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Elife ; 72018 08 22.
Article in English | MEDLINE | ID: mdl-30132757

ABSTRACT

Piezo1 is a mechanosensitive (MS) ion channel with characteristic fast-inactivation kinetics. We found a slowly-inactivating MS current in mouse embryonic stem (mES) cells and characterized it throughout their differentiation into motor-neurons to investigate its components. MS currents were large and slowly-inactivating in the stem-cell stage, and became smaller and faster-inactivating throughout the differentiation. We found that Piezo1 is expressed in mES cells, and its knockout abolishes MS currents, indicating that the slowly-inactivating current in mES cells is carried by Piezo1. To further investigate its slow inactivation in these cells, we cloned Piezo1 cDNA from mES cells and found that it displays fast-inactivation kinetics in heterologous expression, indicating that sources of modulation other than the aminoacid sequence determine its slow kinetics in mES cells. Finally, we report that Piezo1 knockout ES cells showed a reduced rate of proliferation but no significant differences in other markers of pluripotency and differentiation.


Subject(s)
Ion Channel Gating , Ion Channels/metabolism , Mechanotransduction, Cellular , Mouse Embryonic Stem Cells/metabolism , Animals , Base Sequence , Cell Differentiation , Cell Proliferation , Cell Shape , DNA, Complementary/genetics , Gastrulation , HEK293 Cells , Humans , Kinetics , Mice , Mice, Knockout , Motor Neurons/cytology , Motor Neurons/metabolism , Mouse Embryonic Stem Cells/cytology , Mutation/genetics , Phenotype , Pluripotent Stem Cells/cytology , Pluripotent Stem Cells/metabolism
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