Author

Anagha Inguva

Publication Date

2014

Document Type

Honors Thesis

Department

Biochemistry

Keywords

Mice-Muscles, Carbohydrates-Metabolism, Myogenesis, Genetic transcription, Muscle contraction, Adenosine triphosphate, Glucose, C2C12, Carbohydrate, Metabolism, Murine, ATP, Contraction, Transcriptome

Abstract

Murine C2C12 cells, derived from adult dystrophic mouse thigh muscle simulate in vivo myogenesis (Yaffe and Saxel, 1977). They undergo three distinct stages of C2C12 myogenesis: proliferative myoblasts (day 0); cell cycle withdrawal and fusion to become early myotubes (day 4); and, finally spontaneously contracting late myotubes (day 9). The present study analyzed the changes in the transcriptome of the enzymes involved in carbohydrate metabolism during the three different stages of myogenic development of C2C12 cells. A carbohydrate metabolism qRT-PCR array was used to examine the mRNA levels of 84 different enzymes involved in carbohydrate metabolism (n=4 per stage). A total of 64 genes showed changes in mRNA levels that were statistically significant (ANOVA, p<0.05) from the control group (day 0). Of these 64 genes, 29 genes increased or decreased by two (2.0) fold or higher and were considered to be biologically significant changes. Eleven genes were involved in the TCA cycle, 5 genes were involved in glycolysis, 7 genes were involved in glycogen metabolism, 4 genes were involved in the pentose phosphate pathway and two genes were involved in gluconeogenesis. Isoenzyme mRNA level shifts were also present during the three stages of myogenesis. In addition, the enzymatic activities of citrate synthase (CS) and creatine kinase (CK) were measured (n=6) during each stage. CS activity was 112, 204, and 271 nmol/min/mg total protein and 37, 62 and 105 nmol/min/mg total DNA respective to day 0, 4, and 9. CK activity was 86, 342, and 532 nmol/min/mg total protein and 29, 105, 661 nmol/min/mg total DNA respective to day 0, 4, and 9. These data indicate that the TCA cycle, glycolysis and glycogen metabolism pathways were significantly altered during myogenesis to meet the energetic and synthetic needs of developing cells.

Language

English

Comments

ix, 104 pages : illustrations (some color). Honors Project-Smith College, 2014. Includes bibliographical references (pages 99-104)

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