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Characterization of a stretch‐activated potassium channel in chondrocytes
Author(s) -
Mobasheri Ali,
Lewis Rebecca,
Maxwell Judith E.J.,
Hill Claire,
Womack Matthew,
BarrettJolley Richard
Publication year - 2010
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.22075
Subject(s) - bk channel , calcium activated potassium channel , chemistry , potassium channel , patch clamp , biophysics , iberiotoxin , potassium , membrane potential , conductance , ion channel , chondrocyte , sk channel , electrophysiology , extracellular , pipette , anatomy , biochemistry , in vitro , medicine , biology , receptor , mathematics , organic chemistry , combinatorics
Chondrocytes possess the capacity to transduce load‐induced mechanical stimuli into electrochemical signals. The aim of this study was to functionally characterize an ion channel activated in response to membrane stretch in isolated primary equine chondrocytes. We used patch‐clamp electrophysiology to functionally characterize this channel and immunohistochemistry to examine its distribution in articular cartilage. In cell‐attached patch experiments, the application of negative pressures to the patch pipette (in the range of 20–200 mmHg) activated ion channel currents in six of seven patches. The mean activated current was 45.9 ± 1.1 pA ( n = 4) at a membrane potential of 33 mV (cell surface area approximately 240 µm 2 ). The mean slope conductance of the principal single channels resolved within the total stretch‐activated current was 118 ± 19 pS ( n = 6), and reversed near the theoretical potassium equilibrium potential, E K+ , suggesting it was a high‐conductance potassium channel. Activation of these high‐conductance potassium channels was inhibited by extracellular TEA (K d approx. 900 µM) and iberiotoxin (K d approx. 40 nM). This suggests that the current was largely carried by BK‐like potassium (MaxiK) channels. To further characterize these BK‐like channels, we used inside‐out patches of chondrocyte membrane: we found these channels to be activated by elevation in bath calcium concentration. Immunohistochemical staining of equine cartilage samples with polyclonal antibodies to the α1‐ and β1‐subunits of the BK channel revealed positive immunoreactivity for both subunits in superficial zone chondrocytes. These experiments support the hypothesis that functional BK channels are present in chondrocytes and may be involved in mechanotransduction and chemotransduction. J. Cell. Physiol. 223: 511–518, 2010. © 2010 Wiley‐Liss, Inc.