Actin depolymerization is developmentally regulated in rat type II cells exposed to terbutaline.
The type II alveolar epithelial cell synthesizes and secretes pulmonary surfactant. Terbutaline enhances phospholipid release from adult and fetal type II cells. Our hypothesis is that the actin network of microfilaments regulates the secretory activity of the type II cell. To examine the developmental regulation of the changes in actin subfractions associated with secretory activity, cultures of type II cells derived from adult and 19-d fetal rat lung were incubated with or without 10 microM terbutaline for 1, 30, and 60 min. Dose-response effects of terbutaline were examined in adult type II cells. Effects of phorbol ester were also examined Globular (G-actin) and filamentous (F-actin) fractions were extracted from the cells and analyzed separately. Specified cellular equivalent volumes of each subfraction were analyzed by Western blotting, visualized by a color reaction, and quantified by densitometry. There was a decrease in the cytoskeletal F-actin pool along with an increase in the G-actin fraction within I min in adult type II cells exposed to terbutaline, indicating that depolymerization of F-actin occurs. Values returned to control levels by 60 min. In contrast, the decrease in F-actin, with a concomitant increase in G-actin, was maximal at 60 min in fetal cells exposed to terbutaline. There was a dose-dependent increase in actin depolymerization with maximal effects at 10 microM terbutaline. Phorbol ester also caused an increase in actin depolymerization. Depolymerization of the actin microfilament network may regulate transport and exocytosis of lamellar bodies in type II cells. We speculate that there is an early secretory mechanism that involves depolymerization of actin microfilaments and a late, actin-independent secretory mechanism present in adult type II cells. The timing of the response of the actin-dependent pathway is developmentally regulated. This may explain the developmental differences in the secretion of surfactant that we have previously shown.
Bhandari
V
V
Department of Pediatrics, University of Connecticut School of Medicine, Farmington 06030-2203, USA.
Lu
H
H
Pachter
J
J
Kresch
M J
MJ
D000199:Actins / Q000737:chemistry*; D000818:Animals; D001704:Biopolymers; D004353:Drug Evaluation, Preclinical; D008168:Lung / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D011663:Pulmonary Surfactants / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012636:Secretory Rate / Q000187:drug effects; D013726:Terbutaline / Q000627:therapeutic use*
D000199:Actins; D001704:Biopolymers; D011663:Pulmonary Surfactants; D013726:Terbutaline