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Mechanically-Evoked TRPV4-Mediated Currents Are Modulated by Activated Integrin β1

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Mechanobiology (ISMB 2022)

Part of the book series: Springer Series in Biophysics ((BIOPHYSICS,volume 25))

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Abstract.

TRPV4 is a polymodal, non-selective cation channel which can be activated by mechanical stimuli only when they are applied directly at the cell-substrate interface. This restricted profile of mechanical activation suggests that mechanical gating of TRPV4 is highly dependent on either the specific physical context or molecular interaction partners within this cellular compartment. However, it is presently unknown what aspects of the cell-substrate interface confer mechanosensitivity to TRPV4. Integrin β1 has previously been indirectly linked to the mechanical activation of TRPV4 but this link has not yet been investigated in assays where it is clear that the gating stimulus was the mechanical input. We therefore aimed to test whether increasing integrin β1 activation could sensitise TRPV4-mediated, mechanically-evoked currents using whole-cell patch-clamp electrophysiology combined with a technique to apply mechanical stimuli directly at cell-substrate connections. We found that TRPV4-mediated currents were sensitised when integrin β1 was activated using the functional anti-integrin β1 [12G10] antibody. However, activation of integrin β1 using fibronectin-coated substrates only lead to minor increases in TRPV4 sensitivity. These data suggest that mechanical signalling via TRPV4 in the cell-substrate interface can be tuned by the activation state of integrin β1.

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Acknowledgements

This work supported by a NHMRC project grant to KP (APP1122104) and an Australian Government RTP and UNSW School of Medical Sciences funded scholarship to JR. The HEK-293T PIEZO1 knockout cell line was kindly gifted by Ardem Patapoutian. Confocal imaging was conducted in the Katharina Gaus Light Microscopy Facility (KGLMF), at the UNSW Mark Wainwright Analytical Centre. The authors also acknowledge the traditional custodians of the land on which this work was conducted, the Bidjigal people of the Eora nation, and the authors pay their respects to Elders past, present, and emerging.

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Authors and Affiliations

  1. School of Biomedical Sciences, Faculty of Medicine and Health, University of New South Wales, Sydney, NSW, Australia

    Jessica Richardson, Lioba Schroeter & Kate Poole

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  1. Jessica Richardson
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  2. Lioba Schroeter
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  3. Kate Poole
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Correspondence to Kate Poole .

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Editors and Affiliations

  1. Lowry Packer Building, Victor Chang Cardiac Research Institute, Darlinghurst, NSW, Australia

    Boris Martinac

  2. Lowy Packer Building, Level 6, Victor Chang Cardiac Research Institute, Darlinghurst, NSW, Australia

    Charles D. Cox

  3. School of Biomedical Sciences, UNSW Sydney, Sydney, NSW, Australia

    Kate Poole

  4. Baker Heart and Diabetes Institute, Melbourne, VIC, Australia

    Sara Baratchi

  5. EMBL Australia, Single Molecule Science node, School of Biomedical Sciences, UNSW Sydney, Sydney, NSW, Australia

    Daryan Kempe

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Richardson, J., Schroeter, L., Poole, K. (2024). Mechanically-Evoked TRPV4-Mediated Currents Are Modulated by Activated Integrin β1. In: Martinac, B., Cox, C.D., Poole, K., Baratchi, S., Kempe, D. (eds) Mechanobiology. ISMB 2022. Springer Series in Biophysics, vol 25. Springer, Cham. https://doi.org/10.1007/978-3-031-45379-3_1

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