White Rose University Consortium logo
University of Leeds logo University of Sheffield logo York University logo

The role of the transcription factor klf2a in vascular biology

Novodvorsky, Peter (2014) The role of the transcription factor klf2a in vascular biology. PhD thesis, University of Sheffield.

[img]
Preview
Text
Thesis The role of the transcription factor klf2a in vascular biology.pdf
Available under License Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 UK: England & Wales.

Download (4Mb) | Preview

Abstract

Introduction: The zinc-finger transcription factor Krϋppel-like factor 2 (KLF2) transduces physical forces of blood flow into molecular signals responsible for a wide range of biological responses. KLF2 maintains a healthy, quiescent endothelial phenotype. I studied the expression and function of the zebrafish KLF2 ortholog klf2a in vascular biology. Materials and Methods: Expression patterns of genes were observed using Whole- mount in situ hybridisation (WISH) technique. Relative expressions of genes were investigated using reverse transcription quantitative polymerase chain reaction (RT- qPCR). Particle imaging velocimetry (PIV) method was used to measure blood flow velocities. Blood flow in zebrafish embryos was manipulated genetically and pharmacologically. Transcription Activator-Like Effector Nucleases (TALEN) were used to generate a stable klf2a mutant line. Western blot, mass spectrometry and immunoprecipitation techniques were used in Klf2a protein studies. klf2a mutant lines were crossed with several transgenic reporter lines to study the role of klf2a in vascular development and transcription. Results: I reproduced and extended previous studies of spatial and temporal klf2a expression patterns. I confirmed that this expression is dependent on blood flow. Morpholino-mediated klf2a knockdown was used to study the roles of klf2a in regulation of other previously identified flow-dependent genes. klf2a mutants were generated and phenotyped. The mutant phenotype was compared to the one seen in klf2a MO morphants.

Item Type: Thesis (PhD)
Keywords: klf2a, blood-flow, shear stress, mutant
Academic Units: The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > Medicine (Sheffield)
Identification Number/EthosID: uk.bl.ethos.632982
Depositing User: Dr Peter Novodvorsky
Date Deposited: 14 Jan 2015 15:00
Last Modified: 12 Oct 2018 09:21
URI: http://etheses.whiterose.ac.uk/id/eprint/7697

You do not need to contact us to get a copy of this thesis. Please use the 'Download' link(s) above to get a copy.
You can contact us about this thesis. If you need to make a general enquiry, please see the Contact us page.

Actions (repository staff only: login required)