Immunocytochemical localization and identification of the major surfactant protein in adult rat lung.
We investigated the cellular and subcellular sites of metabolism of the 72,000 dalton protein of pulmonary surfactant in order to provide insights into mechanisms of synthesis, intracellular assembly, and intraalveolar metabolism of this phospholipid-rich secretory product. Surfactant (approximately 90% lipid, 10% protein by weight) was purified by density gradient centrifugation of material obtained by lavaging rat lungs. The purified material was used to generate an antiserum from which a specific antibody was obtained by affinity chromatography. A horseradish peroxidase-labeled Fab was used to localize the antigen in rat lung. The antibody labeled the rough endoplasmic reticulum and Golgi apparatus of type II cells only. Some multivesicular bodies in type II cells were also labeled, but whether the antigen was present in lamellar bodies was uncertain. Phagosomes of alveolar macrophages were labeled as were similar inclusions in type I cells. Using indirect immunocytochemistry we determined that the labeling of alveolar cell surfaces does not represent the presence of a continuous layer of secreted surfactant. These results suggest that only the type II cell synthesizes surfactant protein and than mainly alveolar macrophages participate in its catabolism. The initial intracellular site of the association of protein with lipid may be multivesicular bodies as suggested previously by others.
D000818:Animals; D005260:Female; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments; D008168:Lung / Q000032:analysis* / Q000166:cytology / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D011510:Proteolipids / Q000032:analysis*; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants / Q000032:analysis*; D051381:Rats; D013347:Subcellular Fractions / Q000648:ultrastructure
D007140:Immunoglobulin Fab Fragments; D011510:Proteolipids; D037661:Pulmonary Surfactant-Associated Proteins; D011663:Pulmonary Surfactants; D006735:Horseradish Peroxidase