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Originally published as MBC in Press, 10.1091/mbc.E02-08-0537 on June 13, 2003

Vol. 14, Issue 8, 3065-3081, August 2003

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Activity and Distribution of Paxillin, Focal Adhesion Kinase, and Cadherin Indicate Cooperative Roles during Zebrafish Morphogenesis

Bryan D. Crawford * {dagger}, Clarissa A. Henry * {ddagger}, Todd A. Clason *, Amanda L. Becker *, and Merrill B. Hille * § ||

* Department of Biology and Developmental Biology Center, University of Washington, Seattle, Washington 98195-1800; § Institute of Neuroscience, University of Oregon, Eugene, Oregon 97403-1254

Submitted August 27, 2002; Revised April 13, 2003; Accepted April 15, 2003
Monitoring Editor: Mark Ginsberg

We investigated the focal adhesion proteins paxillin and Fak, and the cell-cell adhesion protein cadherin in developing zebrafish (Danio rerio) embryos. Cadherins are expressed in presomitic mesoderm where they delineate cells. The initiation of somite formation coincides with an increase in the phosphorylation of Fak, and the accumulation of Fak, phosphorylated Fak, paxillin, and fibronectin at nascent somite boundaries. In the notochord, cadherins are expressed on cells during intercalation, and phosphorylated Fak accumulates in circumferential rings where the notochord cells contact laminin in the perichordal sheath. Subsequently, changes in the orientations of collagen fibers in the sheath suggest that Fak-mediated adhesion allows longitudinal expansion of the notochord, but not lateral expansion, resulting in notochord elongation. Novel observations showed that focal adhesion kinase and paxillin concentrate at sites of cell-cell adhesion in the epithelial enveloping layer and may associate with actin cytoskeleton at epithelial junctions containing cadherins. Fak is phosphorylated at these epithelial junctions but is not phosphorylated on Tyr397, implicating a noncanonical mechanism of regulation. These data suggest that Fak and paxillin may function in the integration of cadherin-based and integrin-based cell adhesion during the morphogenesis of the early zebrafish embryo.

Abbreviations used: CAK-{beta}, cell adhesion kinase-{beta}, which is also known as protein tyrosine kinase 2{beta} (PTK2{beta}), and focal adhesion kinase 2 (FADK 2); ECM, extracellular matrix; EVL, enveloping layer; FAK, mouse, Xenopus, or human focal adhesion kinase protein; Fak, zebrafish focal adhesion kinase protein; all numbering of Fak residues is according to the known phosphorylated human sites; fak, mRNA or DNA focal adhesions kinase gene; hpf, hours postfertilization; PGMT, postgastrula mesoderm transition; TEM, transmission electron microscopy.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E02-08-0537. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E02-08-0537.

{dagger} Present address: Department of Biological Sciences, CW 405, University of Alberta, Edmonton, Canada T6G 2E9.

{ddagger} Present address: Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720.

|| Corresponding author. E-mail address: mbhille{at}u.washington.edu.




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