Journal
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY
Volume 289, Issue 3, Pages L489-L496Publisher
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajplung.00074.2005
Keywords
pulmonary surfactant; alveolar epithelia; phosphatidylcholine; pneumocytes; adenosine 5'-triphosphate
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Funding
- NEI NIH HHS [EY-10009, R01 EY013434] Funding Source: Medline
- NHLBI NIH HHS [P01 HL019737-300018, P01-HL-019737, P01 HL019737, HL-57204] Funding Source: Medline
- NIGMS NIH HHS [R01 GM061012, R01 GM061012-06, GM-61012] Funding Source: Medline
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We developed a heterologous system to study the effect of mechanical deformation on alveolar epithelial cells. First, isolated primary rat alveolar type II (ATII) cells were plated onto silastic substrata coated with fibronectin and maintained in culture under conditions where they become alveolar type I-like (ATI) cells. This was followed by a second set of ATII cells labeled with the nontransferable, vital fluorescent stain 5-chloromethylfluorescein diacetate to distinguish them from ATI cells. By morphometric analysis, equibiaxial deformation ( stretch) of the silastic substratum induced comparable changes in cell surface area for both ATII and ATI cells. Surfactant lipid secretion was measured using cells metabolically labeled with [H-3] choline. In response to 21 % tonic stretch for 15 min, ATII cells seeded with ATI cells secreted nearly threefold more surfactant lipid compared with ATII cells seeded alone. ATI cells did not secrete lipid in response to stretch. The enhanced lipid secretion by ATII plus ATI cocultures was inhibited by treatment with apyrase and adenosine deaminase, suggesting that ATP release by ATI cells enhanced surfactant lipid secretion at 21 % stretch. This was confirmed using a luciferase assay where, in response to 21 % stretch, ATI cells released fourfold more ATP than ATII cells. Because ATI cells release significantly more ATP at a lower level of stretch than ATII cells, this supports the hypothesis that ATI cells are mechanosensors in the lung and that paracrine stimulation of ATII cells by extracellular ATP released from ATI cells plays a role in regulating surfactant secretion.
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