SUMMARY Although muscle specialization has been studied extensively in vertebrates,less is known about the mechanisms that have evolved in invertebrate muscle that modulate muscle performance. Recent research on the musculature of squid suggests that the mechanisms of muscle specialization in cephalopods may differ from those documented in vertebrates. Muscle diversity in the development and the evolution of cephalopods appears to be characterized by modulation of the dimensions of the myofilaments, in contrast to the relatively fixed myofilament dimensions of vertebrate muscle. In addition, the arrangement of the myofilaments may also be altered, as has been observed in the extensor muscle fibres of the prey capture tentacles of squid and cuttlefish, which show cross-striation and thus differ from the obliquely striated pattern of most cephalopod locomotor muscle fibres. Although biochemical specializations that reflect differences in aerobic capacity have been documented previously for specific layers of the mantle muscle of squid,comparison of protein profiles of myofilament preparations from the fast cross-striated tentacle fibres and slow obliquely striated fibres from the arms has revealed remarkably few differences in myofilament lattice proteins. In particular, previous studies using a variety of SDS-PAGE techniques and peptide mapping of the myosin heavy chain were unable to resolve differences in the myosin light and heavy chains. Since these techniques cannot exclude the presence of a highly conserved variant that differs in only a few amino acids, in this study semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis of myosin heavy chain messenger RNAs (mRNAs) from the cross-striated tentacle and obliquely striated arm muscle fibres was conducted. This analysis showed that a previously reported alternatively spliced isoform of the squid myosin motor domain is present only in low abundance in both muscle types and therefore differential expression of the two myosins cannot explain the difference in contractile properties. It thus appears that modulation of the contractile properties of the musculature of squid and other cephalopods occurs primarily through variation in the arrangement and dimensions of the myofilaments.