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Related Experiment Videos

Morphological changes in cultured myotubes treated with agents that interfere with lysosomal function

S Bursztajn, P Libby

    Cell and Tissue Research
    |January 1, 1981
    PubMed
    Summary

    Lysosomal inhibitors like chloroquine and leupeptin prevent acetylcholine receptor (AChR) degradation in muscle cells. This leads to receptor buildup and increased coated vesicles, suggesting impaired lysosome-vesicle fusion.

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    Area of Science:

    • Cell biology
    • Muscle cell physiology
    • Receptor trafficking

    Background:

    • Lysosomes degrade cellular components, including acetylcholine receptors (AChR).
    • Inhibitors of lysosomal function impede this degradation process.
    • Coated vesicles are implicated in intracellular transport.

    Purpose of the Study:

    • To investigate the effects of leupeptin and chloroquine on cultured myotube morphology.
    • To understand acetylcholine receptor turnover and the origin of coated vesicles.
    • To elucidate the mechanism by which lysosomal inhibitors affect receptor degradation.

    Main Methods:

    • Treatment of cultured myotubes with leupeptin and chloroquine.
    • Morphological analysis using microscopy.

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  • Quantification of coated vesicle size and content (horseradish peroxidase uptake).

    • Main Results:

    • Chloroquine induced plasma membrane involution, sarcomere disorganization, vacuolization, and enlarged lysosome-like bodies.
    • Leupeptin enlarged dense bodies but did not cause vacuolization or sarcomere disruption.
    • Chloroquine treatment led to accumulation of large coated vesicles, while leupeptin treatment resulted in smaller coated vesicles. Chloroquine-treated cells showed increased horseradish peroxidase uptake in coated vesicles.

    Conclusions:

    • Chloroquine interferes with AChR degradation by inhibiting lysosome-coated vesicle fusion.
    • Leupeptin and chloroquine differentially affect myotube morphology and coated vesicle characteristics.
    • These findings provide insights into receptor turnover and the role of coated vesicles in muscle cells.