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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
J. Žurmanová, F. Půta, R. Stopková, T. Soukup
Language English Country Czech Republic
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- Keywords
- Rat slow soleus muscle, Rat fast EDL muscle, Myosin heavy chain isoforms, Real time RT-PCR,
- MeSH
- DNA Primers MeSH
- Financing, Organized MeSH
- Muscle, Skeletal chemistry MeSH
- Rats MeSH
- RNA, Messenger analysis MeSH
- Polymerase Chain Reaction MeSH
- Rats, Inbred Lew MeSH
- Protein Isoforms MeSH
- Muscle Fibers, Slow-Twitch chemistry MeSH
- Muscle Fibers, Fast-Twitch chemistry MeSH
- Myosin Heavy Chains genetics MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Female MeSH
- Animals MeSH
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.
References provided by Crossref.org
Lit.: 26
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- $a Institute of Physiology, AS ČR, Prague
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- $a Lit.: 26
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- $a 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.
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