Characterisation of Asp669Tyr Piezo1 cation channel activity in red blood cells: an unexpected phenotype

Pérès, Laurent; Alonso, David Monedero; Nudel, Morgane; Figeac, Martin; Bruge, Judith; Picard, Véronique; Nemer, Wassim El; Preudhomme, Claude; Rosé, Christian; Egée, Stéphane; Bouyer, Guillaume

doi:10.1111/bjh.17467

Cited by 7 publications

(11 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Data presented here show that the Na + and K + permeability of patients' RBC was increased in the three cases of PIEZO1 mutations (V598M, F681S, G782S/R808Q) and these RBC are slightly dehydrated due to chronic activation of KCNN4, as shown in our previous publication (Rapetti-Mauss et al, 2017). The properties of these RBC are similar to the ones recently described for the D669Y PIEZO1 mutant (Pérès et al, 2021). The D669Y PIEZO1 RBC have an increased cation conductance and slight dehydration likely due to more frequent activation of KCNN4.…”

Section: Discussionsupporting

confidence: 88%

“…In DHSt patients with PIEZO1 gain of function mutations, RBC are dehydrated and show a typical leftward shift in osmotic gradient ektacytometry (i.e., toward increased deformability at lower osmolality values), attesting changes in the water content equilibrium ( Andolfo et al, 2013 ; Picard et al, 2019 ; Pérès et al, 2021 ). The increasing use of sequencing in diagnosis has resulted in identification of a collection of mutations in PIEZO1 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Hereditary Xerocytosis: Differential Behavior of PIEZO1 Mutations in the N-Terminal Extracellular Domain Between Red Blood Cells and HEK Cells

et al. 2021

Self Cite

View full text Add to dashboard Cite

Hereditary Xerocytosis, a rare hemolytic anemia, is due to gain of function mutations in PIEZO1, a non-selective cation channel activated by mechanical stress. How these PIEZO1 mutations impair channel function and alter red blood cell (RBC) physiology, is not completely understood. Here, we report the characterization of mutations in the N-terminal part of the protein (V598M, F681S and the double mutation G782S/R808Q), a part of the channel that was subject of many investigations to decipher its role in channel gating. Our data show that the electrophysiological features of these PIEZO1 mutants expressed in HEK293T cells are different from previously characterized PIEZO1 mutations that are located in the pore or at the C-terminal extracellular domain of the protein. Although RBC with PIEZO1 mutations showed a dehydrated phenotype, the activity of V598M, F681S or R808Q in response to stretch was not significantly different from the WT channels. In contrast, the G782S mutant showed larger currents compared to the WT PIEZO1. Interestingly, basal activity of all the mutated channels was not significantly altered at the opposite of what was expected according to the decreased water and cation contents of resting RBC. In addition, the features of mutant PIEZO1 expressed in HEK293 cells do not always correlate with the observation in RBC where PIEZO1 mutations induced a cation leak associated with an increased conductance. Our work emphasizes the role of the membrane environment in PIEZO1 activity and the need to characterize RBC permeability to assess pathogenicity to PIEZO1 mutants associated with erythrocyte diseases.

show abstract

Section: Discussionsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Hereditary Xerocytosis: Differential Behavior of PIEZO1 Mutations in the N-Terminal Extracellular Domain Between Red Blood Cells and HEK Cells

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…Indeed, it was shown that mechanical force causes calcium influx into RBCs that is dependent on Piezo1 expression (Cahalan et al, 2015). Such activation causes Ca 2+ influx and eventually Gárdos channel-dependent RBC dehydration, with certainty the scenario leading to hereditary xerocytosis with many Piezo1 mutants described so far (Zarychanski et al, 2012;Cahalan et al, 2015;Andolfo et al, 2018;Rotordam et al, 2019;Peres et al, 2021). But for healthy RBCs, such transient activation (a few ms), will offer (i) a simple opportunity to adjust the cell volume without altering shape or deformability of the erythrocyte in capillaries below their own volume, (ii) back to the normal bloodstream, activation of a NSC will tend to let Na + entering the cell, allowing near full restoration of cell volume, (iii) which eventually should give enough time for the Na + /K + pump (considering the rate of exchange) to finely tune the intracellular Na + /K + ratio to maintain homeostasis.…”

