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Oigo FINAL 12 14 2021 with cert (1).pdf (1.31 MB)

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Gene Expression in Long-term myoblast/myocyte cultures: RNA Analysis (DYSTROPHIN GENE)

Oigo, Annah Bochaberi
http://orcid.org/0000-0002-1479-0184
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1640077186019624

Abstract Details

2021, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
Myogenesis involves changes in gene expression which is necessary for muscle development and regeneration. This involves the fusion of myoblasts to form myotubes which mature into myofibers to form new skeletal muscles (important in the movement of the body). This process is regulated by Myogenesis Regulatory Factors (MRFs) which include MyoD, Myogenin, Myf5 and MRF4. This study examines how different genes are expressed when myoblast C2C12 cells are grown in long term cultures. We conducted an experimental study design using the immortalized mouse myoblast cell line C2C12. The cells were allowed to differentiate and were harvested at designated time points, seven time points. The focus of this study was to look at expression of dystrophin gene throughout the process of myogenesis. The C2C12 cells were grown and allowed to differentiate over a 30-day time course study with seeding, imaging, growth media change (10% FBS), and pelleting cells at specific time points for RNA isolation. Following the RNeasy kit protocol the RNA was isolated and quantified by nanodrop spectrophotometry. Quantitative Real-Time Polymerase Chain Reaction (QRT-PCR) analysis was carried out to measure expression of dystrophin gene. Gene expression analysis was normalized to GAPDH. Expression of dystrophin gene was measured for the two trials and later the average was done. Dystrophin expression levels are high during activation of satellite cells where it helps to regulate cell polarity and increase number of asymmetric divisions. As the activated cells proliferate, dystrophin levels go down. After the onset of differentiation, increased levels of dystrophin expression are observed which help in plasma membrane stabilization of striated muscle cells. During muscle aging, expression levels slightly decrease to maintain the stability of the cell membrane.
Gary Walker, PhD (Advisor)
Jonathan Caguiat, PhD (Committee Member)
David Asch, PhD (Committee Member)
70 p.
Biology; Cellular Biology; Genetics; Molecular Biology
Dystrophin; Skeletal muscle; Myogenesis; Muscle aging

Recommended Citations

Citations

  • Oigo, A. B. (2021). Gene Expression in Long-term myoblast/myocyte cultures: RNA Analysis (DYSTROPHIN GENE) [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1640077186019624

    APA Style (7th edition)

  • Oigo, Annah. Gene Expression in Long-term myoblast/myocyte cultures: RNA Analysis (DYSTROPHIN GENE). 2021. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1640077186019624.

    MLA Style (8th edition)

  • Oigo, Annah. "Gene Expression in Long-term myoblast/myocyte cultures: RNA Analysis (DYSTROPHIN GENE)." Master's thesis, Youngstown State University, 2021. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1640077186019624

    Chicago Manual of Style (17th edition)

ysu1640077186019624
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© 2021, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.