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Publication Date

2019

First Advisor

Stylianos P. Scordilis

Document Type

Honors Project

Degree Name

Bachelor of Arts

Department

Biochemistry

Keywords

Mice-Muscles, Proteomics, Myogenesis, Muscles, Satellite cells, C@C!@, Anabolic steroids, Anabolic-androgenic steroids, Nandrolone, Nandrolone decanoate, 19-nortestosteron, Skeletal muscle, Tandem mass tagging, Mass spectrometry

Abstract

Little is known about the biochemical mechanisms by which anabolic-androgenic steroids produce physiological changes in skeletal muscle tissue. In this study, C2C12 cells were grown in three different media: control (CTRL), vehicle control (DMSO), and anabolic steroid (nandrolone decanoate) containing (NAND). Cells were then harvested at three time points that represent the three stages of myogenesis: myoblasts (Day 0), early myotubes (Day 5) and late myotubes (Day 9). Whole cell extracts at each stage in each treatment (n=3) were labeled using tandem mass-tags (TMT) and combined into four separate TMT multiplex replicates. High-pressure liquid-chromatography-coupled mass spectrometry was used to identify and quantify the proteins expressed in each condition. Proteins determined to be myogenically-regulated in previous comprehensive literature on satellite cell myogenesis (Le Bihan et al., 2015) were selected and categorized into euKaryotic Orthologous Groups (KOGs) for functional annotation and analysis (Huerta Capas et al., 2016). KOGs that were significantly affected by NAND treatment compared to CTRL and DMSO treatments were further analyzed at the individual protein level. Proteins involved in cell cycle control, replication, and proliferation were found to decrease expression in the presence of nandrolone at the earlier stages of myogenesis, while those involved with cytoskeletal structure and strength, extracellular structure, membrane biogenesis, signal transduction, and energy production were found to increase expression in the presence of nandrolone during myogenesis. These proteomic results correlate with the reported physiological hypertrophic effects of anabolic-androgenic steroids reported in in vivo usage, and shed light on the biochemical mechanisms by which these molecules induce changes at the physiological level.

Rights

©2019 Emily Ann Morris. 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

viii, 142 pages : illustrations (some color). Includes bibliographical references (pages 134-142)

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