In vitro cell stretching technology (IsoStretcher) as an approach to unravel Piezo1-mediated cardiac mechanotransduction
Introduction
Mechanosensory transduction is an ancient sensing mechanism involving mechanosensitive (MS) ion channels. These highly efficient biological force-sensing macromolecules are tightly coupled to the mechanics of biological cell membranes. Together with the cytoskeleton, integrins, G-protein-coupled receptors and membrane-bound enzymes, they represent major molecular mechanosensors tasked with transducing mechanical stimuli, exerted on the cell membrane, into electrochemical intracellular signals (Hamill and Martinac, 2001; Ingber, 2006). By operating on a millisecond time scale these channels play a central role in the physiology of touch, hearing and blood pressure regulation, for example (Martinac, 2004). Force-induced changes in these channels result in electrical signal transduction mediated via ion fluxes defined by channel conductive properties (conductance, ion selectivity) and electrochemical ion gradients. Studies of MS ion channels have been conducted for more than 30 years, including the first report of MS channels in bacteria (Martinac et al., 1987), which have greatly contributed to our understanding of the basic biophysical principles underlying the physiology of mechanosensory transduction in higher eukaryotes, including animals and humans (Cox et al., 2019; Martinac and Cox, 2017). However, a complete understanding of the molecular force-sensing mechanisms at play remains elusive.
The recent exciting discovery of the Piezo mechanically gated ion channel family (Coste et al., 2010) and their role in eukaryotes (Kim et al., 2012) has opened the possibility of closing the gap in our knowledge of the physiology and pathology of mechanotransduction processes. However, as we will point out, biological assessment of such MS entities requires availability of novel, organ-mimicking stretch technologies, which we will review alongside with our recently bioengineered IsoStretcher system technology that was here applied to study Piezo1 channel properties in murine atrial HL-1 cells.
Section snippets
Mechanosensitivity of Piezo1 channels
The Piezo channel family includes two isoforms: Piezo1 and Piezo2. Acting as Ca2+-permeable non-selective cation channels, they were initially indicated in 2010 to play important roles in mechanotransduction processes in various species including animal, plant and other eukaryotic species, throughout most tissues and cells (Coste et al., 2010). The two isoforms of Piezo encoded as PIEZO1 and PIEZO2 genes are sharing approximate 50% gene identity with each other and were considered to have some
Overview of stretching technologies
As the questions around mechanosensitive ion channels, such as Piezo1, have become increasingly important, many devices have been built to exert mechanical strain on cells. Various approaches emerged in the field of cell stretching, a few of which are presented and discussed here before introducing the IsoStretcher, an isotropic cell stretching device.
Furthermore, other groups have concentrated on alternative stimulation methods, like shear-stress, indentation and pressure. These methods were
Conclusion
MS channel Piezo1 is involved in a broad range of physiological and pathological processes in the body. In the cardiovascular system, the activities of Piezo1 have been mainly examined in ECs or RBCs. Importantly, Piezo1 is also thought to be able to respond to mechanical forces such as stretch due to cardiac contraction and to act as a primary force sensor in the heart, although the exact role of Piezo1 in cardiomyocytes still needs to be unravelled through further studies using different
Adult mouse cardiac fibroblast isolation and primary culture
Three 12-week-old male C57BL/6J mice with an average body weight of 29.53 g (SD = 0.81) were euthanized according to guidelines of the Animal Research Act 1985 No 123 (New South Wales, Australia). The isolation and primary culture of mouse CFs were following a published protocol by Zeigler et al. (2016).
Quantitative real-time polymerase chain reaction (RT-PCR)
RNA was extracted from adult mouse CFs and cultured HL-1 cells with the RNeasy Fibrous Tissue Mini Kit (QIAGEN), following the manufacturer’s protocol. cDNA was synthesized using the SuperScript
Author contributions
Yang Guo: Conceptualization, Data curation, Formal analysis, Methodology, Visualization, Writing - original draft, Writing - review & editing.
Anna-Lena Merten: Conceptualization, Data curation, Formal analysis, Methodology, Software, Visualization, Writing - original draft, Writing - review & editing.
Ulrike Schöler: Data curation, Formal analysis, Writing - review & editing.
Ze-Yan Yu: Supervision, Validation, Writing - review & editing
Jasmina Cvetkovska: Data curation.
Diane Fatkin: Data
Declaration of competing interest
The authors declare that they have no conflict of interest.
Acknowledgements
The authors are grateful for the support and advice of Dr. Charles D Cox in the HL-1 cell stretching experiments, as well as Dr. Louise Dunn and Professor Roland Stocker for their help in designing the mouse Piezo1 RT-PCR primers.
The authors gratefully acknowledge funding of the German Federal Ministry for Economy and Energy due to a resolution by the German Bundestag (ZIM and BMWi, #ZF4134304CR6) and funding of the Erlangen Graduate School in Advanced Optical Technologies (SAOT) by the German
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The role of Piezo1 and Piezo2 proteins in tissue engineering: A Comprehensive review
2024, Engineered RegenerationA narrative review on the biology of piezo1 with platelet-rich plasma in cardiac cell regeneration
2022, Chemico-Biological InteractionsCitation Excerpt :Surprisingly very little is documented about Piezo1's involvement in the Myocardium, which may result from its recent discovery. In this, Piezo1 was expressed in a range of cardiac cell types, including epithelial cells (EC), cardiac fibroblasts (CF), and cardiac myocytes (CM) in both human and animal samples, with a higher expression level in fibroblasts than myocytes [16,17] (Fig. 2). These observations may be used to calculate the rate of Ca2+ accumulation, the latency, and the time required to achieve the peak concentration.
Plasma membrane disruption (PMD) formation and repair in mechanosensitive tissues
2021, BoneCitation Excerpt :Mechanosensation mechanisms, or the processes by which cells detect and recognize mechanical stimuli, remain an area of rigorous study in many fields. Cells concurrently utilize myriad mechanisms to recognize mechanical stimuli, such as stretch-gated ion channels, chemical or force-mediated conformational changes of the cytoskeleton, molecular transport through channel proteins like gap junctions, hemichannels, and pannexins, and activation of voltage-gated calcium channels [7–20]. Another such mechanosensation mechanism is the loading-induced development of small, transient discontinuities in the cell membrane called plasma membrane disruptions (PMD) [21].
Mechanobiology of the cardiovascular system
2021, Progress in Biophysics and Molecular BiologyHigh-content method for mechanosignaling studies using IsoStretcher technology and quantitative Ca<sup>2+</sup> imaging applied to Piezo1 in cardiac HL-1 cells
2024, Cellular and Molecular Life SciencesPiezo1 in Digestive System Function and Dysfunction
2023, International Journal of Molecular Sciences
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These authors contributed equally.