Disruption of membrane cholesterol organization impairs the activity of PIEZO1 channel clusters

Martinac, Boris and Gottlieb, Philip A. and Macmillan, Alexander and Cox, Charles D. and Vassalli, Massimo and Pandzic, Elvis and Ridone, Pietro (2020) Disruption of membrane cholesterol organization impairs the activity of PIEZO1 channel clusters. Journal of General Physiology, 152 (8). ISSN 0022-1295

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Link to published document: http://doi.org/10.1085/jgp.201912515

Abstract

The human mechanosensitive ion channel PIEZO1 is gated by membrane tension and regulates essential biological processes such as vascular development and erythrocyte volume homeostasis. Currently, little is known about PIEZO1 plasma membrane localization and organization. Using a PIEZO1-GFP fusion protein, we investigated whether cholesterol enrichment or depletion by methyl-β-cyclodextrin (MBCD) and disruption of membrane cholesterol organization by dynasore affects PIEZO1-GFP's response to mechanical force. Electrophysiological recordings in the cell-attached configuration revealed that MBCD caused a rightward shift in the PIEZO1-GFP pressure-response curve, increased channel latency in response to mechanical stimuli, and markedly slowed channel inactivation. The same effects were seen in native PIEZO1 in N2A cells. STORM superresolution imaging revealed that, at the nanoscale, PIEZO1-GFP channels in the membrane associate as clusters sensitive to membrane manipulation. Both cluster distribution and diffusion rates were affected by treatment with MBCD (5 mM). Supplementation of polyunsaturated fatty acids appeared to sensitize the PIEZO1-GFP response to applied pressure. Together, our results indicate that PIEZO1 function is directly dependent on the membrane composition and lateral organization of membrane cholesterol domains, which coordinate the activity of clustered PIEZO1 channels.

Item Type: Article
Subjects: R Medicine > R Medicine (General)
Divisions: Faculty of Medicine, Health and Life Sciences > School of Medicine
Depositing User: Repository Administrator
Date Deposited: 02 Jul 2020 03:50
Last Modified: 19 Oct 2021 01:11
URI: http://eprints.victorchang.edu.au/id/eprint/990

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