Pressure sensing through Piezo channels controls whether cells migrate with blebs or pseudopods

Srivastava, Nishit; Traynor, David; Piel, Matthieu; Kabla, Alexandre; Kay, Robert R.

doi:10.1073/pnas.1905730117

Cited by 64 publications

(74 citation statements)

References 67 publications

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“…Different microfluidic systems were recently designed to determine the impact of confinement on single [21][22][23][24] and collective 25,26 cell migration, nucleus deformation, [27][28][29][30][31][32] rupture and repair 33,34 and cell division. 35,36 Using such systems, a switch from a mesenchymal to an amoeboid mode of migration upon cell confinement was highlighted for various mesenchymal cell types 37 including embryonic progenitor cells.…”

Section: Introductionmentioning

confidence: 99%

A new agarose-based microsystem to investigate cell response to prolonged confinement

et al. 2020

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Section: Introductionmentioning

confidence: 99%

A new agarose-based microsystem to investigate cell response to prolonged confinement

et al. 2020

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“…Recently, Srivastava et al identified Piezo in Dictyostelium cells as a pressure sensor, required for a Ca 2+ -dependent switch from pseudopod to bleb-based migration 49 . We similarly observed that compressing cells beyond the point of initial contact with the pillar resulted in elevated cytosolic Ca 2+ and induced blebbing.…”

Section: Similarly Changes In Actomyosin Contractility and Substramentioning

confidence: 99%

Piezo1 activation attenuates thrombin-induced blebbing in breast cancer cells

O’Callaghan

Engberg

Fatsis-Kavalopoulos

et al. 2020

Preprint

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Cancer cells exploit a variety of migration modes to leave primary tumors and establish metastases, including amoeboid cell migration facilitated by bleb formation. Here we demonstrate that thrombin induces dynamic blebbing in the MDA-MB-231 breast cancer cell line, and confirm that PAR2 activation is sufficient to induce this effect. Cell confinement has been implicated as a driving force in bleb-based migration. Unexpectedly, we find that gentle contact compression, exerted using a "Cell Press" to mechanically stimulate cells, attenuated thrombin-induced blebbing with an associated increase in cytosolic calcium. Thrombin-induced blebbing was similarly attenuated using Yoda1, an agonist of the mechanosensitive calcium channel Piezo1, and its capacity to attenuate blebbing was impaired in Piezo1 depleted cells. Additionally, Piezo1 activation suppressed and reversed the thrombininduced phosphorylation of ERM proteins, which are implicated in the blebbing process, and this activity was in part mediated through activation of the PP1A/PP2A family of serine/threonine phosphatases. Our results provide mechanistic insights into Piezo1 activation as a suppressor of dynamic blebbing, specifically that which is induced by thrombin.2 increase hydrostatic pressure within the cell 21 ; in parallel, actomyosin contractility is essential for the retraction of expanded PM blebs 5 .Piezo1 is a mechanosensitive cation channel 22,23 implicated in diverse areas of cell biology including shear stress sensing in endothelial cells 24 ; regulation of red blood cell volume 25 ; cell division in epithelial cells 26 ; and as a confinement sensor that suppresses PKA activity to optimize cell motility 27 . Piezo1 is found in the plasma membrane as a homotrimer and in closed conformation induces a bowlshaped indentation in the PM. In response to increased membrane tension Piezo1 transitions to a more planar arrangement and gates Ca 2+ influx 28,29 . Furthermore, loss of cortical actin, as seen during the initial expansion of PM blebs, reduces the activation threshold of Piezo1 30 .Here, we identify thrombin as a bleb induction stimulus, and Piezo1 activation, through contact compression or stimulation with the Piezo1 agonist Yoda1, as an effective mechanism to suppress dynamic blebbing in breast cancer cells. Thrombin induced blebbing was associated with increased ERM phosphorylation that was suppressed or reversed through Piezo1 activation, which in turn was dependent on the downstream activity of PP1/PP2A serine/threonine phosphatases. We propose that Ca 2+ influx via Piezo1 activates PP1/PP2A phosphatases to dephosphorylate ERMs, which in turn reduces the actomyosin contraction forces applied to the PM; thereby reducing the likelihood of dynamic blebbing. Materials and Methods Reagents and chemicalsSiR-Actin (Spirochrome, Tebu-bio, Roskilde, Denmark) was diluted to 1 mM in dimethyl sulfoxide (DMSO; ThermoFisher Scientific, Uppsala, Sweden) and stored at -20 o C. Fluo-4 and Fura Red calcium indicators (ThermoFisher Scientific) were resuspe...

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“…Moreover, mechanisms of motility vary even among amoebozoans. For example, while E. histolytica blebs (Maugis et al, 2010), D. discoideum can switch between blebbing and using actin-filled pseudopods for α-motility (Srivastava et al, 2020;Tyson et al, 2014;Zatulovskiy et al, 2014). Therefore, empirically determining the mechanisms that control phenotypes like motility and phagocytosis in an understudied organism is an important step toward understanding its cell biology.…”

Section: Introductionmentioning

confidence: 99%

Arp2/3 complex-mediated actin assembly drives microtubule-independent motility and phagocytosis in the evolutionarily divergent amoebaNaegleria

Fritz‐Laylin

2020

Preprint

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Much of our current understanding of actin-driven phenotypes in eukaryotes has come from the "yeast to human" opisthokont lineage, as well as the related amoebozoa. Outside of these groups lies the genus Naegleria, which diverged from humans over a billion years ago, and includes the deadly "braineating amoeba." Unlike nearly every other known eukaryotic cell type, Naegleria amoebae are thought to lack cytoplasmic microtubules. The absence of microtubules suggests that these amoebae rapidly crawl and phagocytose bacteria using actin alone. Although this makes Naegleria a powerful system to probe actin-driven functions in the absence of microtubules, surprisingly little is known about Naegleria's actin cytoskeleton. Here, we use microscopy and genomic analysis to show that Naegleria amoebae have an extensive actin cytoskeletal repertoire, complete with nucleators and nucleation promoting factors. Naegleria use this cytoskeletal machinery to generate Arp2/3-dependent lamellar protrusions, which correlate with the capacity to migrate and phagocytose bacteria. Because human cells also use Arp2/3-dependent lamellar protrusions for motility and phagocytosis, this work supports an evolutionarily ancient origin for these actin-driven processes and establishes Naegleria as a natural model system for studying microtubule-independent cytoskeletal phenotypes.

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Pressure sensing through Piezo channels controls whether cells migrate with blebs or pseudopods

Cited by 64 publications

References 67 publications

A new agarose-based microsystem to investigate cell response to prolonged confinement

A new agarose-based microsystem to investigate cell response to prolonged confinement

Piezo1 activation attenuates thrombin-induced blebbing in breast cancer cells

Arp2/3 complex-mediated actin assembly drives microtubule-independent motility and phagocytosis in the evolutionarily divergent amoebaNaegleria

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