1 We examined the effect of the protein kinase C (PKC) inhibitor chelerythrine on depolarization-induced force responses (DIFRs) and sarcoplasmic reticulum (SR) function in single, mechanically skinned skeletal muscle fibres of the rat.2 In this study, the DIFRs in the skinned fibres normally underwent an irreversible loss of excitation-contraction coupling (ECC) after 10-15 responses. Chelerythrine (12 muM) was shown to restore ECC in these fibres. Restored force responses were similar in peak (control 50.8+/-6.4%, chelerythrine 56.9+/-12.4% of maximum force, P=0.42, n=21), but significantly broadened compared to initial control responses (full-width at half maximum, control; 3.7+/-0.3 s, chelerythrine; 13.3+/-1.1 s, P<0.001). Early exposure to chelerythrine prevented run-down of DIFRs. Chelerythrine also induced spontaneous force responses in some fibres.3 The PKC inhibitors calphostin C and staurosporine did not restore ECC, and the PKC activator phorbol 12-myristate 13-acetate did not promote loss of ECC in the skinned fibres.4 Chelerythrine significantly increased SR Ca2+ loading by 8.4+/-1.7% (P=0.02, n=9) and SR Ca2+ release by at least 14.1+/-2.7% (P = 0.004, n 11) in the skinned fibres.5 Chelerythrine had no significant effect on maximum force production or the [Ca2+] producing half maximal activation of the myofilaments. However, chelerythrine did have a small effect on the slope of the force-Ca2+ relationship (P=0.02, n=10).6 Chelerythrine reverses the use-dependent loss of excitation-contraction coupling in skinend skeletal muscle fibres by a PKC independent pathway. Chelerythrine may be an important pharmacological probe for examining the mechanisms of contraction-induced muscle injury.