Surfactant protein-A plays an important role in lung surfactant clearance: evidence using the surfactant protein-A gene-targeted mouse.
Previous studies with the isolated perfused rat lung showed that both clathrin- and actin-mediated pathways are responsible for endocytosis of dipalmitoylphosphatidylcholine (DPPC)-labeled liposomes by granular pneumocytes in the intact lung. Using surfactant protein-A (SP-A) gene-targeted mice, we examined the uptake of [(3)H]DPPC liposomes by isolated mouse lungs under basal and secretagogue-stimulated conditions. Unilamellar liposomes composed of [(3)H]DPPC: phosphatidylcholine:cholesterol:egg phosphatidylglycerol (10:5:3:2 mol fraction) were instilled into the trachea of anesthetized mice, and the lungs were perfused (2 h). Uptake was calculated as percentage of instilled disintegrations per minute in the postlavaged lung. Amantadine, an inhibitor of clathrin and, thus, receptor-mediated endocytosis via clathrin-coated pits, decreased basal [(3)H]DPPC uptake by 70% in SP-A +/+ but only by 20% in SP-A -/- lung, data compatible with an SP-A/receptor-regulated lipid clearance pathway in the SP-A +/+ mice. The nonclathrin, actin-dependent process was low in the SP-A +/+ lung but accounted for 55% of liposome endocytosis in the SP-A -/- mouse. With secretagogue (8-bromoadenosine 3',5'-cyclic monophosphate) treatment, both clathrin- and actin-dependent lipid clearance were elevated in the SP-A +/+ lungs while neither pathway responded in the SP-A -/- lungs. Binding of iodinated SP-A to type II cells isolated from both genotypes of mice was similar indicating a normal SP-A receptor status in the SP-A -/- lung. Inclusion of SP-A with instilled liposomes served to "rescue" the SP-A -/- lungs by reestablishing secretagogue-dependent enhancement of liposome uptake. These data are compatible with a major role for receptor-mediated endocytosis of DPPC by granular pneumocytes, a process critically dependent on SP-A.
Bates
Sandra R
SR
Institute for Environmental Medicine, University of Pennsylvania School of Medicine, 3620 Hamilton Walk, Philadelphia, PA 19104-6068, USA. batekenn@mail.med.upenn.edu
Dodia
Chandra
C
Tao
Jian-Qin
JQ
Fisher
Aron B
AB
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D002794:Choline / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism; D015638:Cytochalasin D / Q000494:pharmacology; D018390:Gene Targeting; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008081:Liposomes; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010477:Perfusion; D011485:Protein Binding / Q000187:drug effects; D011650:Pulmonary Alveoli / Q000187:drug effects / Q000378:metabolism; D037662:Pulmonary Surfactant-Associated Protein A / Q000172:deficiency* / Q000378:metabolism*; D011663:Pulmonary Surfactants / Q000378:metabolism*; D014316:Tritium
D000199:Actins; D002966:Clathrin; D007457:Iodine Radioisotopes; D008081:Liposomes; D037662:Pulmonary Surfactant-Associated Protein A; D011663:Pulmonary Surfactants; D014316:Tritium; D015638:Cytochalasin D; D015060:1,2-Dipalmitoylphosphatidylcholine; D002794:Choline