Ion-dependence of Z-line and M-line response to calcium in striated muscle fibres in rigor.
The calcium-dependent contraction of vertebrate skeletal muscle is thought to be primarily controlled through the interaction of the thick and thin filaments. Through measurement of the Donnan potential, we have shown that an electrical switching mechanism (sensitive to both anions and cations) is present in both A- and I-bands [1]. Here we show that this mechanism is not confined to the contractile apparatus and report for the first time the presence of M-line potentials. The Z-line responds to Ca2+ ions in a similar manner to the A-band under the same solution conditions (phosphate-chloride and imidazole buffers), even though it has no reported Ca2+ binding sites. Z-line potentials were not observed in tris-acetate buffer. The M-line has a markedly different response to any of the other subsarcomeric regions, however, and can only be detected in the phosphate-chloride buffer. Preliminary observations of the M-line potential in creatine kinase-deficient mouse muscle (phosphate-chloride buffer) reveal significant differences in the calcium-induced transitions between two of the genotypes and demonstrate definitively that it is the M-line potential that is being recorded. From these results, it seems likely that the charge response of the Z-line and M-line is being mediated by titin in an anion-dependent manner. Our evidence comes from several observations. First, the similarity between the response of the Z-line potentials to the A-band potentials, where titin is the only link between these structures and second, the differential observation of M-line and Z-line potentials in a range of buffers containing different anion(s). Both Z-line and M-line potentials were seen in phosphate-chloride buffer, but only the Z-line potentials could be detected in chloride-only (imidazole) buffer and neither was observed in the acetate buffer. The latter observations can be attributed to two sources. The first is the effect of acetate buffer on the conformation of myosin [2]; the second is the absence of binding of the M-line protein, myomesin, to titin in the absence of phosphate ions [3].
Coomber
S J
SJ
The Open University Oxford Research Unit, Oxford, UK. s.j.comber@open.ac.uk
Taracewicz
E
E
Akhtar
S
S
deHaan
A
A
Elliott
G F
GF
D000085:Acetates / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002712:Chlorides / Q000494:pharmacology; D064211:Connectin; D007093:Imidazoles / Q000494:pharmacology; D007633:Keratins / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000502:physiology* / Q000648:ultrastructure; D009124:Muscle Proteins / Q000378:metabolism; D018482:Muscle, Skeletal / Q000502:physiology* / Q000648:ultrastructure; D011494:Protein Kinases / Q000378:metabolism; D016658:Psoas Muscles / Q000502:physiology / Q000648:ultrastructure; D011817:Rabbits
D000085:Acetates; D002712:Chlorides; D064211:Connectin; D007093:Imidazoles; D009124:Muscle Proteins; D007633:Keratins; C029899:imidazole; D011494:Protein Kinases; D008274:Magnesium; D002118:Calcium