Sphingomyelinase Disables Inactivation in Endogenous PIEZO1 Channels
Jian Shi,
Adam J. Hyman,
Dario De Vecchis,
Jiehan Chong,
Laeticia Lichtenstein,
T. Simon Futers,
Myriam Rouahi,
Anne Negre Salvayre,
Nathalie Auge,
Antreas C. Kalli,
David J. Beech
Affiliations
Jian Shi
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK; Corresponding author
Adam J. Hyman
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK
Dario De Vecchis
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK
Jiehan Chong
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK
Laeticia Lichtenstein
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK
T. Simon Futers
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK
Myriam Rouahi
INSERM U-1048 and Université Paul Sabatier, 31432 Cedex 4 Toulouse, France
Anne Negre Salvayre
INSERM U-1048 and Université Paul Sabatier, 31432 Cedex 4 Toulouse, France
Nathalie Auge
INSERM U-1048 and Université Paul Sabatier, 31432 Cedex 4 Toulouse, France
Antreas C. Kalli
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK; Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK; Corresponding author
David J. Beech
Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK; Corresponding author
Summary: Endogenous PIEZO1 channels of native endothelium lack the hallmark inactivation often seen when these channels are overexpressed in cell lines. Because prior work showed that the force of shear stress activates sphingomyelinase in endothelium, we considered if sphingomyelinase is relevant to endogenous PIEZO1. Patch clamping was used to quantify PIEZO1-mediated signals in freshly isolated murine endothelium exposed to the mechanical forces caused by shear stress and membrane stretch. Neutral sphingomyelinase inhibitors and genetic disruption of sphingomyelin phosphodiesterase 3 (SMPD3) cause PIEZO1 to switch to profoundly inactivating behavior. Ceramide (a key product of SMPD3) rescues non-inactivating channel behavior. Its co-product, phosphoryl choline, has no effect. In contrast to ceramide, sphingomyelin (the SMPD3 substrate) does not affect inactivation but alters channel force sensitivity. The data suggest that sphingomyelinase activity, ceramide, and sphingomyelin are determinants of native PIEZO gating that enable sustained activity.
