Real time RT-PCR with a newly designed set of primers confirmed the presence of 2b and 2x/d myosin heavy chain mRNAs in the rat slow soleus muscle.
In order to re-evaluate the presence and relative quantity of 2b and 2x/d myosin heavy chain (MyHC) transcripts in rat slow soleus muscle by using real time RT-PCR we have compared the available relevant cDNA sequences and designed a new set of primers having similar melting temperatures, matching separate MyHC exons in the regions of maximal differences in MyHC coding sequences, and containing G or C at the 3 -end. These also yielded PCR products of corresponding length, which is an important requirement for real time RT-PCR quantification. The experiments were performed on 8-month-old inbred female Lewis strain rats used in our current study of regenerating transplanted muscles. The real time RT-PCR measurement confirmed the expression of all four MyHC mRNAs (type 1, 2a, 2x/d and 2b) in both fast extensor digitorum longus and slow soleus muscles, although in the soleus muscle of adult rats, only type 1 and 2a protein isoforms can be usually detected.
Žurmanová
J
J
Institute of Physiology, Czech Academy of Sciences, Prague, Czech Republic.
Půta
F
F
Stopková
R
R
Soukup
T
T
D000818:Animals; D017931:DNA Primers; D005260:Female; D018656:Muscle Fibers, Fast-Twitch / Q000737:chemistry; D018657:Muscle Fibers, Slow-Twitch / Q000737:chemistry*; D018482:Muscle, Skeletal / Q000737:chemistry*; D018995:Myosin Heavy Chains / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D011917:Rats, Inbred Lew
D017931:DNA Primers; D020033:Protein Isoforms; D012333:RNA, Messenger; D018995:Myosin Heavy Chains