scholarly journals GenEPi: Piezo1-based fluorescent reporter for visualizing mechanical stimuli with high spatiotemporal resolution

2019 ◽  
Author(s):  
Sine Yaganoglu ◽  
Nordine Helassa ◽  
Benjamin M. Gaub ◽  
Maaike Welling ◽  
Jian Shi ◽  
...  
Keyword(s):  
Ion Channel ◽  
High Specificity ◽  
Mechanical Stimuli ◽  
Spatiotemporal Resolution ◽  
Fluorescent Reporter ◽  
Non Invasive ◽  
Live Tissue ◽  
Biological Responses ◽  
Mechanosensitive Ion Channel ◽  
Cellular Mechanosensing

AbstractMechanosensing is a ubiquitous process to translate external mechanical stimuli into biological responses during development, homeostasis, and disease. However, non-invasive investigation of cellular mechanosensing in complex and intact live tissue remains challenging. Here, we developed GenEPi, a genetically-encoded fluorescent intensiometric reporter for mechanical stimuli based on Piezo1, an essential mechanosensitive ion channel found in vertebrates. We show that GenEPi has high specificity and spatiotemporal resolution for Piezo1-dependent mechanical stimuli, exemplified by resolving repetitive mechanical stimuli of spontaneously contracting cardiomyocytes within microtissues, in a non-invasive manner.

scholarly journals TACAN is an essential component of the mechanosensitive ion channel responsible for pain sensing

2018 ◽  
Author(s):  
L. Beaulieu-Laroche ◽  
M. Christin ◽  
AM Donoghue ◽  
F. Agosti ◽  
N. Yousefpour ◽  
...  
Keyword(s):  
Ion Channels ◽  
Heterologous Expression ◽  
Ion Channel ◽  
Behavioral Responses ◽  
Mechanical Stimuli ◽  
Thermal Pain ◽  
Fundamental Process ◽  
Mechanosensitive Ion Channel ◽  
Pain Stimuli ◽  
Essential Subunit

SummaryMechanotransduction, the conversion of mechanical stimuli into electrical signals, is a fundamental process underlying several physiological functions such as touch and pain sensing, hearing and proprioception. This process is carried out by specialized mechanosensitive ion channels whose identities have been discovered for most functions except pain sensing. Here we report the identification of TACAN (Tmem120A), an essential subunit of the mechanosensitive ion channel responsible for sensing mechanical pain. TACAN is expressed in a subset of nociceptors, and its heterologous expression increases mechanically-evoked currents in cell lines. Purification and reconstitution of TACAN in synthetic lipids generates a functional ion channel. Finally, knocking down TACAN decreases the mechanosensitivity of nociceptors and reduces behavioral responses to mechanical but not to thermal pain stimuli, without affecting the sensitivity to touch stimuli. We propose that TACAN is a pore-forming subunit of the mechanosensitive ion channel responsible for sensing mechanical pain.

scholarly journals Mechanosensitive Ion Channel Piezo1 Regulates Myocyte Fusion during Skeletal Myogenesis

2020 ◽  
Author(s):  
Huascar Pedro Ortuste Quiroga ◽  
Shingo Yokoyama ◽  
Massimo Ganassi ◽  
Kodai Nakamura ◽  
Tomohiro Yamashita ◽  
...  
Keyword(s):  
Ion Channel ◽  
Molecular Mechanisms ◽  
Myoblast Fusion ◽  
Mechanical Stimuli ◽  
Muscle Satellite Cell ◽  
Myotube Formation ◽  
Mechanosensitive Ion Channel ◽  
And Function ◽  
Sirna Knockdown

