Myosin heavy chain isoform expression and Ca2+-stimulated ATPase activity in single fibres of toad rectus abdominis muscle

Segments of single fibres from the rectus abdominis (RA) muscles of adult and juvenile cane toads (Bufo marinus) were examined for myosin heavy chain (mHC) isoform expression and Ca2+-stimulated MgATPase activity. mHC isoform analyses were carried out using the recently developed alanine-SDS-PAGE method, which separates one tonic (BmHCT) and three twitch (BmHC1, BmHC2, BmHC3) mHC isoforms in toad skeletal muscle. Ca2+-stimulated MgATPase activity was measured by spectrophotometric determination of P-i, under conditions in which the ATPase associated with the sarcoplasmic reticulum (SR ATPase) was suppressed by feedback inhibition. The mHC-based fibre types identified in this study include three pure twitch fibre types (t1, t2 and t3), expressing BmHC1, BmHC2 or BmHC3 respectively, and seven hybrid fibre types co-expressing a combination of two or three twitch and tonic or twitch and twitch mHC isoforms. The fibre populations dissected from juvenile and adult toad muscles contained 49.4% (juvenile) and 73.7% (adult) mHC hybrids. The average values for Ca2+-stimulated MgATPase in pure twitch fibres and in fibres expressing predominantly (greater than or equal to95%) the tonic mHC isoform (T-p fibres) differed significantly (P < 0.05) from each other and decreased in the order t1 > t2 > t3 > T-p. We conclude that (i) in RA muscles of both juvenile and adult cane toads there is a large proportion of mHC hybrids, some of which co-express twitch and tonic mHC isoforms and (ii) ATPase activities associated with the four mHC isoforms expressed in toad skeletal muscles decrease in the order BmHC1 > BmHC2 > BmHC3 > BmHCT.