretreated with PMA responded to subsequent treatment
with cAMP by increasing SP-A binding, while these cells were refractory
to subsequent treatment with PMA. Both constitutive and regulated
binding of SP-A to type II cells were sensitive to trypsin. The binding
of SP-A to type II cells showed saturation at a concentration of 1 µg/ml SP-A under control and secretagogue-stimulated conditions, with
both total and calcium-dependent binding showing a 2-fold
increase upon secretagogue exposure. The data are consistent with the
hypothesis that secretagogues stimulate surfactant uptake, at least in
part, through recruitment of SP-A receptors to the type II cell
surface, resulting in an increase in the number of SP-A binding
sites.

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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
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