Endocrinol Metab.  2013 Jun;28(2):119-124. 10.3803/EnM.2013.28.2.119.

Expression of Thyroid Stimulating Hormone Receptor mRNA in Mouse C2C12 Skeletal Muscle Cells

• Ohn JH
• Han SK
• Park DJ
• Park KS
• Park YJ

• KMID: 1973810
• DOI: http://doi.org/10.3803/EnM.2013.28.2.119

Abstract

BACKGROUND
We analyzed whether thyroid stimulating hormone receptor (TSH-R) is expressed in a skeletal muscle cell line and if TSH has influence on the differentiation of muscle cells or on the determination of muscle fiber types.
METHODS
TSH-R gene expression was detected with nested real-time polymerase chain reaction (RT-PCR) in C2C12, a mouse skeletal muscle cell line. The effect of TSH on myotube differentiation was assessed by microscopic examination of myotube formation and through the measurement of expression of muscle differentiation markers, i.e., myogenin and myoD, and muscle type-specific genes, i.e., MyHC1, MyHC2a, and MyHC2b, with quantitative RT-PCR before and after incubation of C2C12 myotube with TSH.
RESULTS
TSH-R was expressed in the mouse skeletal muscle cell line. However, treatment with TSH had little effect on the differentiation of muscle cells, although the expression of the muscle differention marker myogenin was significantly increased after TSH treatment. Treatment of TSH did not affect the expression of muscle type-specific genes.
CONCLUSION
TSH-R is expressed in a mouse skeletal muscle cell line, but the role of TSH receptor signaling in skeletal muscle needs further investigation.

Keyword

Receptors, thyrotropin; Muscle, skeletal; Muscle differentiation

MeSH Terms

Animals
Antigens, Differentiation
Cell Line
Gene Expression
Mice
Muscle Cells
Muscle Fibers, Skeletal
Muscle, Skeletal
Muscles
Myogenin
Real-Time Polymerase Chain Reaction
Receptors, Thyrotropin
Thyroid Gland
Thyrotropin
Antigens, Differentiation
Myogenin
Receptors, Thyrotropin
Thyrotropin

Figure

• 

  Fig. 1 (A) Expression of thyroid stimulating hormone receptor (TSH-R) in mouse thyroid (positive control), skeletal muscle (gastrocnemius), heart, and small intestine (negative control) using real-time polymerase chain reaction (RT-PCR) detected with shorter primer sets used in nested RT-PCR (see Methods). TSH-R is expressed in skeletal muscle (gastrocnemius). (B) Expression of TSH-R in mouse hypothalamus (positive control) and C2C12 myoblast and myotube, detected with nested RT-PCR. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 2 Light microscopic images of C2C12 myotube after induction of differentiation with 2% horse serum without thyroid stimulating hormone (TSH) (A) or with TSH at concentrations of (B) 0.001, (C) 0.01, (D) 0.1, (E) 1, and (F) 10 mU/mL (×200).

• 

  Fig. 3 Expression of muscle differentiation markers, (A) myogenin and (B) myoD, in C2C12 myotube before and after treatment with thyroid stimulating hormone (TSH) and T3 for 24 hours at postdifferentiation day 5. The results are from three independent experiments. TSH and T3 significantly increase the expression of myogenin. aP<0.05.

• 

  Fig. 4 Expression of myosin heavy chain isoforms, (A) MyHC1, (B) MyHC2a, and (C) MyHC2b, in C2C12 myotube before and after treatment with thyroid stimulating hormone (TSH) and T3 for 24 hours at post-differentiation day 5. The result is from three independent experiments.

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