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Am J Physiol Lung Cell Mol Physiol 277: L893-L900, 1999;
1040-0605/99 $5.00
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Vol. 277, Issue 5, L893-L900, November 1999

Threshold calcium levels for lamellar body exocytosis in type II pneumocytes

Thomas Haller, Klaus Auktor, Manfred Frick, Norbert Mair, and Paul Dietl

Department of Physiology, University of Innsbruck, A-6020 Innsbruck, Austria

Pulmonary surfactant is secreted via exocytosis of lamellar bodies (LBs) by alveolar type II cells. Here we analyzed the dependence of LB exocytosis on intracellular Ca2+ concentration ([Ca2+]i). In fura 2-loaded cells, [Ca2+]i was selectively elevated by flash photolysis of a cell-permeant caged Ca2+ compound (o-nitrophenyl EGTA-AM) or by gradually enhancing cellular Ca2+ influx. Simultaneously, surfactant secretion by single cells was analyzed with the fluorescent dye FM 1-43, enabling detection of exocytotic events with a high temporal resolution (T. Haller, J. Ortmayr, F. Friedrich, H. Volkl, and P. Dietl. Proc. Natl. Acad. Sci. USA 95: 1579-1584, 1998). Exocytosis was initiated at a threshold concentration near 320 nmol/l with both instantaneous or gradual [Ca2+]i elevations. The exocytotic response to flash photolysis was highest during the first minute after the rise in [Ca2+]i and thus almost identical to purinoceptor stimulation by ATP. Correspondingly, the effects of ATP on initial secretion could be sufficiently explained by its ability to mobilize Ca2+. This was further demonstrated by the fact that exocytosis is significantly blocked by suppression of the ATP-induced Ca2+ signal below ~300 nmol/l. Our results suggest a highly Ca2+-sensitive step in LB exocytosis.

surfactant secretion; alveolar type II cells; flash photolysis; caged calcium


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