title: F-actin scaffold stabilizes lamellar bodies during surfactant secretion.
abstract: Alveolar type 2 (AT2) cells secrete surfactant that forms a protective layer on the lung's alveolar epithelium. Vesicles called lamellar bodies (LBs) store surfactant. Failure of surfactant secretion, which causes severe lung disease, relates to the manner in which LBs undergo exocytosis during the secretion. However, the dynamics of LBs during the secretion process are not known in intact alveoli. Here, we addressed this question through real-time confocal microscopy of single AT2 cells in live alveoli of mouse lungs. Using a combination of phospholipid and aqueous fluorophores that localize to LBs, we induced surfactant secretion by transiently hyperinflating the lung, and we quantified the secretion in terms of loss of bulk LB fluorescence. In addition, we quantified inter-LB phospholipid flow through determinations of fluorescence recovery after photobleaching. Furthermore, we determined the role of F-actin in surfactant secretion through expression of the fluorescent F-actin probe Lifeact. Our findings indicate that, in AT2 cells in situ, LBs are held in an F-actin scaffold. Although F-actin transiently decreases during surfactant secretion, the LBs remain stationary, forming a chain of vesicles connected by intervesicular channels that convey surfactant to the secretion site on the plasma membrane. This is the first instance of a secretory process in which the secretory vesicles are immobile, but form a conduit for the secretory material.
authors.affiliation: 630 West 168th St., BB 8-812, New York, NY 10032. jb39@columbia.edu.
authors.forename: Mohammad N
authors.initials: MN
authors.lastname: Islam
authors.forename: Galina A
authors.initials: GA
authors.lastname: Gusarova
authors.forename: Eiji
authors.initials: E
authors.lastname: Monma
authors.forename: Shonit R
authors.initials: SR
authors.lastname: Das
authors.forename: Jahar
authors.initials: J
authors.lastname: Bhattacharya
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D056809:Alveolar Epithelial Cells / Q000378:metabolism*; D000818:Animals; D051379:Mice; D055503:Protein Multimerization; D037701:Pulmonary Surfactant-Associated Protein B / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D055571:Secretory Pathway; D022142:Secretory Vesicles / Q000378:metabolism*; D046509:Tissue Culture Techniques
chemical_list: D000199:Actins; D037701:Pulmonary Surfactant-Associated Protein B
keywords: alveolar type 2 cells; fluorescence microscopy; lung injury