AbstractMechanical stimuli such as stretch and resistance training are essential to regulate growth and function of skeletal muscle. However, the molecular mechanisms involved in sensing mechanical stress remain unclear. Here, the purpose of this study was to investigate the role of the mechanosensitive ion channel Piezo1 during myogenic progression. Muscle satellite cell-derived myoblasts and myotubes were modified with stretch, siRNA knockdown and agonist-induced activation of Piezo1. Direct manipulation of Piezo1 modulates terminal myogenic progression. Piezo1 knockdown suppressed myoblast fusion during myotube formation and maturation. This was accompanied by downregulation of the fusogenic protein Myomaker. Piezo1 knockdown also lowered Ca2+ influx in response to stretch. Conversely Piezo1 activation stimulated fusion and increased Ca2+ influx in response to stretch. These evidences indicate that Piezo1 is essential for myotube formation and maturation, which may have implications for msucular dystrophy prevention through its role as a mechanosensitive Ca2+ channel.

scholarly journals Acetylated tubulin is essential for touch sensation in mice

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Shane J Morley ◽  
Yanmei Qi ◽  
Loredana Iovino ◽  
Laura Andolfi ◽  
Da Guo ◽  
...  
Keyword(s):  
Ion Channel ◽  
Sensory Neurons ◽  
Mechanical Energy ◽  
Mechanical Stimuli ◽  
Acetylated Tubulin ◽  
Fundamental Level ◽  
Tubulin Acetylation ◽  
Channel Opening ◽  
Mechanosensitive Ion Channel ◽  
Touch Sensation

At its most fundamental level, touch sensation requires the translation of mechanical energy into mechanosensitive ion channel opening, thereby generating electro-chemical signals. Our understanding of this process, especially how the cytoskeleton influences it, remains unknown. Here we demonstrate that mice lacking the α-tubulin acetyltransferase Atat1 in sensory neurons display profound deficits in their ability to detect mechanical stimuli. We show that all cutaneous afferent subtypes, including nociceptors have strongly reduced mechanosensitivity upon Atat1 deletion, and that consequently, mice are largely insensitive to mechanical touch and pain. We establish that this broad loss of mechanosensitivity is dependent upon the acetyltransferase activity of Atat1, which when absent leads to a decrease in cellular elasticity. By mimicking α-tubulin acetylation genetically, we show both cellular rigidity and mechanosensitivity can be restored in Atat1 deficient sensory neurons. Hence, our results indicate that by influencing cellular stiffness, α-tubulin acetylation sets the force required for touch.

scholarly journals Fluorescence Microscopy of Piezo1 in Droplet Hydrogel Bilayers

2018 ◽  
Author(s):  
Oskar B. Jaggers ◽  
Pietro Ridone ◽  
Boris Martinac ◽  
Matthew A. B. Baker
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Simultaneous Recording ◽  
Mechanical Stimuli ◽  
Channel Activity ◽  
Channel Proteins ◽  
Mechanosensitive Ion Channel ◽  
Lymphatic Dysplasia ◽  
Hydrogel Bilayer

AbstractMechanosensitive ion channels are membrane gated pores which are activated by mechanical stimuli. The focus of this study is on Piezo1, a newly discovered, large, mammalian, mechanosensitive ion channel, which has been linked to diseases such as dehydrated hereditary stomatocytosis (Xerocytosis) and lymphatic dysplasia. Here we utilize an established in-vitro artificial bilayer system to interrogate single Piezo1 channel activity. The droplet-hydrogel bilayer (DHB) system uniquely allows the simultaneous recording of electrical activity and fluorescence imaging of labelled protein. We successfully reconstituted fluorescently labelled Piezo1 ion channels in DHBs and verified activity using electrophysiology in the same system. We demonstrate successful insertion and activation of hPiezo1-GFP in bilayers of varying composition. Furthermore, we compare the Piezo1 bilayer reconstitution with measurements of insertion and activation of KcsA channels to reproduce the channel conductances reported in the literature. Together, our results showcase the use of DHBs for future experiments allowing simultaneous measurements of ion channel gating while visualising the channel proteins using fluorescence.

scholarly journals Chondroprotection by urocortin involves blockade of the mechanosensitive ion channel Piezo1

