Epigenetic Control of Skeletal Muscle Development: Identification and Characterization of Regulatory Elements

Smith, Emily (2021) Epigenetic Control of Skeletal Muscle Development: Identification and Characterization of Regulatory Elements. Masters thesis, University of East Anglia.

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Abstract

The musculoskeletal system is a crucial component for most vertebrate organisms that rely on movement. During development, the musculoskeletal system arises from clusters of mesoderm, which form bilaterally to the notochord, termed somites. Somites contain the cellular precursors of skeletal muscle, tendons, cartilage, some bone, and the dermis. Myogenesis, chondrogenesis and the development of tendons are all controlled by gene regulatory networks, that allow for the proliferation, specification, and differentiation of different cell lineages. The activity of these genes can be regulated through epigenetic mechanisms, including the activity of cis-regulatory regions of DNA known as enhancers. The aim of this research project was to characterise genes involved with myogenesis, chondrogenesis and the development of tendons and identify any novel enhancers that are regulating these genes. Enhancers are found in regions of open chromatin, to allow for transcription factor binding. ATAC-sequencing (Assay for Transposase Accessible Chromatin) identifies these regions of open chromatin and a dataset has been previously generated in the lab from somites in stage 14 chicken embryos. This data was utilised to find putative enhancer regions. These identified candidate enhancer regions were cloned into a fluorescent reporter construct which was injected and electroporated into HH3+ chicken embryos. Genes that had yet to be characterised in the chicken embryo were visualised spatially and temporally throughout development using whole mount in-situ hybridisation techniques, generating gene expression profiles. An expression profile for the fork-head protein Foxo1 was generated using in-situ hybridisation and cryosectioning techniques, along with the identification of the Scleraxis gene in younger embryos than previously reported. One enhancer associated with the Foxo1 gene showed fluorescent reporter activity in blood islands and sporadically throughout the inter-somitic blood vessels and the dorsal aorta.

Item Type:	Thesis (Masters)
Faculty \ School:	Faculty of Science > School of Biological Sciences
Depositing User:	Chris White
Date Deposited:	25 Aug 2021 13:20
Last Modified:	25 Aug 2021 13:20
URI:	https://ueaeprints.uea.ac.uk/id/eprint/81212
DOI:

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