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TRPC1 channels regulate directionality of migrating cells

  • Signaling and Cell Physiology
  • Published:
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Abstract

Cell migration depends on the generation of structural asymmetry and on different steps: protrusion and adhesion at the front and traction and detachment at the rear part of the cell. The activity of Ca2+ channels coordinate these steps by arranging intracellular Ca2+ signals along the axis of movement. Here, we investigated the role of the putative mechanosensitive canonical transient receptor potential channel 1 (TRPC1) in cell migration. We analyzed its function in transformed renal epithelial (Madin–Darby canine kidney-focus) cells with variation of TRPC1 expression. As shown by time lapse video microscopy, TRPC1 knockdown cells have partially lost their polarity and the ability to persistently migrate into a given direction. This failure is linked to the suppression of a local Ca2+ gradient at the front of migrating TRPC1 knockdown cells, whereas TRPC1 overexpression leads to steeper Ca2+ gradients. We propose that the Ca2+ signaling events regulated by TRPC1 within the lamellipodium determine polarity and directed cell migration.

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Acknowledgments

The authors thank Michael Zhu from the Ohio State University for kindly providing us with a plasmid of the human HA-tagged TRPC1. This work was supported by a grant from the Rolf-Dierichs-Stiftung of the Medical Faculty of the University of Münster (grant number 193423) to A.F., by the IZKF Münster (grant number Schw2/030/08) and by the Deutsche Forschungsgemeinschaft (grant number Schw 407/9-3, 10-1) to A.S. B.N. was supported by a Human Frontiers Research Grant (RGP 32/2004), the Excellentiefinanciering (Flanders, EF/95/010), and the Interuniversity Poles of Attraction Program, Prime Minister’s Office IUAP.

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Authors and Affiliations

  1. Institute of Physiology II, University of Münster, Robert-Koch-Str. 27b, 48149, Münster, Germany

    A. Fabian, T. Fortmann, C. Riethmüller, P. Schön, S. Mally & A. Schwab

  2. Institute of Physiology, TU, Dresden, Germany

    P. Dieterich

  3. Life Science Team, Veeco Instruments GmbH, Mannheim, Germany

    P. Schön

  4. Department of Molecular Cell Biology, Lab Ion Channel Research, Campus Gasthuisberg, KU, Leuven, Belgium

    B. Nilius

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  1. A. Fabian
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  2. T. Fortmann
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Correspondence to A. Fabian.

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424_2008_515_MOESM1_ESM.mov

Video 1 Suppression of TRPC1 leads to loss of cellular polarity and impaired migration. Phase-contrast time lapse video microscopy of representative MDCK-F cells migrating on fibronectin for 3 h. Frames were collected in 1-min intervals and are displayed at a rate of 30 frames per second. Cells with knockdown of TRPC1 are shown in the left panel and hTRPC1-HA-overexpressing cells in the right panel. TRPC1 knockdown leads to a loss of cell polarity. A clear front-rear axis can no longer be recognized. Multiple broad lamellipodia are protruding into different directions simultaneously. In contrast, hTRPC1-HA-overexpressing cells are highly polarized and show a normal and efficient migration. As movies are shown in the same magnification, it is obvious that the projected cell surface of TRPC1-depleted cells has at least doubled. Scale bar: 50 µm. (MOV 2.78 MB)

Table S1 Nucleotide sequences of oligonucleotides used for RNA interference (DOC 37 KB).

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Fabian, A., Fortmann, T., Dieterich, P. et al. TRPC1 channels regulate directionality of migrating cells. Pflugers Arch - Eur J Physiol 457, 475–484 (2008). https://doi.org/10.1007/s00424-008-0515-4

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  • DOI: https://doi.org/10.1007/s00424-008-0515-4

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