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An integrin–ILK–microtubule network orients cell polarity and lumen formation in glandular epithelium

Nature Cell Biology volume 15, pages 17–27 (2013)Cite this article

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Abstract

The extracellular matrix has a crucial role in determining the spatial orientation of epithelial polarity and the formation of lumens in glandular tissues; however, the underlying mechanisms remain elusive. By using Cre–Lox deletion we show that β1 integrins are required for normal mammary gland morphogenesis and lumen formation, both in vivo and in a three-dimensional primary culture model in which epithelial cells directly contact a basement membrane. Downstream of basement membrane β1 integrins, Rac1 is not involved; however, ILK is needed to polarize microtubule plus ends at the basolateral membrane and disrupting each of these components prevents lumen formation. The integrin–microtubule axis is necessary for the endocytic removal of apical proteins from the basement-membrane–cell interface and for internal Golgi positioning. We propose that this integrin signalling network controls the delivery of apical components to the correct surface and thereby governs the orientation of polarity and development of lumens.

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Figure 1: Deletion of β1 integrins or ILK disrupts acinar morphogenesis.
Figure 2: Luminal filling is not due to a lack of apoptosis.
Figure 3: Apical polarity is inverted in β1 integrin and ILK-KO acini.
Figure 4: β1 integrins and ILK control internal Golgi polarity.
Figure 5: β1 integrins and ILK control polarity and lumens through polarization of microtubules.
Figure 6: Dynamic microtubules are required for apical relocation of aPKC and lumen formation.
Figure 7: β1 integrins orient polarity through an endocytic mechanism.
Figure 8: β1 integrin signalling specifies the orientation and maintenance of epithelial polarity and the formation of lumens.

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Acknowledgements

We thank C. Jones for doing the electron microscopy, S. Schmid (Scripps Research Institute, La Jolla, USA) and B. Ceresa (University of Louisville, USA) for dynamin and Rab5a adenoviruses, C. Wu (University of Pittsburgh, USA) for anti-ILK monoclonal, M. Lowe (University of Manchester, UK) for GM130 polyclonal and P. March for confocal microscope training. The FLS Bioimaging Facility microscopes were purchased with grants from the BBSRC, Wellcome and the University of Manchester Strategic Fund. Thanks to P. Caswell, T. Hardingham, M. Humphries, M. Lowe, N. Papalopulu and P. Lu and for critical appraisal of the manuscript. This work was supported by the Wellcome Trust (no. 081203/Z/06/Z). The Wellcome Trust Centre for Cell-Matrix Research is supported by core funding from the Wellcome Trust (no. 088785/Z/09/Z).

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  1. Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, and Manchester Breast Centre, University of Manchester, Oxford Road, Manchester M13 9PT, UK

    Nasreen Akhtar & Charles H. Streuli

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  1. Nasreen Akhtar
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N.A. conceived ideas, performed experiments, analysed and interpreted the data and wrote the manuscript. C.H.S. conceived ideas and wrote the manuscript.

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Correspondence to Nasreen Akhtar or Charles H. Streuli.

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Wildtype MEC acinar on BMM rendered in 3D.

A third was removed to show an apical lumen (aPKC; red) and basolateral surface (β1 integrin; green). (MOV 11467 kb)

β1 integrin deleted MEC acinar on BMM rendered in 3D

A third was removed to show filled lumen. Note apical polarity (aPKC; red) inversion to outer membrane and a loss of β1 integrin (green). (MOV 9060 kb)

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Akhtar, N., Streuli, C. An integrin–ILK–microtubule network orients cell polarity and lumen formation in glandular epithelium. Nat Cell Biol 15, 17–27 (2013). https://doi.org/10.1038/ncb2646

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