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Publication Date

2022-8

First Advisor

Stylianos P. Scordilis

Document Type

Masters Thesis

Degree Name

Master of Science

Department

Biological Sciences

Keywords

Sex Hormone Receptor, Skeletal Muscle, SHR Modulator Treatment, Signal Protein Phosphorylation, C2C12 Myogenesis

Abstract

The goal of this experiment is to explore the role of sex hormone receptor (SHR) activity in the development of skeletal muscle. Murine myoblast cells from the C2C12 line were cultured in eight experimental conditions with an agonized and antagonized state for each of the four SHRs. Cell morphology, changes in viability, and proliferation rates were analyzed for all treatments. Total protein concentrations of each cell population were estimated via the Lowry Assay and specific proteins were identified with gel electrophoresis. Immunoblotting was used to confirm the presence of the SHRs and key signaling proteins, and to identify changes in their relative proportions. The following signaling proteins were analyzed: p38α , ERK1/2, Akt/PKB, and JNK 1/2. ImageJ was used to quantify the band intensities on the gels and blots. The activation ratios of nonphosphorylated to phosphorylated protein isoforms was determined under each experimental condition to ascertain their regulation. Significant dose-dependent correlations were seen between SHR modulator treatment and the following variables: viability/proliferation rate, total protein concentration/protein mass per cell, receptor expression, Akt/pAkt expression, ERK/pERK expression, and p38α /pp38α expression. The effects of SHR modulator treatment were particularly evident in the final stage of myogenesis (day 9). Interesting results were observed regrading differential expression of GPER proteoforms during different stages of myogenesis. Together, these results suggest that sex hormone receptors do play a significant role in C2C12 myogenesis.

Rights

©2022 Ilemona Ameadaji. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

Comments

126 pages: illustrations (some color). Includes bibliographical references (pages 106 -126).

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