Hypoosmotic cell swelling as a novel mechanism for modulation of cloned HCN2 channels.
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Hypoosmotic cell swelling as a novel mechanism for modulation of cloned HCN2 channels. / Calloe, Kirstine; Elmedyb, Pernille; Olesen, Søren-Peter; Jorgensen, Nanna K; Grunnet, Morten.
In: Biophysical Journal, Vol. 89, No. 3, 2005, p. 2159-69.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Hypoosmotic cell swelling as a novel mechanism for modulation of cloned HCN2 channels.
AU - Calloe, Kirstine
AU - Elmedyb, Pernille
AU - Olesen, Søren-Peter
AU - Jorgensen, Nanna K
AU - Grunnet, Morten
N1 - Keywords: Actins; Animals; Cell Shape; Cyclic AMP; Cytochalasin D; Cytoskeleton; DNA, Complementary; Electrophysiology; Globins; Humans; Ion Channels; Ions; KCNQ Potassium Channels; Kinetics; Oocytes; Osmosis; Phalloidine; Time Factors; Xenopus laevis
PY - 2005
Y1 - 2005
N2 - This work demonstrates cell swelling as a new regulatory mechanism for the cloned hyperpolarization-activated, cyclic nucleotide-gated channel 2 (HCN2). HCN2 channels were coexpressed with aquaporin1 in Xenopus laevis oocytes and currents were monitored using a two-electrode voltage-clamp. HCN2 channels were activated by hyperpolarization to -100 mV and the currents were measured before and during hypoosmotic cell swelling. Cell swelling increased HCN2 currents by 30% without changing the kinetics of the currents. Injection of 50 nl intracellular solution resulted in a current increase of 20%, indicating that an increase in cell volume also under isoosmotic conditions may lead to activation of HCN2. In the absence of aquaporin1 only negligible changes in oocyte cell volume occur during exposure to hypoosmotic media and no significant change in HCN2 channel activity was observed during perfusion with hypoosmotic media. This indicates that cell swelling and not a change in ionic strength of the media, caused the observed swelling-induced increase in current. The increase in HCN2 current induced by cell swelling could be abolished by cytochalasin D treatment, indicating that an intact F-actin cytoskeleton is a prerequisite for the swelling-induced current.
AB - This work demonstrates cell swelling as a new regulatory mechanism for the cloned hyperpolarization-activated, cyclic nucleotide-gated channel 2 (HCN2). HCN2 channels were coexpressed with aquaporin1 in Xenopus laevis oocytes and currents were monitored using a two-electrode voltage-clamp. HCN2 channels were activated by hyperpolarization to -100 mV and the currents were measured before and during hypoosmotic cell swelling. Cell swelling increased HCN2 currents by 30% without changing the kinetics of the currents. Injection of 50 nl intracellular solution resulted in a current increase of 20%, indicating that an increase in cell volume also under isoosmotic conditions may lead to activation of HCN2. In the absence of aquaporin1 only negligible changes in oocyte cell volume occur during exposure to hypoosmotic media and no significant change in HCN2 channel activity was observed during perfusion with hypoosmotic media. This indicates that cell swelling and not a change in ionic strength of the media, caused the observed swelling-induced increase in current. The increase in HCN2 current induced by cell swelling could be abolished by cytochalasin D treatment, indicating that an intact F-actin cytoskeleton is a prerequisite for the swelling-induced current.
U2 - 10.1529/biophysj.105.063792
DO - 10.1529/biophysj.105.063792
M3 - Journal article
C2 - 15980171
VL - 89
SP - 2159
EP - 2169
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 3
ER -
ID: 8418798