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UNC-6 (netrin) orients the invasive membrane of the anchor cell in C. elegans

Nature Cell Biology volume 11, pages 183–189 (2009)Cite this article

Abstract

Despite their profound importance in the development of cancer, the extracellular cues that target cell invasion through basement membrane barriers remain poorly understood1. A central obstacle has been the difficulty of studying the interactions between invading cells and basement membranes in vivo2,3. Using the genetically and visually tractable model of Caenorhabditis elegans anchor cell (AC) invasion, we show that UNC-6 (netrin) signalling, a pathway not previously implicated in controlling cell invasion in vivo, is a key regulator of this process. Site of action studies reveal that before invasion, localized UNC-6 secretion directs its receptor, UNC-40, to the plasma membrane of the AC, in contact with the basement membrane. There, UNC-40 polarizes a specialized invasive membrane domain through the enrichment of actin regulators, F-actin and phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). Cell ablation experiments indicate that UNC-6 promotes the formation of invasive protrusions from the AC that break down the basement membrane in response to a subsequent vulval cue. Together, these results characterize an invasive membrane domain in vivo, and reveal a role for UNC-6 (netrin) in polarizing this domain towards its basement membrane target.

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Figure 1: The AC fails to invade in unc-6 mutants.
Figure 2: UNC-6 is a distinct VNC-derived, pro-invasive cue.
Figure 3: UNC-6 directs its receptor UNC-40 to the invasive cell membrane.
Figure 4: UNC-6 localizes the Rac protein MIG-2, F-actin and PI(4,5)P2 to the invasive cell membrane.
Figure 5: UNC-6 promotes the deposition of HIM-4 at the site of invasion.

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Acknowledgements

We are grateful to C. Bargmann for the GFP::unc-34 and unc-40::GFP vectors, the hs>unc-6::HA integrated strain, and advice; W. Wadsworth for the Lam-1::GFP vector; E. Lundquist for the GFP::ced-10 vector; Y. Goshima for Venus::unc-6 strains; D. Kiehart for the moeADB::GFP vector; the Caenorhabditis Genetics Center for providing strains; and N. Sherwood, Z. Wang, G. Lopez, G. Miley and D. Matus for comments on the manuscript. This work was supported by a Basil O'Connor Award, Pew Scholars Award, and NIH Grant GM079320 to D.R.S.

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Authors and Affiliations

  1. Department of Biology, Duke University, Box 90338, Durham, 27708, NC, USA

    Joshua W. Ziel, Elliott J. Hagedorn & David R. Sherwood

  2. Department of Biomolecular Chemistry, University of Wisconsin-Madison, 1300 University Avenue, Madison, 53706, WI, USA

    Anjon Audhya

  3. Molecular Cancer Biology Program, Duke University Medical Center, Durham, 27708, NC, USA

    David R. Sherwood

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Contributions

A.A., D.R.S. and J.W. Z. generated the fluorescent markers, C. elegans transgenic lines and genetic stocks used in this study. The AC invasion phenotypes in unc-6 and unc-40 mutants were originally observed by D.R.S.; D.R.S. and J.W.Z. designed the experiments described in the paper; D.R.S., E.J.H. and J.W.Z. performed the experiments and analysed the data.

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Correspondence to David R. Sherwood.

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The authors declare no competing financial interests.

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Ziel, J., Hagedorn, E., Audhya, A. et al. UNC-6 (netrin) orients the invasive membrane of the anchor cell in C. elegans. Nat Cell Biol 11, 183–189 (2009). https://doi.org/10.1038/ncb1825

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