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1 School of Biological
Sciences,
The aim of this study was to analyze the effects
of osmotic shock and secretagogues such as ATP and
12-O-tetradecanoylphorbol 13-acetate
(TPA) on various intracellular signaling pathways in primary cultures
of alveolar type II cells and examine their potential role in
regulating events such as secretion and apoptosis in these cells.
Sorbitol-induced osmotic stress caused the sustained release of
[3H]phosphatidylcholine
([3H]PC) from primary cultures of rat alveolar type II
cells prelabeled with
[3H]choline chloride.
This release was not dependent on protein kinase C because
downregulation of the major protein kinase C isoforms (
,
II, 
, and 
)
expressed in alveolar type II cells had no effect on
[3H]PC secretion.
Sorbitol, as well as the known secretagogues TPA and ATP, activated
extracellular signal-regulated kinase. Although an inhibitor of the
extracellular signal-regulated kinase cascade, PD-98059, blocked this
activation, it had no effect on the release of
[3H]PC. Sorbitol and
ultraviolet C radiation, but not TPA or ATP, were also found to
activate both p38 and stress-activated protein kinase/c-Jun
NH2-terminal kinase. Furthermore,
both sorbitol and ultraviolet C radiation induced apoptosis in alveolar
type II cells as demonstrated by Hoechst 33258 staining of the
condensed nuclei, the generation of DNA ladders, and the activation of
caspases. The data indicate that multiple signaling pathways are
activated by traditional secretagogues such as TPA and ATP and by
cellular stresses such as osmotic shock and that these may be involved in regulating secretory and apoptotic events in alveolar type II cells.
sorbitol; 12-O-tetradecanoylphorbol 13-acetate; adenosine 5'-triphosphate; mitogen-activated protein kinase; stress-activated signaling
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