p114RhoGEF governs cell motility and lumen formation during tubulogenesis through a ROCK-myosin-II pathway.
Tubulogenesis is fundamental to the development of many epithelial organs. Although lumen formation in cysts has received considerable attention, less is known about lumenogenesis in tubes. Here, we utilized tubulogenesis induced by hepatocyte growth factor (HGF) in MDCK cells, which form tubes enclosing a single lumen. We report the mechanism that controls tubular lumenogenesis and limits each tube to a single lumen. Knockdown of p114RhoGEF (also known as ARHGEF18), a guanine nucleotide exchange factor for RhoA, did not perturb the early stages of tubulogenesis induced by HGF. However, this knockdown impaired later stages of tubulogenesis, resulting in multiple lumens in a tube. Inhibition of Rho kinase (ROCK) or myosin IIA, which are downstream of RhoA, led to formation of multiple lumens. We studied lumen formation by live-cell imaging, which revealed that inhibition of this pathway blocked cell movement, suggesting that cell movement is necessary for consolidating multiple lumens into a single lumen. Lumen formation in tubules is mechanistically quite different from lumenogenesis in cysts. Thus, we demonstrate a new pathway that regulates directed cell migration and formation of a single lumen during epithelial tube morphogenesis.
Department of Anatomy, University of California, San Francisco, CA 94158, USA.
Minji
M
Kim
School of Chemistry & Molecular Bioscience, University of Queensland, Brisbane, QLD 4072, Australia.
Annette
A
M Shewan
Department of Cell Biology, Johns Hopkins University, Baltimore, MD 21205, USA.
Andrew J
AJ
Ewald
Department of Anatomy, University of California, San Francisco, CA 94158, USA.
Zena
Z
Werb
Department of Anatomy, University of California, San Francisco, CA 94158, USA Department of Biochemistry/Biophysics, University of California, San Francisco, CA 94158, USA keith.mostov@ucsf.edu.
Keith E
KE
Mostov
D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D024462:Myosin Type II / Q000235:genetics / Q000378:metabolism*; D064067:Rho Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000235:genetics / Q000378:metabolism*
D064067:Rho Guanine Nucleotide Exchange Factors; D054460:rho-Associated Kinases; D024462:Myosin Type II
Epithelia; Hepatocyte growth factor; Madin–Darby canine kidney cells; Migration; Tube