Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi

Yaganoglu, Sine; Kalyviotis, Konstantinos; Vagena-Pantoula, Christina; Jülich, Dörthe; Gaub, Benjamin M.; Welling, Maaike; Lopes, Tatiana; Lachowski, Dariusz; Tang, See Swee; Hernández, Armando E. del Río; Salem, Victoria; Müller, Daniel J.; Holley, Scott A.; Vermot, Julien; Shi, Jing; Helassa, Nordine; Török, Katalin; Pantazis, Periklis

doi:10.1038/s41467-023-40134-y

Cited by 9 publications

(6 citation statements)

References 89 publications

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“…These significant technical advancements over previous imaging-based approaches to measure PIEZO1 activity 26–28,30,32 , demonstrate our tool to be exceptionally suited for examining both PIEZO1 localization and activity dynamics under native cellular conditions in isolated cells as well as tissue organoids. Our findings contrast with existing methodologies, such as GenEPi 53 , a genetically-encoded fluorescent reporter of PIEZO1 activity, which relies on overexpression and is limited by poor kinetics and weaker signals. In comparison, the PIEZO1-HaloTag hiPSC circumvents these pitfalls and provides a physiologically-relevant representation of PIEZO1 behavior.…”

Section: Discussioncontrasting

confidence: 88%

Visualizing PIEZO1 Localization and Activity in hiPSC-Derived Single Cells and Organoids with HaloTag Technology

Bertaccini,

Casanellas,

Evans

et al. 2023

Preprint

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PIEZO1 channels play a critical role in numerous physiological processes by transducing diverse mechanical stimuli into electrical and chemical signals. Recent studies underscore the importance of endogenous PIEZO1 activity and localization in regulating mechanotransduction. To enable physiologically and clinically relevant human-based studies, we genetically engineered human induced pluripotent stem cells (hiPSCs) to express a HaloTag fused to endogenous PIEZO1. Combined with super-resolution imaging, our chemogenetic approach allows precise visualization of PIEZO1 in various cell types. Further, the PIEZO1-HaloTag hiPSC technology allows non-invasive monitoring of channel activity via Ca2+-sensitive HaloTag ligands, with temporal resolution approaching that of patch clamp electrophysiology. Using lightsheet imaging of hiPSC-derived neural organoids, we also achieve molecular scale PIEZO1 imaging in three-dimensional tissue samples. Our advances offer a novel platform for studying PIEZO1 mechano-transduction in human cells and tissues, with potential for elucidating disease mechanisms and development of targeted therapeutics.

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Section: Discussioncontrasting

confidence: 88%

Visualizing PIEZO1 Localization and Activity in hiPSC-Derived Single Cells and Organoids with HaloTag Technology

Bertaccini,

Casanellas,

Evans

et al. 2023

Preprint

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“…Our findings provide insights into the importance of understanding mechanical forces on the cellular level during development. Recent work has brought the ability to visualize Piezo1 activity into fruition using GenEPi [ 60 ]. Using this new toolset would provide great insight into the roles of Piezo, while identifying when and where it is active.…”

Section: Discussionmentioning

confidence: 99%

Piezo1-mediated spontaneous calcium transients in satellite glia impact dorsal root ganglia development

Brandt,

Smith

2023

PLoS Biol

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Spontaneous Ca2+ transients of neural cells is a hallmark of the developing nervous system. It is widely accepted that chemical signals, like neurotransmitters, contribute to spontaneous Ca2+ transients in the nervous system. Here, we reveal an additional mechanism of spontaneous Ca2+ transients that is mechanosensitive in the peripheral nervous system (PNS) using intravital imaging of growing dorsal root ganglia (DRG) in zebrafish embryos. GCaMP6s imaging shows that developing DRG satellite glia contain distinct spontaneous Ca2+ transients, classified into simultaneous, isolated, and microdomains. Longitudinal analysis over days in development demonstrates that as DRG satellite glia become more synchronized, isolated Ca2+ transients remain constant. Using a chemical screen, we identify that Ca2+ transients in DRG glia are dependent on mechanical properties, which we confirmed using an experimental application of mechanical force. We find that isolated spontaneous Ca2+ transients of the glia during development is altered by manipulation of mechanosensitive protein Piezo1, which is expressed in the developing ganglia. In contrast, simultaneous Ca2+ transients of DRG satellite glia is not Piezo1-mediated, thus demonstrating that distinct mechanisms mediate subtypes of spontaneous Ca2+ transients. Activating Piezo1 eventually impacts the cell abundance of DRG cells and behaviors that are driven by DRG neurons. Together, our results reveal mechanistically distinct subtypes of Ca2+ transients in satellite glia and introduce mechanobiology as a critical component of spontaneous Ca2+ transients in the developing PNS.

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“… 77 Given that MSCs like Piezo1 also participate in many complex processes that is usually transient and could not be interrogated through current strategies, such as learning and memory, in which the real‐time recording of MSC‐dependent neuronal activities is necessary. 78 Using strategies such as genetically encoded fluorescent reporter of piezo1, 79 as well as the in vivo microscopic or mesoscopic imaging, it might be helpful to study the complicated MSC functions in more depth. Developing novel fluorescent reporters for optical recording MSC activity would be also helpful.…”

Section: Functional Roles Of Mscs In the Cerebral Neurophysiological ...mentioning

confidence: 99%

Mechanobiological insight into brain diseases based on mechanosensitive channels: Common mechanisms and clinical potential

Li,

Zhao,

Tian

et al. 2024

CNS Neurosci Ther

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BackgroundAs physical signals, mechanical cues regulate the neural cells in the brain. The mechanosensitive channels (MSCs) perceive the mechanical cues and transduce them by permeating specific ions or molecules across the plasma membrane, and finally trigger a series of intracellular bioelectrical and biochemical signals. Emerging evidence supports that wide‐distributed, high‐expressed MSCs like Piezo1 play important roles in several neurophysiological processes and neurological disorders.AimsTo systematically conclude the functions of MSCs in the brain and provide a novel mechanobiological perspective for brain diseases.MethodWe summarized the mechanical cues and MSCs detected in the brain and the research progress on the functional roles of MSCs in physiological conditions. We then concluded the pathological activation and downstream pathways triggered by MSCs in two categories of brain diseases, neurodegenerative diseases and place‐occupying damages. Finally, we outlined the methods for manipulating MSCs and discussed their medical potential with some crucial outstanding issues.ResultsThe MSCs present underlying common mechanisms in different brain diseases by acting as the “transportation hubs” to transduce the distinct signal patterns: the upstream mechanical cues and the downstream intracellular pathways. Manipulating the MSCs is feasible to alter the complicated downstream processes, providing them promising targets for clinical treatment.ConclusionsRecent research on MSCs provides a novel insight into brain diseases. The common mechanisms mediated by MSCs inspire a wide range of therapeutic potentials targeted on MSCs in different brain diseases.

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Highly specific and non-invasive imaging of Piezo1-dependent activity across scales using GenEPi

Cited by 9 publications

References 89 publications

Visualizing PIEZO1 Localization and Activity in hiPSC-Derived Single Cells and Organoids with HaloTag Technology

Visualizing PIEZO1 Localization and Activity in hiPSC-Derived Single Cells and Organoids with HaloTag Technology

Piezo1-mediated spontaneous calcium transients in satellite glia impact dorsal root ganglia development

Mechanobiological insight into brain diseases based on mechanosensitive channels: Common mechanisms and clinical potential

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