Quantification of myosin heavy chain mRNA in somatic and branchial arch muscles using competitive PCR

Hak Hyun Jung, Richard L. Lieber, Allen F. Ryan

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The purpose of this study was to quantify the type and amount of myosin heavy chain (MHC) mRNA within muscles of different developmental origins to determine whether the regulation of gene expression is comparable. Seven MHC isoforms were analyzed in rat adult limb (extensor digitorum longus, tibialis anterior, and soleus) and nonlimb (extraocular, thyroarytenoid, diaphragm, and masseter) muscles using a competitive PCR assay. An exogenous template that included oligonucleotide sequences specific for seven rat sarcomeric MHC isoforms (β-cardiac, 2A, 2X, 2B, extraocular, embryonic, and neonatal) as well as β-actin was constructed and used as the competitor. Only the extraocular muscle contained all seven isoforms. All seven muscles contained type 2A and type 2X MHC transcripts in varying percentages. As expected, the soleus muscle contained primarily β-cardiac MHC (87.8 ± 2.6%). Extraocular MHC was found only in the extraocular and thyroarytenoid muscles and in relatively small proportions (7.4 ± 1.5% and 4.0 ± 0.7%, respectively). Neonatal MHC was identified in extraocular (7.9 ± 0.3%), thyroarytenoid (4.4 ± 0.4%), and masseter (1.0 ± 0.2%) muscles, and embryonic MHC was identified both in extraocular (1.2 ± 0.5%) and, unexpectedly, in soleus (0.6 ± 0.1%) muscles. Absolute MHC mRNA mass was greatest in the masseter (106 pg/0.5/μg RNA) and least for the tibialis anterior (64 pg/0.5 μg RNA). These values suggest that MHC mRNA represents from 4 to 17% of the total mRNA pool in various skeletal muscles. Differences in MHC profile between somatic and branchial arch muscles suggest that the developmental origin of a muscle may, at least in part, be responsible for the MHC expression program that is implemented in the adult. An inverse relationship between the expression of β-cardiac and type 2B MHC transcripts across muscles was noted, suggesting that the expression of these two isoforms may be reciprocally regulated.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume275
Issue number1 44-1
Publication statusPublished - 1998 Jul 1

Fingerprint

Branchial Region
Myosin Heavy Chains
Muscles
Polymerase Chain Reaction
Messenger RNA
Oculomotor Muscles
Protein Isoforms
Cardiac Myosins
Skeletal Muscle
RNA
Laryngeal Muscles
Masseter Muscle
Gene Expression Regulation
Diaphragm
Oligonucleotides

Keywords

  • Gene expression
  • Polymerase chain reaction
  • Rat

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

Quantification of myosin heavy chain mRNA in somatic and branchial arch muscles using competitive PCR. / Jung, Hak Hyun; Lieber, Richard L.; Ryan, Allen F.

In: American Journal of Physiology - Cell Physiology, Vol. 275, No. 1 44-1, 01.07.1998.

Research output: Contribution to journalArticle

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abstract = "The purpose of this study was to quantify the type and amount of myosin heavy chain (MHC) mRNA within muscles of different developmental origins to determine whether the regulation of gene expression is comparable. Seven MHC isoforms were analyzed in rat adult limb (extensor digitorum longus, tibialis anterior, and soleus) and nonlimb (extraocular, thyroarytenoid, diaphragm, and masseter) muscles using a competitive PCR assay. An exogenous template that included oligonucleotide sequences specific for seven rat sarcomeric MHC isoforms (β-cardiac, 2A, 2X, 2B, extraocular, embryonic, and neonatal) as well as β-actin was constructed and used as the competitor. Only the extraocular muscle contained all seven isoforms. All seven muscles contained type 2A and type 2X MHC transcripts in varying percentages. As expected, the soleus muscle contained primarily β-cardiac MHC (87.8 ± 2.6{\%}). Extraocular MHC was found only in the extraocular and thyroarytenoid muscles and in relatively small proportions (7.4 ± 1.5{\%} and 4.0 ± 0.7{\%}, respectively). Neonatal MHC was identified in extraocular (7.9 ± 0.3{\%}), thyroarytenoid (4.4 ± 0.4{\%}), and masseter (1.0 ± 0.2{\%}) muscles, and embryonic MHC was identified both in extraocular (1.2 ± 0.5{\%}) and, unexpectedly, in soleus (0.6 ± 0.1{\%}) muscles. Absolute MHC mRNA mass was greatest in the masseter (106 pg/0.5/μg RNA) and least for the tibialis anterior (64 pg/0.5 μg RNA). These values suggest that MHC mRNA represents from 4 to 17{\%} of the total mRNA pool in various skeletal muscles. Differences in MHC profile between somatic and branchial arch muscles suggest that the developmental origin of a muscle may, at least in part, be responsible for the MHC expression program that is implemented in the adult. An inverse relationship between the expression of β-cardiac and type 2B MHC transcripts across muscles was noted, suggesting that the expression of these two isoforms may be reciprocally regulated.",
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