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Am J Physiol Cell Physiol (September 1, 2004). doi:10.1152/ajpcell.00397.2004
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Submitted on August 13, 2004
Accepted on August 30, 2004

Barrier Role of Actin Filaments in Regulated Mucin Secretion from Airway Goblet Cells

Camille Ehre1, Andrea H. Rossi1, Lubna H. Abdullah1, Kathleen De Pestel1, Sandra Hill1, John C. Olsen1, and C. William Davis1*

1 Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina, Chapel Hill, NC, USA

* To whom correspondence should be addressed. E-mail: cwdavis{at}med.unc.edu.

Airway goblet cells secrete mucin onto mucosal surfaces under regulation of an apical, PLC/Gq-coupled P2Y2 receptor. We tested whether cortical actin filaments negatively regulate exocytosis in goblet cells by forming a barrier between secretory granules and plasma membrane docking sites, as postulated for other secretory cells. Immunostaining of human lung tissues and SPOC1 cells (an epithelial, mucin-secreting cell line) revealed an apical distribution of {beta}- and {gamma}-actin in ciliated and goblet cells. In goblet cells, actin appeared as a prominent subplasmalemmal sheet lying between granules and the apical membrane, and it disappeared from SPOC1 cells activated by purinergic agonist. Disruption of actin filaments with latrunculin A stimulated SPOC1 cell mucin secretion under basal and agonist-activated conditions, whereas stabilization with jasplakinolide, or overexpression of YFP-{beta}- or YFP-{gamma}-actin inhibited secretion. MARCKS, a PKC-activated actin-plasma membrane tethering protein, was phosphorylated following agonist stimulation, suggesting a translocation to the cytosol. Scinderin (= adseverin), a Ca2+-activated actin filament severing and capping protein was cloned from human airway and SPOC1 cells, and synthetic peptides corresponding to its actin-binding domains inhibited mucin secretion. We conclude that actin filaments negatively regulate mucin secretion, basally, in airway goblet cells, and are dynamically remodeled in agonist-stimulated cells to promote exocytosis.




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