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1, 
6 and 3 contribute to mechanical strain-induced differentiation of fetal type II cells via distinct mechanisms
1 Department of Pediatrics, Women & Infants Hospital, Providence, RI, USA
2 Department of Medicine, Rhode Island Hospital, Providence, RI, USA
3 Department of Pathology and Surgery, Children's Hospital, Boston, MA, USA
* To whom correspondence should be addressed. E-mail: jsanchezesteban{at}wihri.org.
Mechanical forces regulate lung maturation in the fetus by promoting type II epithelial differentiation. However, the cell surface receptors that transduce these mechanical cues into cellular responses remain largely unknown. When distal lung type II epithelial cells isolated from day E19 rat fetuses were cultured on flexible plates coated with laminin, fibronectin, vitronectin, collagen, or elastin and exposed to a level of mechanical strain (5%) similar to that observed in utero, transmembrane signaling responses were induced under all conditions, as measured by ERK activation. However, mechanical stress maximally increased expression of the type II cell differentiation marker, surfactant protein-C (SP-C), when cells were cultured on laminin substrates. Strain-induced alveolar epithelial differentiation was inhibited by interfering with cell binding to laminin using soluble laminin-peptides (IKVIV or YIGSR) or blocking antibodies against integrin 
1, 
3 or 6. Additional studies were carried out with substrates coated directly with different non-activating anti-integrin antibodies. Blocking integrin 1, and 6 binding sites inhibited both cell adhesion and differentiation, whereas inhibition of 3 prevented differentiation without altering cell attachment. These data demonstrate that various integrins contribute to mechanical control of type II lung epithelial cell differentiation on laminin substrates. However, they may act via distinct mechanisms, including some that are independent of their cell anchoring role.
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