Intracellular pH gradients in migrating cells
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Intracellular pH gradients in migrating cells. / Martin, Christine; Pedersen, Stine Helene Falsig; Schwab, Albrecht; Stock, Christian.
In: American Journal of Physiology: Cell Physiology, Vol. 300, No. 3, 2011, p. C490-C495.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Intracellular pH gradients in migrating cells
AU - Martin, Christine
AU - Pedersen, Stine Helene Falsig
AU - Schwab, Albrecht
AU - Stock, Christian
PY - 2011
Y1 - 2011
N2 - Cell polarization along the axis of movement is required for migration. The localization of proteins and regulators of the migratory machinery to either the cell front or its rear results in a spatial asymmetry enabling cells to simultaneously coordinate cell protrusion and retraction. Protons might function as such unevenly distributed regulators as they modulate the interaction of focal adhesion proteins and components of the cytoskeleton in vitro. However, an intracellular pH (pH(i)) gradient reflecting a spatial asymmetry of protons has not been shown so far. One major regulator of pH(i), the Na(+)/H(+) exchanger NHE1, is essential for cell migration and accumulates at the cell front. Here, we test the hypothesis that the uneven distribution of NHE1 activity creates a pH(i) gradient in migrating cells. Using the pH-sensitive fluorescent dye BCECF, pH(i) was measured in five cell lines (MV3, B16V, NIH3T3, MDCK-F1, EA.hy926) along the axis of movement. Differences in pH(i) between the front and the rear end (¿pH(i) front-rear) were present in all cell lines, and inhibition of NHE1 either with HOE642 or by absence of extracellular Na(+) caused the pH(i) gradient to flatten or disappear. In conclusion, pH(i) gradients established by NHE1 activity exist along the axis of movement.
AB - Cell polarization along the axis of movement is required for migration. The localization of proteins and regulators of the migratory machinery to either the cell front or its rear results in a spatial asymmetry enabling cells to simultaneously coordinate cell protrusion and retraction. Protons might function as such unevenly distributed regulators as they modulate the interaction of focal adhesion proteins and components of the cytoskeleton in vitro. However, an intracellular pH (pH(i)) gradient reflecting a spatial asymmetry of protons has not been shown so far. One major regulator of pH(i), the Na(+)/H(+) exchanger NHE1, is essential for cell migration and accumulates at the cell front. Here, we test the hypothesis that the uneven distribution of NHE1 activity creates a pH(i) gradient in migrating cells. Using the pH-sensitive fluorescent dye BCECF, pH(i) was measured in five cell lines (MV3, B16V, NIH3T3, MDCK-F1, EA.hy926) along the axis of movement. Differences in pH(i) between the front and the rear end (¿pH(i) front-rear) were present in all cell lines, and inhibition of NHE1 either with HOE642 or by absence of extracellular Na(+) caused the pH(i) gradient to flatten or disappear. In conclusion, pH(i) gradients established by NHE1 activity exist along the axis of movement.
KW - Animals
KW - Cation Transport Proteins
KW - Cell Line, Transformed
KW - Cell Line, Tumor
KW - Cell Movement
KW - Cell Polarity
KW - Cell Surface Extensions
KW - Dogs
KW - Fluoresceins
KW - Fluorescent Dyes
KW - Humans
KW - Hydrogen-Ion Concentration
KW - Intracellular Fluid
KW - Mice
KW - NIH 3T3 Cells
KW - Protons
KW - Sodium-Hydrogen Antiporter
U2 - 10.1152/ajpcell.00280.2010
DO - 10.1152/ajpcell.00280.2010
M3 - Journal article
C2 - 21148407
VL - 300
SP - C490-C495
JO - American Journal of Physiology: Cell Physiology
JF - American Journal of Physiology: Cell Physiology
SN - 0363-6143
IS - 3
ER -
ID: 33345437