Comparison of M-line and other myofibril components during reversible phorbol ester treatment.

The events occurring during phorbol ester mediated destruction of myofibrils in differentiated muscle cells were followed at the fluorescence and electron microscope levels using antibodies which bind troponin-T, a newly discovered 185 000 dalton M-line protein called myomesin and muscle type creatine kinase. The following series of events is proposed. Within one day of phorbol ester treatment, Z-bands and thin filaments, including troponin-T, are absent from many myofibrils resulting in the rapid loss of longitudinal and lateral alignment. A-bands become randomly oriented and clustered into ever smaller compartments within the rounding, myosac-like, multinucleated cells until after 3 days of treatment they too disappear. The M-line proteins are always present in existing A-bands. These results suggest that the Z-band and associated structures are responsible for the maintenance of alignment and the lateral register of myofibrils, whereas the M-line is responsible for the structural integrity of the A-band. When phorbol ester is removed, the cells revert to a myotube morphology and within 2 to 3 days are filled with myofibrils. A comparison of the appearance of troponin-T and the 185 000 dalton myomesin in the recovery period to their appearance during normal myofibrillogenesis reveals that these proteins are more temporally co-ordinated during myofibrillogenesis than in the phorbol ester experimental system.