Hartley, Paul

Heck, Margarete

Catterson, James Harold

2014-05-02T13:07:58Z

2014-05-02T13:07:58Z

2013-11-29

Background/Aims. Understanding the fundamental mechanisms underlying the development of congenital heart disease and cardiomyopathies is a goal of researchers worldwide, with the ultimate goal being the establishment of effective therapeutics for the amelioration of cardiac dysfunction. Unfortunately these disorders are often polygenic in aetiology, making it difficult for researchers to probe complex interactions that may contribute to the severity of the disease. Over the last decade, the adult fruit fly (Drosophila melanogaster) has emerged as an invaluable tool with which to study the genetic and molecular mechanisms underlying heart function. The aim of my thesis research was to establish the adult fruit fly as a model of human heart function, and to exploit this powerful genetic system to screen for conserved genes affecting the development and function of its cardiac syncytium. Methodology/Results Baseline measures of heart function and other factors contributing to variability in heart function (i.e. age, temperature, and the time of day) were assessed to establish the adult Drosophila heart model. I then performed an a priori RNAi screen, knocking down expression of individual conserved genes via cardiomyocyte-specific overexpression utilising the yeast GAL4/UAS system. Heart-specific ablation of Fermitin 1 and Fermitin 2 (Fit1, Fit2), the two Drosophila orthologs of Kindlin 2 (Kind2, a gene thought to be important for cardiomyocyte-cardiomyocyte junction integrity in human myocardium), caused severe cardiomyopathy characterised by the failure of cardiomyocytes to develop as a functional syncytium and loss of synchrony between cardiomyocytes. I generated a null allele of Fit1 via P-element mobilisation, but this had no impact on heart development or function. Similarly, the silencing of Fit2 failed to affect heart development or function. In contrast, the silencing of Fit2 in the cardiomyocytes of Fit1-null flies disrupted syncytium development, leading to severe cardiomyopathy. Temperature-sensitive cardiac-specific GAL4/GAL80ts lines were also generated, and knockdown of Fit (Fit1 and Fit2) function at different developmental stages was assessed. I observed the strongest effects of Fit knockdown on adult cardiac morphology during stages of heart development and remodelling, with significant cardiomyocyte decoupling. After 3-weeks of Fit knockdown during adulthood, cardiomyocytes were significantly decoupled, and these hearts were significantly arrhythmic compared to control animals. Conclusions/Discussion. My data provide clarity about the role of Kind2 by demonstrating a cell autonomous role for this family in the development of a functional cardiac syncytium in Drosophila. My findings also show that the Fermitins can functionally compensate for each other in order to control syncytium development. Therefore, my thesis demonstrates the power of the fruit fly as a model of human cardiac physiology, and supports the concept that abnormalities in cardiomyocyte KIND2 expression or function may contribute to cardiomyopathies in humans.

http://hdl.handle.net/1842/8802

en

The University of Edinburgh

Fermitins, the orthologs of mammalian Kindlins, regulate the development of a functional cardiac syncytium in Drosophila melanogaster. Catterson JH, Heck MMS, Hartley PS. PLoS One. 2013 May 15;8(5):e62958.

Diurnal expression of the thrombopoietin gene is regulated by CLOCK. Tracey CJ, Pan X, Catterson JH, Harmar AJ, Hussain MM, Hartley PS. J Thromb Haemost. 2012 Apr;10(4):662-9.

Cyclophilin A is a damage-associated molecular pattern molecule that mediates acetaminophen-induced liver injury. Dear JW, Simpson KJ, Nicolai MP, Catterson JH, Street J, Huizinga T, Craig DG, Dhaliwal K, Webb S, Bateman DN, Webb DJ. J Immunol. 2011 Sep 15;187(6):3347-52.

Dietary modulation of Drosophila sleep-wake behaviour. Catterson JH, Knowles-Barley S, James K, Heck MM, Harmar AJ, Hartley PS. PLoS One. 2010 Aug 10;5(8):e12062.

Drosophila

Heart

Establishing Drosophila as a model to study the functional relevance of conserved heart genes

Thesis or Dissertation

Doctoral

PhD Doctor of Philosophy