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Published online 23 February 2004. doi:10.1083/jcb.200308104
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 5, 717-727
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Article

Mammalian PAR-1 determines epithelial lumen polarity by organizing the microtubule cytoskeleton



David Cohen1, Patrick J. Brennwald2, Enrique Rodriguez-Boulan1, and Anne Müsch1

1 Margaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, New York, NY 10021
2 Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, NC 27599

Address correspondence to Anne Müsch, Margaret M. Dyson Vision Research Institute, Weill Medical College of Cornell University, 1300 York Ave., Box 233, New York, NY 10021. Tel.: (212) 746-2260. Fax: (212) 746-8101. email: amuesch{at}mail.med.conell.edu

Epithelial differentiation involves the generation of luminal surfaces and of a noncentrosomal microtubule (MT) network aligned along the polarity axis. Columnar epithelia (e.g., kidney, intestine, and Madin-Darby canine kidney [MDCK] cells) generate apical lumina and orient MT vertically, whereas liver epithelial cells (hepatocytes and WIFB9 cells) generate lumina at cell–cell contact sites (bile canaliculi) and orient MTs horizontally. We report that knockdown or inhibition of the mammalian orthologue of Caenorhabditis elegans Par-1 (EMK1 and MARK2) during polarization of cultured MDCK and WIFB9 cells prevented development of their characteristic lumen and nonradial MT networks. Conversely, EMK1 overexpression induced the appearance of intercellular lumina and horizontal MT arrays in MDCK cells, making EMK1 the first known candidate to regulate the developmental branching decision between hepatic and columnar epithelial cells. Our experiments suggest that EMK1 primarily promotes reorganization of the MT network, consistent with the MT-regulating role of this gene product in other systems, which in turn controls lumen formation and position.

Key Words: EMK1; MARK2; MDCK; WIFB; apical surface

The online version of this article includes supplemental material.

Abbreviations: BC, bile canaliculi; DPPIV, dipeptidyl aminopeptidase IV; KN-EMK1, kinase-negative EMK1; MT, microtubule; MTOC, MT organizing center; VAC, vacuolar apical compartment.


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