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Am J Physiol Lung Cell Mol Physiol 278: L580-L590, 2000;
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Vol. 278, Issue 3, L580-L590, March 2000

Internalized SP-A and lipid are differentially resecreted by type II pneumocytes

Heide Wissel1, Stefan Zastrow1, Ekkehard Richter2, and Paul A. Stevens1

1 Clinic of Neonatology, University Children's Hospital Charité, Humboldt-University Berlin, 10098 Berlin; and 2 Institute of Biology, Department of Membrane Physiology, Humboldt-University Berlin, 10115 Berlin, Germany

Biochemical and morphological assays were developed to study surfactant protein A (SP-A) and lipid resecretion kinetics by isolated type II cells in vitro. After a 10-min uptake period with SP-A (3 µg/106 cells) in combination with liposomes (60 µg/106 cells), the cells were allowed to resecrete. After 5 min of resecretion, only 21.7 ± 4.6% of the internalized SP-A remained intracellularly compared with 54 ± 2.9% of the lipids. Extracellular SP-A present during the resecretion period partially inhibited resecretion (SP-A, 36% at 5 min; lipid, ~16% at 5 min). Lipid resecretion was also dependent on the SP-A concentration present during the uptake period. Although, as shown by confocal laser scanning microscopy, after a 10-min uptake period at 37°C, most of the fluorescein isothiocyanate-labeled SP-A and rhodamine-phosphatidylethanolamine-labeled lipids colocalized within the cells, after an additional 10 min of resecretion, both the strength of the fluorescence signals and the extent of colocalization had markedly decreased. These data indicate that internalized lipid and SP-A can be resecreted rapidly by type II cells, likely via different pathways.

surfactant protein A; liposomes; endocytosis; internalization


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