Glucocorticoid Inhibition of C2C12 Proliferation Rate and Differentiation Capacity in Relation to MRNA Levels of the MRF Gene Family

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2000 Nov 25

PMID 11092555

Citations 25

Authors

M F te Pas

P R de Jong

F J Verburg

Affiliations

Soon will be listed here.

Abstract

The muscle regulatory factors (MRF) gene family regulate muscle fibre development. Several hormones and drugs also affect muscle development. Glucocorticoids are the only drugs reported to have a beneficial effect on muscle degenerative disorders. We investigated the glucocorticoid-related effects on C2C12 myoblast proliferation rate, morphological differentiation, and subsequent mRNA expression patterns of the MRF genes. C2C12 cells were incubated with the glucocorticoids dexamethasone or alpha-methyl-prednisolone. Both glucocorticoids showed comparable effects. Glucocorticoid treatment of C2C12 cells during the proliferative phase reduced the proliferation rate of the cells dose dependently, especially during the third and fourth day of culture, increased MyoD1, myf-5, and MRF4 mRNA levels, and reduced myogenin mRNA level, compared to untreated control cells. Thus, the mRNA level of proliferation-specific MyoD1 and myf-5 expression does not seem to associate with C2C12 myoblast proliferation rate. Glucocorticoid treatment of C2C12 cells during differentiation reduced the differentiation capacity dose dependently, which is accompanied by a dose dependent reduction of myogenin mRNA level, and increased MyoD1, myf-5, and MRF4 mRNA levels compared to untreated control cells. Therefore, we conclude that glucocorticoid treatment reduces differentiation of C2C12 myoblasts probably through reduction of differentiation-specific myogenin mRNA level, while inducing higher mRNA levels of proliferation-associated MRF genes.

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References

Edmondson D, Cheng T, Cserjesi P, Chakraborty T, Olson E . Analysis of the myogenin promoter reveals an indirect pathway for positive autoregulation mediated by the muscle-specific enhancer factor MEF-2. Mol Cell Biol. 1992; 12(9):3665-77. PMC: 360220. DOI: 10.1128/mcb.12.9.3665-3677.1992. View

Guerriero Jr V, FLORINI J . Dexamethasone effects on myoblast proliferation and differentiation. Endocrinology. 1980; 106(4):1198-202. DOI: 10.1210/endo-106-4-1198. View

Passaquin A, Lhote P, Ruegg U . Calcium influx inhibition by steroids and analogs in C2C12 skeletal muscle cells. Br J Pharmacol. 1998; 124(8):1751-9. PMC: 1565578. DOI: 10.1038/sj.bjp.0702036. View

Carnac G, Levin A, Bonnieu A . 9-cis-retinoic acid regulates the expression of the muscle determination gene Myf5. Endocrinology. 1993; 133(5):2171-6. DOI: 10.1210/endo.133.5.8404668. View

Anderson J, McIntosh L, Poettcker R . Deflazacort but not prednisone improves both muscle repair and fiber growth in diaphragm and limb muscle in vivo in the mdx dystrophic mouse. Muscle Nerve. 1996; 19(12):1576-85. DOI: 10.1002/(SICI)1097-4598(199612)19:12<1576::AID-MUS7>3.0.CO;2-7. View

Beilharz M, Lareu R, Garrett K, Grounds M, Fletcher S . Quantitation of muscle precursor cell activity in skeletal muscle by Northern analysis of MyoD and myogenin expression: Application to dystrophic (mdx) mouse muscle. Mol Cell Neurosci. 2009; 3(4):326-31. DOI: 10.1016/1044-7431(92)90029-2. View

te Pas M, Verburg F, Gerritsen C, de Greef K . Messenger ribonucleic acid expression of the MyoD gene family in muscle tissue at slaughter in relation to selection for porcine growth rate. J Anim Sci. 2000; 78(1):69-77. DOI: 10.2527/2000.78169x. View

Aurade F, Pfarr C, Lindon C, Garcia A, Primig M, Montarras D . The glucocorticoid receptor and AP-1 are involved in a positive regulation of the muscle regulatory gene myf5 in cultured myoblasts. J Cell Sci. 1998; 110 ( Pt 22):2771-9. DOI: 10.1242/jcs.110.22.2771. View

Olson E . MyoD family: a paradigm for development?. Genes Dev. 1990; 4(9):1454-61. DOI: 10.1101/gad.4.9.1454. View

10.

Weintraub H, Davis R, Tapscott S, Thayer M, Krause M, Benezra R . The myoD gene family: nodal point during specification of the muscle cell lineage. Science. 1991; 251(4995):761-6. DOI: 10.1126/science.1846704. View

11.

Tsai C, Saffitz J, Billadello J . Expression of the Gs protein alpha-subunit disrupts the normal program of differentiation in cultured murine myogenic cells. J Clin Invest. 1997; 99(1):67-76. PMC: 507769. DOI: 10.1172/JCI119135. View

12.

Giorgino F, Smith R . Dexamethasone enhances insulin-like growth factor-I effects on skeletal muscle cell proliferation. Role of specific intracellular signaling pathways. J Clin Invest. 1995; 96(3):1473-83. PMC: 185771. DOI: 10.1172/JCI118184. View

13.

van den Hemel-Grooten H, te Pas M, van den Bosch T, Garssen G, Schreurs V, Verstegen M . mRNA levels of the calpain system in longissimus muscle of young pigs during prolonged feeding of a protein-free diet. J Anim Sci. 1997; 75(4):968-74. DOI: 10.2527/1997.754968x. View

14.

Koishi K, Zhang M, McLennan I, Harris A . MyoD protein accumulates in satellite cells and is neurally regulated in regenerating myotubes and skeletal muscle fibers. Dev Dyn. 1995; 202(3):244-54. DOI: 10.1002/aja.1002020304. View

15.

Montano M, Lim R . Glucocorticoid effects on the skeletal muscle differentiation program: analysis of clonal proliferation, morphological differentiation and the expression of muscle-specific and regulatory genes. Endocr Res. 1997; 23(1-2):37-57. DOI: 10.1080/07435809709031841. View

16.

Chomczynski P, Sacchi N . Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987; 162(1):156-9. DOI: 10.1006/abio.1987.9999. View

17.

Desler M, Jones S, Smith C, Woods T . Effects of dexamethasone and anabolic agents on proliferation and protein synthesis and degradation in C2C12 myogenic cells. J Anim Sci. 1996; 74(6):1265-73. DOI: 10.2527/1996.7461265x. View

18.

Grounds M, Garrett K, Lai M, Wright W, Beilharz M . Identification of skeletal muscle precursor cells in vivo by use of MyoD1 and myogenin probes. Cell Tissue Res. 1992; 267(1):99-104. DOI: 10.1007/BF00318695. View

19.

Fuchtbauer E, Westphal H . MyoD and myogenin are coexpressed in regenerating skeletal muscle of the mouse. Dev Dyn. 1992; 193(1):34-9. DOI: 10.1002/aja.1001930106. View

20.

Mendell J, Moxley R, Griggs R, Brooke M, Fenichel G, Miller J . Randomized, double-blind six-month trial of prednisone in Duchenne's muscular dystrophy. N Engl J Med. 1989; 320(24):1592-7. DOI: 10.1056/NEJM198906153202405. View