Existence of voltage-dependent Ca2+ channels in vestibular dark cells: cytochemical and whole-cell patch-clamp studies

1997 ◽  
Vol 254 (6) ◽  
pp. 287-291 ◽  
Author(s):  
K. Imon ◽  
T. Amano ◽  
K. Ishihara ◽  
M. Sasa ◽  
K. Yajin
Keyword(s):  
Patch Clamp ◽  
Whole Cell ◽  
Dark Cells ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Voltage Dependent ◽  
Vestibular Dark Cells ◽  
Ca2 Channels

scholarly journals Kvβ1.3 Reduces the Degree of Stereoselective Bupivacaine Block of Kv1.5 Channels

2007 ◽  
Vol 107 (4) ◽  
pp. 641-651 ◽  
Author(s):  
Cristina Arias ◽  
Miriam Guizy ◽  
Miren David ◽  
Stefanie Marzian ◽  
Teresa González ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Binding Site ◽  
Voltage Clamp ◽  
Xenopus Oocytes ◽  
Time Dependent ◽  
Hek293 Cells ◽  
Whole Cell ◽  
Cell Patch Clamp ◽  
Whole Cell Patch Clamp ◽  
Voltage Dependent

Background Kvbeta1.3 subunit modifies the gating and the pharmacology of Kv1.5 channels, decreasing their sensitivity to block induced by drugs, suggesting that Kvbeta1.3 competes with them for a binding site at Kv1.5 channels. Methods Currents generated by the activation of Kv1.5 and Kv1.5 + Kvbeta1.3 channels expressed in HEK293 cells and Xenopus oocytes were recorded by using whole cell patch clamp and voltage clamp techniques. Results Block of Kv1.5, but not that produced on Kv1.5 + Kvbeta1.3 channels, was voltage dependent. In both channels, bupivacaine block was time dependent. R(+)- and S(-)-bupivacaine blocked Kv1.5 with IC50 4.4 +/- 0.5 microM (n = 15) and 39.8 +/- 8.2 microM (n = 16; P < 0.05), respectively. These values increased fourfold for R(+)-bupivacaine (17.2 +/- 2.2 microM) and twofold for S(-)-bupivacaine (71.9 +/- 11.5 microM) in Kv1.5 + Kvbeta1.3 channels. Therefore, the degree of stereoselectivity (theta) decreased from 9 to 4 in the presence of Kvbeta1.3. The decrease in potency to block Kv1.5 + Kvbeta1.3 channels was the result of a less stable interaction between bupivacaine enantiomers and channels. Differences in stereoselectivity in each situation were due to a more favorable interaction between the channel and R(+)-bupivacaine. In the presence of Kvbeta1.3, stereoselectivity was abolished for V514A mutant channels (involved in bupivacaine binding but not in Kvbeta1.3 binding) but not for L510A (part of Kvbeta1.3 binding site). Conclusions The degree of stereoselective block of Kv1.5 decreases from 9 to 4 when Kvbeta1.3 is present. L510 is determinant for the modulation of bupivacaine block, because it is the only residue of the S6 segment that binds to both bupivacaine and Kvbeta1.3. These findings support an overlapping binding site for drugs and Kvbeta1.3.

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