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MBC in Press, published online ahead of print June 14, 2006
Mol. Biol. Cell 10.1091/mbc.E05-11-1070

A more recent version of this article appeared on August 1, 2006
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Submitted on November 22, 2005
Revised on May 31, 2006
Accepted on June 1, 2006

Regulated Synthesis and Functions of Laminin 5 in Polarized Madin-Darby Canine Kidney Epithelial Cells

Grace Z. Mak,* Gina M. Kavanaugh,* Mary M. Buschmann,* Shaun M. Stickley,* Manuel Koch,{dagger} Kathleen Heppner Goss,* Holly Waechter,* Anna Zuk,{ddagger} and Karl S. Matlin*

*Laboratory of Epithelial Pathobiology, Department of Surgery, University of Cincinnati, Cincinnati, OH 45267; {dagger}Center for Biochemistry, Center for Molecular Medicine, and Department of Dermatology, University of Cologne, 50923 Cologne, Germany; {ddagger}Genzyme Corporation, Framingham, MA 01701

Monitoring Editor: Asma Nusrat

Renal tubular epithelial cells synthesize laminin 5 (LN5) during regeneration of the epithelium following ischemic injury. LN5 is a truncated laminin isoform of particular importance in the epidermis, but also constitutively expressed in a number of other epithelia. To investigate the role of LN5 in morphogenesis of a simple renal epithelium, we examined the synthesis and function of LN5 in the spreading, proliferation, wound-edge migration, and apical-basal polarization of Madin-Darby canine kidney (MDCK) cells. MDCK cells synthesize LN5 only when subconfluent, and degrade the existing LN5 matrix when confluent. Through the use of siRNA to knockdown the LN5 {alpha}3 subunit, we were able to demonstrate that LN5 is necessary for cell proliferation and efficient wound-edge migration, but not apical-basal polarization. Surprisingly, suppression of LN5 production caused cells to spread much more extensively than normal on uncoated surfaces, and exogenous keratinocyte LN5 was unable to rescue this phenotype. MDCK cells also synthesized laminin {alpha}5, a component of LN10, that independent studies suggest may form an assembled basal lamina important for polarization. Overall, our findings indicate that LN5 is likely to play an important role in regulating cell spreading, migration, and proliferation during reconstitution of a continuous epithelium.


Address correspondence to: Karl S. Matlin (karl.matlin{at}uc.edu)




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