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Am J Physiol Gastrointest Liver Physiol 293: G1272-G1280, 2007. First published October 18, 2007; doi:10.1152/ajpgi.00134.2007
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LIVER AND BILIARY TRACT

Focal adhesion disassembly is an essential early event in hepatic stellate cell chemotaxis

Andrew C. Melton, Russell K. Soon, Jr., J. Genevieve Park, Luis Martinez, Gregory W. deHart, and Hal F. Yee, Jr.

Department of Medicine and Liver Center, University of California San Francisco, San Francisco, California

Submitted 22 March 2007 ; accepted in final form 15 October 2007

Chemotaxis (i.e., directed migration) of hepatic stellate cells to areas of inflammation is a requisite event in the liver's response to injury. Previous studies of signaling pathways that regulate stellate cell migration suggest a key role for focal adhesions, but the exact function of these protein complexes in motility remains unclear. Focal adhesions attach a cell to its substrate and therefore must be regulated in a highly coordinated manner during migration. To test the hypothesis that focal adhesion turnover is an essential early event for chemotaxis in stellate cells, we employed a live-cell imaging technique in which chemotaxis was induced by locally stimulating the tips of rat stellate cell protrusions with platelet-derived growth factor-BB (PDGF). Focal adhesions were visualized with an antibody directed against vinculin, a structural component of the focal adhesion complex. PDGF triggered rapid disassembly of adhesions within 6.25 min, subsequent reassembly by 12.5 min, and continued adhesion assembly in concert with the spreading protrusion until the completion of chemotaxis. Blockade of adhesion disassembly by growing cells on fibronectin or treatment with nocodazole prevented a chemotactic response to PDGF. Augmentation of adhesion disassembly with ML-7 enhanced the chemotactic response to PDGF. These data suggest that focal adhesion disassembly is an essential early event in stellate cell chemotaxis in response to PDGF.

platelet-derived growth factor; migration; microtubules; myosin light chain kinase; fibronectin


Address for reprint requests and other correspondence: H. F. Yee, Jr., San Francisco General Hospital, Bldg. 40, Rm. 4102, 1001 Potrero Ave., San Francisco, CA 94110 (e-mail: hyee{at}medsfgh.ucsf.edu)






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