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Home / Equine Research Bank / Arthritis Rheum / Calcium signaling leads to mitochondrial depolarization in i...
Arthritis and rheumatism2007; 56(7); 2322-2334; doi: 10.1002/art.22717

Calcium signaling leads to mitochondrial depolarization in impact-induced chondrocyte death in equine articular cartilage explants.

Abstract: Chondrocyte apoptosis is an important factor in the progression of osteoarthritis. This study aimed to elucidate the mechanisms involved upstream of caspase 9 activation and, in particular, calcium signaling and mitochondrial depolarization. Methods: Articular cartilage explants obtained from healthy horses were subjected to a single impact load (500-gm weight dropped from a height of 50 mm) and cultured in vitro for up to 48 hours. Chondrocyte death was quantified by the TUNEL method. Release of proteoglycans was determined by the dimethylmethylene blue assay. Weight change was measured, and mitochondrial depolarization was determined using JC-1 staining. To assess the role of calcium signaling in impact-induced chondrocyte death, explants were preincubated in culture medium containing various concentrations of calcium. Inhibitors were used to assess the role of individual signaling components in impact-induced chondrocyte death. Results: Calcium quenching, inhibitors of calpains, calcium/calmodulin-regulated kinase II (CaMKII), and mitochondrial depolarization reduced impact-induced chondrocyte death after 48 hours in culture. Transient mitochondrial depolarization was observed 3-6 hours following a single impact load. Mitochondrial depolarization was prevented by calcium quenching, inhibitors of calpain, CaMKII, permeability transition pore formation, ryanodine receptor, and the mitochondrial uniport transporter. Cathepsin B did not appear to be involved in impact-induced chondrocyte death. The calpain inhibitor prevented proteoglycan loss, but the percentage weight gain and proteoglycan loss were unaffected by all treatments used. Conclusions: Following a single impact load, calcium is released from the endoplasmic reticulum via the ryanodine receptor and is taken up by the mitochondria via the uniport transporter, causing mitochondrial depolarization and caspase 9 activation. In addition, calpains and CaMKII play important roles in causing mitochondrial depolarization.
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Publication Date: 2007-06-30 PubMed ID: 17599752DOI: 10.1002/art.22717Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 research study explores how calcium signaling leads to cell death in cartilage tissues following a physical impact. Through their experiments, they demonstrate various mechanisms involved in this process and their relationship to osteoarthritis.

Research Context

  • The study aimed to understand the mechanisms leading to the death of chondrocytes (cartilage cells), particularly focusing on calcium signaling and mitochondrial depolarization.
  • The research has a key relevance to osteoarthritis, a common joint condition, as chondrocyte apoptosis (cell death) significantly contributes to its progression.

Methodology

  • Horse articular cartilage explants were subjected to an impact load and then cultured in vitro for up to 48 hours.
  • The rate of chondrocyte death was quantified using the TUNEL method, and the release of proteoglycans was measured using a specific assay.
  • The researchers manipulated the calcium concentration in the culture medium and used inhibitors targeting specific components of the signaling pathway to understand their roles in chondrocyte death.

Results

  • The study found that calcium quenching as well as inhibiting certain cellular components could reduce impact-induced chondrocyte death. These included calpains (calcium activated enzymes), CaMKII (a calcium/calmodulin regulated enzyme), and elements associated with mitochondrial depolarization.
  • The temporary mitochondrial depolarization observed 3-6 hours after impact was prevented by the same treatments.
  • The results suggest that calcium is released from the endoplasmic reticulum via a specific receptor following impact and taken up by the mitochondria, causing mitochondrial depolarization and triggering the activation of caspase 9 – a key enzyme in the apoptosis process.
  • The enzyme cathepsin B, contrary to some expectations, did not appear to play a role in chondrocyte death.

Conclusions

  • The study concludes that following a physical impact, calcium signaling can lead to mitochondrial depolarization and caspase 9 activation, causing chondrocyte death.
  • This process appears to be heavily influenced by the enzymes calpains and CaMKII.
  • These findings provide new insights for the development of potential treatments to minimize cell death following joint injuries and slow the progression of conditions such as osteoarthritis.

Cite This Article

APA
Huser CA, Davies ME. (2007). Calcium signaling leads to mitochondrial depolarization in impact-induced chondrocyte death in equine articular cartilage explants. Arthritis Rheum, 56(7), 2322-2334. https://doi.org/10.1002/art.22717

Publication

Arthritis and rheumatism
ISSN: 0004-3591
NlmUniqueID: 0370605
Country: United States
Language: English
Volume: 56
Issue: 7
Pages: 2322-2334

Researcher Affiliations

Huser, C A M
  • University of Cambridge, Cambridge, UK. C.Huser@clinmed.gla.ac.uk
Davies, M E

    MeSH Terms

    • Animals
    • Calcium / metabolism
    • Calcium Signaling / drug effects
    • Calcium Signaling / physiology
    • Cartilage, Articular / drug effects
    • Cartilage, Articular / physiology
    • Cathepsin B / antagonists & inhibitors
    • Dipeptides / pharmacology
    • Horses
    • Mitochondria / physiology
    • Organ Culture Techniques
    • Reference Values
    • Weight Gain
    • Weight-Bearing

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