Effects of internal Sr2+ on the activity of large-conductance Ca2+-activated K+ channels were studied in inside-out membrane patches from goldfish saccular hair cells. Sr2+ was approximately one-fourth as potent as Ca2+ in activating these channels. Although the Hill coefficient for Sr2+ was smaller than that for Ca2+, maximum open-state probability, voltage dependence, steady state gating kinetics, and time courses of activation and deactivation of the channel were very similar under the presence of equipotent concentrations of Ca2+ and Sr2+. This suggests that voltage-dependent activation is partially independent of the ligand. Internal Sr2+ at higher concentrations (>100 μM) produced fast and slow blockade both concentration and voltage dependently. The reduction in single-channel amplitude (fast blockade) could be fitted with a modified Woodhull equation that incorporated the Hill coefficient. The dissociation constant at 0 mV, the Hill coefficient, and zd (a product of the charge of the blocking ion and the fraction of the voltage difference at the binding site from the inside) in this equation were 58–209 mM, 0.69–0.75, 0.45–0.51, respectively (n = 4). Long shut events (slow blockade) produced by Sr2+ lasted ∼10–200 ms and could be fitted with single-exponential curves (time constant, τl−s) in shut-time histograms. Durations of burst events, periods intercalated by long shut events, could also be fitted with single exponentials (time constant, τb). A significant decrease in τb and no large changes in τl−s were observed with increased Sr2+ concentration and voltage. These findings on slow blockade could be approximated by a model in which single Sr2+ ions bind to a blocking site within the channel pore beyond the energy barrier from the inside, as proposed for Ba2+ blockade. The dissociation constant at 0 mV and zd in the Woodhull equation for this model were 36–150 mM and 1–1.8, respectively (n = 3).
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1 February 1998
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February 01 1998
Activation and Two Modes of Blockade by Strontium of Ca2+-activated K+ Channels in Goldfish Saccular Hair Cells
Izumi Sugihara
Izumi Sugihara
From the Department of Physiology, Tokyo Medical and Dental University School of Medicine, Tokyo 113, Japan
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Izumi Sugihara
From the Department of Physiology, Tokyo Medical and Dental University School of Medicine, Tokyo 113, Japan
Address correspondence to Dr. Izumi Sugihara, Department of Physiology, Tokyo Medical and Dental University, School of Medicine, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113, Japan. Fax: 81-3-5803-5155; E-mail: [email protected]
The author thanks Dr. Taro Furukawa for his advice and encouragement regarding this work.
Received:
June 30 1997
Accepted:
October 21 1997
Online ISSN: 1540-7748
Print ISSN: 0022-1295
1998
J Gen Physiol (1998) 111 (2): 363–379.
Article history
Received:
June 30 1997
Accepted:
October 21 1997
Citation
Izumi Sugihara; Activation and Two Modes of Blockade by Strontium of Ca2+-activated K+ Channels in Goldfish Saccular Hair Cells . J Gen Physiol 1 February 1998; 111 (2): 363–379. doi: https://doi.org/10.1085/jgp.111.2.363
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