title: Laminin-6 assembles into multimolecular fibrillar complexes with perlecan and participates in mechanical-signal transduction via a dystroglycan-dependent, integrin-independent mechanism.
abstract: Mechanical ventilation is a valuable treatment regimen for respiratory failure. However, mechanical ventilation (especially with high tidal volumes) is implicated in the initiation and/or exacerbation of lung injury. Hence, it is important to understand how the cells that line the inner surface of the lung [alveolar epithelial cells (AECs)] sense cyclic stretching. Here, we tested the hypothesis that matrix molecules, via their interaction with surface receptors, transduce mechanical signals in AECs. We first determined that rat AECs secrete an extracellular matrix (ECM) rich in anastamosing fibers composed of the alpha3 laminin subunit, complexed with beta1 and gamma1 laminin subunits (i.e. laminin-6), and perlecan by a combination of immunofluorescence microscopy and immunoblotting analyses. The fibrous network exhibits isotropic expansion when exposed to cyclic stretching (30 cycles per minute, 10% strain). Moreover, this same stretching regimen activates mitogen-activated-protein kinase (MAPK) in AECs. Stretch-induced MAPK activation is not inhibited in AECs treated with antagonists to alpha3 or beta1 integrin. However, MAPK activation is significantly reduced in cells treated with function-inhibiting antibodies against the alpha3 laminin subunit and dystroglycan, and when dystroglycan is knocked down in AECs using short hairpin RNA. In summary, our results support a novel mechanism by which laminin-6, via interaction with dystroglycan, transduces a mechanical signal initiated by stretching that subsequently activates the MAPK pathway in rat AECs. These results are the first to indicate a function for laminin-6. They also provide novel insight into the role of the pericellular environment in dictating the response of epithelial cells to mechanical stimulation and have broad implications for the pathophysiology of lung injury.
authors.affiliation: Division of Pulmonary Medicine, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA. j-jones3@northwestern.edu
authors.forename: Jonathan C R
authors.initials: JC
authors.lastname: Jones
authors.forename: Kimberly
authors.initials: K
authors.lastname: Lane
authors.forename: Susan B
authors.initials: SB
authors.lastname: Hopkinson
authors.forename: Emilia
authors.initials: E
authors.lastname: Lecuona
authors.forename: Robert C
authors.initials: RC
authors.lastname: Geiger
authors.forename: David A
authors.initials: DA
authors.lastname: Dean
authors.forename: Eduardo
authors.initials: E
authors.lastname: Correa-Meyer
authors.forename: Meredith
authors.initials: M
authors.lastname: Gonzales
authors.forename: Kevin
authors.initials: K
authors.lastname: Campbell
authors.forename: Jacob I
authors.initials: JI
authors.lastname: Sznajder
authors.forename: Scott
authors.initials: S
authors.lastname: Budinger
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D049030:Dystroglycans / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism*; D007797:Laminin / Q000737:chemistry* / Q000378:metabolism*; D008168:Lung / Q000166:cytology; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction
chemical_list: D019812:Heparan Sulfate Proteoglycans; D016023:Integrins; D007797:Laminin; D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; D021122:Protein Subunits; C119337:laminin 6; C029967:nidogen; C071927:perlecan; D049030:Dystroglycans