Permeabilisation of the sarcolemma in mouse diaphragm exposed to Bay K 8644 in vitro: time course, dependence on Ca2+ and effects of enzyme inhibitors

Treatment of partially depolarised mouse diaphragm muscle in vitro with the Ca2(+)-channel agonist Bay K 8644 (1 microM) induces permeabilisation of the sarcolemma (visualised by penetration of procion yellow). Procion yellow staining was widespread (74% of fibres) after 2 h of treatment, but was negligible after 60 min, a time at which myofibre breakdown is well advanced and elevation of [Ca2+]i is minimal (Howl and Publicover 1989). Permeabilisation was inhibited in Ca2(+)-free saline, and was much less pronounced in polarised fibres. Inhibitors of free radical generation (particularly OH) afforded considerable protection to the muscle membrane against Bay K 8644-induced membrane permeabilisation. Inhibition of phospholipase A2 and lipoxygenase were also effective, but inhibition of xanthine oxidase (by allopurinol) had little effect. It is concluded that the initial effect of Bay K 8644 treatment is to increase Ca2+ influx through Ca2+ channels at the sarcolemma, and that this action subsequently induces membrane permeabilisation. Membrane damage probably occurs due to free radical generation and activation of phospholipase A2, both resulting from elevation of [Ca2+]i.