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Quantitative PCR analysis of laryngeal muscle fiber types
Journal article   Peer reviewed

Quantitative PCR analysis of laryngeal muscle fiber types

Douglas J Van Daele
Journal of communication disorders, Vol.43(4), pp.327-334
07/2010
DOI: 10.1016/j.jcomdis.2010.04.006
PMCID: PMC4530018
PMID: 20430402
url
http://doi.org/10.1016/j.jcomdis.2010.04.006View
Open Access

Abstract

Voice and swallowing dysfunction as a result of recurrent laryngeal nerve paralysis can be improved with vocal fold injections or laryngeal framework surgery. However, denervation atrophy can cause late-term clinical failure. A major determinant of skeletal muscle physiology is myosin heavy chain (MyHC) expression, and previous protein analyses have shown changes in laryngeal muscle fiber MyHC isoform with denervation. RNA analyses in this setting have not been performed, and understanding RNA levels will allow interventions better designed to reverse processes such as denervation in the future. Total RNA was extracted from bilateral rat thyroarytenoid (TA), posterior cricoarytenoid (PCA), and cricothyroid (CT) muscles in rats. Primers were designed using published MyHC isoform sequences. SYBR Green real-time reverse transcription-polymerase chain reaction (SYBR-RT-PCR) was used for quantification. The electropherogram showed a clear separation of total RNA to 28S and 18S subunits. Melting curves illustrated single peaks for all type MyHC primers. All MyHC isoforms were identified in all muscles with various degrees of expression. Quantitative PCR is a sensitive method to detect MyHC isoforms in laryngeal muscle. Isoform expression using mRNA analysis was similar to previous analyses but showed some important differences. This technique can be used to quantitatively assess response to interventions targeted to maintain muscle bulk after denervation. (1) Readers will be able to describe the relationship between myosin heavy chain expression and muscle contractile properties. (2) Readers will be able to separate myosin heavy chain isoforms into slow and fast twitch phenotypes. (3) Readers will be able to describe differential muscle isoform expression between different laryngeal muscles. (4) Readers will be able to compare this study to other modalities of determining muscle fiber type.
 Temperature Fluorescence Laryngeal Muscles - metabolism Myosin Heavy Chains - genetics Rats Muscle, Skeletal - metabolism RNA, Messenger - metabolism Myosin Heavy Chains - metabolism Rats, Sprague-Dawley Polymerase Chain Reaction - methods Animals Protein Isoforms - metabolism Time Factors Protein Isoforms - genetics

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