title: Mechanosensitive ion channel Piezo2 is inhibited by D-GsMTx4.
abstract: Enterochromaffin (EC) cells are the primary mechanosensors of the gastrointestinal (GI) epithelium. In response to mechanical stimuliECcells release serotonin (5-hydroxytryptamine; 5-HT). The molecular details ofECcell mechanosensitivity are poorly understood. Recently, our group found that human and mouseECcells express the mechanosensitive ion channel Piezo2. The mechanosensitive currents in a humanECcell model QGP-1 were blocked by the mechanosensitive channel blocker D-GsMTx4. In the present study we aimed to characterize the effects of the mechanosensitive ion channel inhibitor spider peptide D-GsMTx4 on the mechanically stimulated currents from both QGP-1 and human Piezo2 transfected HEK-293 cells. We found co-localization of 5-HT and Piezo2 in QGP-1 cells by immunohistochemistry. QGP-1 mechanosensitive currents had biophysical properties similar to dose-dependently Piezo2 and were inhibited by D-GsMTx4. In response to direct displacement of cell membranes, human Piezo2 transiently expressed in HEK-293 cells produced robust rapidly activating and inactivating inward currents. D-GsMTx4 reversibly and dose-dependently inhibited both the potency and efficacy of Piezo2 currents in response to mechanical force. Our data demonstrate an effective inhibition of Piezo2 mechanosensitive currents by the spider peptide D-GsMTx4.
authors.affiliation: a Department of Physiology & Biomedical Engineering, Enteric Neuroscience Program, Division of Gastroenterology & Hepatology , Mayo Clinic , Rochester , MN , USA.
authors.forename: Constanza
authors.initials: C
authors.lastname: Alcaino
authors.affiliation: a Department of Physiology & Biomedical Engineering, Enteric Neuroscience Program, Division of Gastroenterology & Hepatology , Mayo Clinic , Rochester , MN , USA.
authors.forename: Kaitlyn
authors.initials: K
authors.lastname: Knutson
authors.affiliation: b Department of Physiology and Biophysics, Center for Single Molecule Biophysics , State University of New York , Buffalo , NY , USA.
authors.forename: Philip A
authors.initials: PA
authors.lastname: Gottlieb
authors.affiliation: b Department of Physiology and Biophysics, Center for Single Molecule Biophysics , State University of New York , Buffalo , NY , USA.
authors.forename: Gianrico
authors.initials: G
authors.lastname: Farrugia
authors.affiliation: a Department of Physiology & Biomedical Engineering, Enteric Neuroscience Program, Division of Gastroenterology & Hepatology , Mayo Clinic , Rochester , MN , USA.
authors.forename: Arthur
authors.initials: A
authors.lastname: Beyder
mesh_terms: D001696:Biomechanical Phenomena; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007473:Ion Channels / Q000037:antagonists & inhibitors* / Q000378:metabolism; D040542:Mechanotransduction, Cellular / Q000187:drug effects; D010455:Peptides / Q000494:pharmacology*; D013111:Spider Venoms / Q000494:pharmacology*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D007473:Ion Channels; C411249:MTx4 protein, Grammostola spatulata; C000589132:PIEZO2 protein, human; D010455:Peptides; D013111:Spider Venoms
keywords: D-GsMTx4; Piezo2; enterochromaffin cell; mechanosensitive ion channel; mechanotransduction