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Journal of cellular physiology2010; 223(2); 511-518; doi: 10.1002/jcp.22075

Characterization of a stretch-activated potassium channel in chondrocytes.

Abstract: 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 microm(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 microM) 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 alpha1- and beta1-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.
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Publication Date: 2010-02-18 PubMed ID: 20162564PubMed Central: PMC2883078DOI: 10.1002/jcp.22075Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study investigates how certain cells in horses known as chondrocytes can transmit mechanical energy into electrical signals. The researchers examined a specific ion channel in these cells that responds to physical stress and identified it as a high-conductance potassium channel. Their findings suggest these channels could play a role in converting mechanical and chemical stimuli into cellular responses.

Overview of Research

  • The scientists’ objective was to explore the functionality of an ion channel that gets activated when a membrane stretch occurs in isolated, primary equine chondrocytes. Chondrocytes are cells in cartilage tissue that have the ability to change mechanical stimuli (pressure changes) into electrochemical signals (movement of ions across their membranes).
  • The analysis used patch-clamp electrophysiology, a method that measures the electric current in individual ion channels. Also, they used immunohistochemistry, a staining technique, to examine the ion channel’s distribution in the cartilage tissue.

Experiment Findings

  • In six out of seven tests using negative pressure applied to the patch pipette, ion channel currents activated. The average activated current measured 45.9 +/- 1.1 pA at a membrane potential of 33 mV.
  • The channel closely resembled what’s known as a high-conductance potassium channel based on their calculated mean slope conductance and the fact that they reversed near the theoretical potassium equilibrium potential.

Identification of the High-Conductance Potassium Channel

  • Using external TEA (a potassium channel blocker) and iberiotoxin (a selective blocker of a certain type of potassium channel), they inhibited the activation of these high-conductance potassium channels.
  • The researchers found out that the ion channel activity was largely carried by BK-like potassium (MaxiK) channels, which are high-conductance potassium channels sensitive to both voltage and calcium ions.
  • They utilized inside-out patches of chondrocyte membrane and discovered that these BK-like channels were prompted by a rise in bath calcium concentration.

Immunohistochemical Staining

  • Equine cartilage samples were stained with antibodies for both subunits of the BK channel. The resulting positive reaction for both subunits in superficial zone chondrocytes confirmed the presence of functional BK channels.

Conclusion and Implications

  • This study supports the possibility that BK-like potassium (MaxiK) channels are present and functional in chondrocytes, where they could be involved in converting mechanical and chemical stimuli into cellular responses (mechanotransduction and chemotransduction).
  • Understanding these processes can potentially provide insights into cartilage function and pathology-such as osteoarthritis-, and even guide future therapeutic strategies in veterinary and human medicine.

Cite This Article

APA
Mobasheri A, Lewis R, Maxwell JE, Hill C, Womack M, Barrett-Jolley R. (2010). Characterization of a stretch-activated potassium channel in chondrocytes. J Cell Physiol, 223(2), 511-518. https://doi.org/10.1002/jcp.22075

Publication

Journal of cellular physiology
ISSN: 1097-4652
NlmUniqueID: 0050222
Country: United States
Language: English
Volume: 223
Issue: 2
Pages: 511-518

Researcher Affiliations

Mobasheri, Ali
  • Musculoskeletal Research Group, Division of Veterinary Medicine, Faculty of Medicine and Health Sciences, University of Nottingham, Leicestershire, United Kingdom.
Lewis, Rebecca
    Maxwell, Judith E J
      Hill, Claire
        Womack, Matthew
          Barrett-Jolley, Richard

            MeSH Terms

            • Animals
            • Cartilage / cytology
            • Cartilage / metabolism
            • Cell Membrane / drug effects
            • Cell Membrane / metabolism
            • Cell Membrane / ultrastructure
            • Chondrocytes / cytology
            • Chondrocytes / drug effects
            • Chondrocytes / metabolism
            • Horses
            • Ion Channel Gating / drug effects
            • Ion Channel Gating / physiology
            • Large-Conductance Calcium-Activated Potassium Channels / drug effects
            • Large-Conductance Calcium-Activated Potassium Channels / metabolism
            • Mechanotransduction, Cellular / physiology
            • Membrane Potentials / drug effects
            • Membrane Potentials / physiology
            • Organ Culture Techniques
            • Patch-Clamp Techniques
            • Potassium / metabolism
            • Potassium Channel Blockers / pharmacology
            • Pressure / adverse effects
            • Protein Subunits / drug effects
            • Protein Subunits / metabolism
            • Stress, Mechanical
            • Weight-Bearing / physiology

            Grant Funding

            • Biotechnology and Biological Sciences Research Council
            • Wellcome Trust

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