Eyestalk ablation has little effect on actin and myosin heavy chain gene expression in adult lobster skeletal muscles.
The organization of skeletal muscles in decapod crustaceans is significantly altered during molting and development. Prior to molting, the claw muscles atrophy dramatically, facilitating their removal from the base of the claw. During development, lobster claw muscles exhibit fiber switching over several molt cycles. Such processes may be influenced by the secretion of steroid molting hormones, known collectively as ecdysteroids. To assay the effects of these hormones, we used eyestalk ablation to trigger an elevation of circulating ecdysteroids and then quantified myofibrillar mRNA levels with real-time PCR and myofibrillar protein levels by SDS-PAGE. Levels of myosin heavy chain (MHC) and actin proteins and the mRNA encoding them were largely unaffected by eyestalk ablation, but in muscles from intact animals, myofibrillar gene expression was modestly elevated in premolt and postmolt animals. In contrast, polyubiquitin mRNA was significantly elevated (about 2-fold) in claw muscles from eyestalk-ablated animals with elevated circulating ecdysteroids. Moreover, patterns of MHC and actin gene expression are significantly different among slow and fast claw muscles. Consistent with these patterns, the three muscle types differed in the relative amounts of myosin heavy chain and actin proteins. All three muscles also co-expressed fast and slow myosin isoforms, even in fibers that are generally regarded as exclusively fast or slow. These results are consistent with other recent data demonstrating co-expression of myosin isoforms in lobster muscles.
Medler
Scott
S
Department of Biology, Colorado State University, Fort Collins, Colorado 80523, USA. smedler@buffalo.edu
Brown
Kitty J
KJ
Chang
Ernest S
ES
Mykles
Donald L
DL
D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D002140:California; D026461:Ecdysteroids / Q000097:blood / Q000378:metabolism* / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D007447:Invertebrate Hormones / Q000172:deficiency* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism*; D008121:Nephropidae / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins; D026461:Ecdysteroids; D007447:Invertebrate Hormones; C012445:molt-inhibiting hormone; D018995:Myosin Heavy Chains