Muscle-specific calpain is localized in regions near motor endplates in differentiating lobster claw muscles.
Calpains are Ca2+-dependent proteinases that mediate protein turnover in crustacean skeletal muscles. We used an antibody directed against lobster muscle-specific calpain (Ha-CalpM) to examine its distribution in differentiating juvenile lobster claw muscles. These muscles are comprised of both fast and slow fibers early in development, but become specialized into predominantly fast or exclusively slow muscles in adults. The transition into adult muscle types requires that myofibrillar proteins specific for fast or slow muscles to be selectively removed and replaced by the appropriate proteins. Using immunohistochemistry, we observed a distinct staining pattern where staining was preferentially localized in the fiber periphery along one side of the fiber. Immunolabeling with an antibody directed against synaptotagmin revealed that the calpain staining was greatest in the cytoplasm adjacent to synaptic terminals. In complementary analyses, we used sequence-specific primers with real-time PCR to quantify the levels of Ha-CalpM in whole juvenile claw muscles. These expression levels were not significantly different between cutter and crusher claws, but were positively correlated with the expression of fast myosin heavy chain. The anatomical localization of Ha-CalpM near motor endplates, coupled with the correlation with fast myofibrillar gene expression, suggests a role for this intracellular proteinase in fiber type switching.
Medler
Scott
S
Department of Biology, Colorado State University, Fort Collins, Colorado 80523, USA. smedler@buffalo.edu
Chang
Ernest S
ES
Mykles
Donald L
DL
D000375:Aging / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002154:Calpain / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D006724:Hoof and Claw / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D007150:Immunohistochemistry; D009045:Motor Endplate / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D024510:Muscle Development / Q000235:genetics; D018482:Muscle, Skeletal / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000378:metabolism; D008121:Nephropidae / Q000166:cytology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050857:Synaptotagmins / Q000378:metabolism