Identification of myosin heavy chain I, IIa and IIx in canine skeletal muscles by an electrophoretic and immunoblotting study.
To determine which myosin heavy chain (MHC) isoforms are expressed in canine skeletal muscles, different muscle samples of five mixed-breed dogs were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The separated MHC isoforms were identified by immunoblotting technique using a set of specific monoclonal antibodies. To compare the results of the electrophoretic and immunoblotting study, the pattern of MHC isoform expression and histochemical profiles of canine fibres were additionally demonstrated on serial muscle sections by immunohistochemistry and myofibrillar adenosine triphosphatase (mATPase) histochemistry. Not more than three MHC isoforms were demonstrated by SDS-PAGE in the analysed canine muscles. By the immunoblotting technique, the fastest migrating MHC band was identified as slow or MHC-I, the intermediate one as MHC-IIx and the slowest migrating band as MHC-IIa isoform. Since none of the three MHC bands and none of the analysed fibres were recognized by the antibody specific to MHC-IIb of rats, we concluded that MHC-IIb is not expressed in large skeletal muscles of dogs. Similarly, only three major fibre types, i.e. I, IIA and IIX, were revealed according to the pattern of MHC immunohistochemistry and mATPase reaction. Type IIA fibres were more alkali- and acid-stable than type IIX fibres after mATPase histochemistry; hence, the latter corresponded to type IIDog fibres. However, beside the three major fibre types, scarce hybrid fibres co-expressing two MHC isoforms (I/IIA and IIA/IIX) were demonstrated by immunohistochemistry.
Smerdu
V
V
Institute of Anatomy, Medical Faculty, University in Ljubljana, Ljubljana, Slovenia. vika.smerdu@mf.uni-lj.si
Strbenc
M
M
Meznaric-Petrusa
M
M
Fazarinc
G
G
D000818:Animals; D000911:Antibodies, Monoclonal; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000145:classification* / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism; D020033:Protein Isoforms / Q000378:metabolism; D051381:Rats
D000911:Antibodies, Monoclonal; D020033:Protein Isoforms; D018995:Myosin Heavy Chains; D009218:Myosins