Publication Date

2013

Document Type

Masters Thesis

Department

Biological Sciences

Keywords

Mice-Muscles, Proteomics, Myogenesis, Muscles, Satellite cells, C2C12

Abstract

C2C12 muscle progenitor cells replicate, fuse and develop into spontaneously contracting muscle fibers in vitro, a process emulating the activity of satellite cells in vivo (Pannerec et al., 2012; Casadei et al., 2009). Proteomic analyses of these developing myocytes will shed light into the molecular mechanisms of myogenesis. Cells were harvested at three time-points representing three distinct stages of development: myoblasts (Day 0), early myotubes (Day 4) and late myotubes (Day 9). Whole cell extracts of each stage (n=5) were separated by two-dimensional SDS polyacrylamide gel electrophoresis and were analyzed to determine significant changes in protein expression. A combined total of 301 spots significantly changed intensity during development. Due to experimental limitations, 56 of these proteins were successfully identified and categorized into euKaryotic Orthologous Groups for functional annotation (Tatusov et al., 2003). Proteins generally involved in proliferation, protein synthesis and protein degradation were found to decrease expression steadily throughout myogenesis while proteins involved in energy production and sarcomere formation increased expression during this time. Proteins involved in cellular protection and protein stabilization such as molecular chaperones, antioxidants, and intermediate filament proteins either steadily increased expression during myogenesis or significantly increased expression during the myoblast to early myotube transition. Significant changes identified in the proteome during C2C12 myogenesis correlate with the shift in cellular morphology, physiology, and proteostasis of differentiation and create a more developed foundation for future research endeavors.

Language

English

Comments

ix, 51 pages : illustrations (some color) Thesis (M.S)--Smith College, 2013. Includes bibliographical references (pages 33-41)

Included in

Biology Commons

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