| Autor: |
Capponi G; Dipartimento di Fisica e Geologia, Università di Perugia, 06100 Perugia, Italy.; Sezione di Farmacologia, Dipartimento di Medicina Interna, Università di Genova, 16132 Genova, Italy., Zambito M; Sezione di Farmacologia, Dipartimento di Medicina Interna, Università di Genova, 16132 Genova, Italy., Neri I; Dipartimento di Fisica e Geologia, Università di Perugia, 06100 Perugia, Italy., Cottone F; Dipartimento di Fisica e Geologia, Università di Perugia, 06100 Perugia, Italy., Mattarelli M; Dipartimento di Fisica e Geologia, Università di Perugia, 06100 Perugia, Italy., Vassalli M; James Watt School of Engineering, University of Glasgow, Glasgow G12 8LT, UK., Caponi S; Istituto Officina dei Materiali, Italian National Research Council (IOM-CNR), Unit of Perugia, c/o Department of Physics and Geology, University of Perugia, Via A. Pascoli, 06123 Perugia, Italy., Florio T; Sezione di Farmacologia, Dipartimento di Medicina Interna, Università di Genova, 16132 Genova, Italy.; Istituto di Ricovero e Cura a Carattere Scientifico, Ospedale Policlinico San Martino, 16132 Genova, Italy. |
| Abstrakt: |
Mechanotransduction refers to the cellular ability to sense mechanical stimuli from the surrounding environment and convert them into biochemical signals that regulate cellular physiology and homeostasis. Mechanosensitive ion channels (MSCs), especially ones of Piezo family (Piezo1 and Piezo2), play a crucial role in mechanotransduction. These transmembrane proteins directly react to mechanical cues by triggering the onset of an ionic current. The relevance of this mechanism in driving physiology and pathology is emerging, and there is a growing need for the identification of an affordable and reliable assay to measure it. Setting up a mechanosensitivity assay requires exerting a mechanical stimulus on single cells while observing the downstream effects of channels opening. We propose an open-hardware approach to stimulate single adherent cells through controlled microindentation, using a 3D-printed actuation platform. We validated the device by measuring the mechanosensitivity of a neural mice cell line where the expression level and activity of Piezo1 were genetically and pharmacologically manipulated. Moreover, this extremely versatile device could be integrated with different read-out technologies, offering a new tool to improve the understanding of mechanotransduction in living cells. |