Section: Discussionmentioning

confidence: 99%

“…The CCCP method (Macey et al, 1978;Bennekou and Christophersen, 1986;Peres et al, 2021) or MBE method as renamed recently (Jansen et al, 2021) was used for the monitoring of membrane potential evolution. When erythrocytes are suspended in nominally buffer-free solution in the presence of the protonophore CCCP, changes in extracellular pH reflect membrane potential changes, since protons are kept at equilibrium across the membrane.…”

Section: Membrane Potential Estimationmentioning

confidence: 99%

“…Measuring membrane potential changes via the CCCP (carbonyl cyanide-m-chloro-phenyl-hydrazone) method has proven to be a valuable tool in the study of NSC activity (Macey et al, 1978;Bennekou et al, 2006;Filser et al, 2021;Peres et al, 2021). For instance, inhibition of G Cl -in Low Ionic Strength conditions (LIS) revealed NSC channel activity (Bennekou et al, 2003) which are the correlate of NSVDC channels previously described by patch-clamp at the single level (Christophersen and Bennekou, 1991;Kaestner et al, 1999Kaestner et al, , 2000.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The Chloride Conductance Inhibitor NS3623 Enhances the Activity of a Non-selective Cation Channel in Hyperpolarizing Conditions

et al. 2021

Self Cite

View full text Add to dashboard Cite

Handbooks of physiology state that the strategy adopted by red blood cells (RBCs) to preserve cell volume is to maintain membrane permeability for cations at its minimum. However, enhanced cation permeability can be measured and observed in specific physiological and pathophysiological situations such as in vivo senescence, storage at low temperature, sickle cell anemia and many other genetic defects affecting transporters, membrane or cytoskeletal proteins. Among cation pathways, cation channels are able to dissipate rapidly the gradients that are built and maintained by the sodium and calcium pumps. These situations are very well-documented but a mechanistic understanding of complex electrophysiological events underlying ion transports is still lacking. In addition, non-selective cation (NSC) channels present in the RBC membrane have proven difficult to molecular identification and functional characterization. For instance, NSC channel activity can be elicited by Low Ionic Strength conditions (LIS): the associated change in membrane potential triggers its opening in a voltage dependent manner. But, whereas this depolarizing media produces a spectacular activation of NSC channel, Gárdos channel-evoked hyperpolarization's have been shown to induce sodium entry through a pathway thought to be conductive and termed Pcat. Using the CCCP method, which allows to follow fast changes in membrane potential, we show here (i) that hyperpolarization elicited by Gárdos channel activation triggers sodium entry through a conductive pathway, (ii) that chloride conductance inhibition unveils such conductive cationic conductance, (iii) that the use of the specific chloride conductance inhibitor NS3623 (a derivative of Neurosearch compound NS1652), at concentrations above what is needed for full anion channel block, potentiates the non-selective cation conductance. These results indicate that a non-selective cation channel is likely activated by the changes in the driving force for cations rather than a voltage dependence mechanism per se.

show abstract

Influence of storage and buffer composition on the mechanical behavior of flowing red blood cells

Merlo

Losserand

Yaya

et al. 2023

Biophysical Journal

View full text Add to dashboard Cite

Characterisation of Asp669Tyr Piezo1 cation channel activity in red blood cells: an unexpected phenotype

Cited by 7 publications

References 15 publications

Hereditary Xerocytosis: Differential Behavior of PIEZO1 Mutations in the N-Terminal Extracellular Domain Between Red Blood Cells and HEK Cells

Hereditary Xerocytosis: Differential Behavior of PIEZO1 Mutations in the N-Terminal Extracellular Domain Between Red Blood Cells and HEK Cells

The Chloride Conductance Inhibitor NS3623 Enhances the Activity of a Non-selective Cation Channel in Hyperpolarizing Conditions

Influence of storage and buffer composition on the mechanical behavior of flowing red blood cells

Contact Info

Product

Resources

About