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
K. M. Lawrence ◽  
R. C. Jones ◽  
T. R. Jackson ◽  
R. L. Baylie ◽  
B. Abbott ◽  
...  
Keyword(s):  
Ion Channel ◽  
Mechanosensitive Ion Channel

scholarly journals Non-Invasive Molecular Survey of Sarcoptic Mange in Wildlife: Diagnostic Performance in Wolf Faecal Samples Evaluated by Multi-Event Capture–Recapture Models

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 243
Author(s):  
Julieta Rousseau ◽  
Mónia Nakamura ◽  
Helena Rio-Maior ◽  
Francisco Álvares ◽  
Rémi Choquet ◽  
...  
Keyword(s):  
Molecular Detection ◽  
High Specificity ◽  
Sarcoptes Scabiei ◽  
Sarcoptic Mange ◽  
Diagnostic Specificity ◽  
Non Invasive ◽  
Faecal Samples ◽  
Capture Recapture ◽  
The Mean ◽  
Event Capture

Sarcoptic mange is globally enzootic, and non-invasive methods with high diagnostic specificity for its surveillance in wildlife are lacking. We describe the molecular detection of Sarcoptes scabiei in non-invasively collected faecal samples, targeting the 16S rDNA gene. We applied this method to 843 Iberian wolf Canis lupus signatus faecal samples collected in north-western Portugal (2006–2018). We further integrated this with serological data (61 samples from wolf and 20 from red fox Vulpes vulpes, 1997–2019) in multi-event capture–recapture models. The mean predicted prevalence by the molecular analysis of wolf faecal samples from 2006–2018 was 7.2% (CI95 5.0–9.4%; range: 2.6–11.7%), highest in 2009. The mean predicted seroprevalence in wolves was 24.5% (CI95 18.5–30.6%; range: 13.0–55.0%), peaking in 2006–2009. Multi-event capture–recapture models estimated 100% diagnostic specificity and moderate diagnostic sensitivity (30.0%, CI95 14.0–53.0%) for the molecular method. Mange-infected individually identified wolves showed a tendency for higher mortality versus uninfected wolves (ΔMortality 0.150, CI95 −0.165–0.458). Long-term serology data highlights the endemicity of sarcoptic mange in wild canids but uncovers multi-year epidemics. This study developed and evaluated a novel method for surveying sarcoptic mange in wildlife populations by the molecular detection of S. scabiei in faecal samples, which stands out for its high specificity and non-invasive character.

scholarly journals Identification of Tumor-Specific MRI Biomarkers Using Machine Learning (ML)

Diagnostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 742
Author(s):  
Rima Hajjo ◽  
Dima A. Sabbah ◽  
Sanaa K. Bardaweel ◽  
Alexander Tropsha
Keyword(s):  
Machine Learning ◽  
High Specificity ◽  
Cancer Biomarkers ◽  
Cancer Prognosis ◽  
Current Status ◽  
Imaging Data ◽  
Non Invasive ◽  
Machine Learning Methods ◽  
Cancer Types ◽  
Magnetic Resonance Imaging Mri

The identification of reliable and non-invasive oncology biomarkers remains a main priority in healthcare. There are only a few biomarkers that have been approved as diagnostic for cancer. The most frequently used cancer biomarkers are derived from either biological materials or imaging data. Most cancer biomarkers suffer from a lack of high specificity. However, the latest advancements in machine learning (ML) and artificial intelligence (AI) have enabled the identification of highly predictive, disease-specific biomarkers. Such biomarkers can be used to diagnose cancer patients, to predict cancer prognosis, or even to predict treatment efficacy. Herein, we provide a summary of the current status of developing and applying Magnetic resonance imaging (MRI) biomarkers in cancer care. We focus on all aspects of MRI biomarkers, starting from MRI data collection, preprocessing and machine learning methods, and ending with summarizing the types of existing biomarkers and their clinical applications in different cancer types.

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