Summary
Although advancements in the preventive and therapeutic strategies of cardiac diseases have successfully improved the prognosis of many types of cardiac diseases, they are still challengeable targets because of their high mortality and large medical expenses. Moreover, because heart function is tightly associated with quality of life, it is important to elucidate the genetic and molecular basis of disease progression. One of the recent advances for assessing protein function is reverse chemical genetics, which has the advantages that complement classical reverse genetics and should advance efforts at drug discovery for many diseases. Toward that end an appropriate biological assay system is required to describe specific heart phenotypes.
Resent studies have shown that many aspects of Drosophila heart development and function are similar to those observed in the human heart, making Drosophila a useful model system with the advantage of a simpler genetic organization and shorter life span. Here we describe several assay systems that can be used to characterize Drosophila heart function. The first method is an external electrical pacing assay that is used to assess the response to stress in the adult fly. The incidence of pacing-induced heart dysfunction measured by this method strongly correlates with natural aging and mutation in genes known to be involved in human cardiac dysfunction. Consequently, this method can be used to identify unapparent heart failure phenotypes. This procedure is applicable for both genetic and pharmacological screening. The second method is an image-based heart performance assay. This method provides details of the dynamics of heart contraction in real time similar to clinical echocardiography. This method may be used for secondary drug screening as well as for more detailed analysis of the genetic and pharmacological phenotypes of Drosophila hearts.
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References
Brand AH, Perrimon N. (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401–15.
Kennerdell JR, Carthew RW. (2000) Heritable gene silencing in Drosophila using double-stranded RNA. Nat Biotechnol 18, 896–8.
Dietzl G, Chen D, Schnorrer F, et al. (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448,151–6.
Bilen J, Bonini NM. (2005) Drosophila as a model for human neurodegenerative disease. Annu Rev Genet 39, 153–71.
Marsh JL, Thompson LM. (2006) Drosophila in the study of neurodegenerative disease. Neuron 52, 169–78.
Bier E, Bodmer R. (2004) Drosophila, an emerging model for cardiac disease. Gene 342, 1–11.
Curtis NJ, Ringo JM, Dowse HB. (1999) Morphology of the pupal heart, adult heart, and associated tissues in the fruit fly, Drosophila melanogaster. J Morphol 240, 225–35.
Bodmer R, Venkatesh TV. (1998) Heart development in Drosophila and vertebrates: conservation of molecular mechanisms. Dev Genet 22, 181–6.
Cripps RM, Olson EN. (2002) Control of cardiac development by an evolutionarily conserved transcriptional network. Dev Biol 246, 14–28.
Olson EN. (2006) Gene regulatory networks in the evolution and development of the heart. Science 313, 1922–7.
Ocorr K, Perrin L, Lim HY, Qian L, Wu X, Bodmer R. (2007) Genetic control of heart function and aging in Drosophila. Trends Cardiovasc Med 17, 177–82.
Cammarato A, Dambacher CM, Knowles AF, et al. (2008) Myosin transducer mutations differentially affect motor function, myofibril structure, and the performance of skeletal and cardiac muscles. Mol Biol Cell 19, 553–62.
Paternostro G, Vignola C, Bartsch DU, Omens JH, McCulloch AD, Reed JC. (2001) Age-associated cardiac dysfunction in Drosophila melanogaster. Circ Res 88, 1053–8.
Wessells RJ, Bodmer R. (2004) Screening assays for heart function mutants in Drosophila. Biotechniques 37, 58–60, 62, 64 passim.
Wessells RJ, Fitzgerald E, Cypser JR, Tatar M, Bodmer R. (2004) Insulin regulation of heart function in aging fruit flies. Nat Genet 36, 1275–81.
Qian L, Liu J, Bodmer R. (2005) Neuromancer Tbx20-related genes (H15/midline) promote cell fate specification and morphogenesis of the Drosophila heart. Dev Biol 279, 509–24.
Akasaka T, Klinedinst S, Ocorr K, Bustamante EL, Kim SK, Bodmer R. (2006) The ATP-sensitive potassium (KATP) channel-encoded dSUR gene is required for Drosophila heart function and is regulated by tinman. Proc Natl Acad Sci USA 103, 11999–2004.
Peart JN, Gross GJ. (2002) Sarcolemmal and mitochondrial K(ATP) channels and myocardial ischemic preconditioning. J Cell Mol Med 6, 453–64.
Gross GJ, Fryer RM. (1999) Sarcolemmal versus mitochondrial ATP-sensitive K+ channels and myocardial preconditioning. Circ Res 84, 973–9.
Hanley PJ, Daut J. (2005) K(ATP) channels and preconditioning: A re-examination of the role of mitochondrial K(ATP) channels and an overview of alternative mechanisms. J Mol Cell Cardiol 39, 17–50.
Bienengraeber M, Olson TM, Selivanov VA, et al. (2004) ABCC9 mutations identified in human dilated cardiomyopathy disrupt catalytic KATP channel gating. Nat Genet 36, 382–7.
Ocorr K, Reeves NL, Wessells RJ, et al. (2007) KCNQ potassium channel mutations cause cardiac arrhythmias in Drosophila that mimic the effects of aging. Proc Natl Acad Sci USA 104, 3943–8.
Fink M, Callol-Massot C, Chu A, et al. (2009) A new method for Detection and Quantification of Heartbeat Parameters in Drosophila, Zebrafish, and Embryonic Mouse Hearts. BioTechniques 46, 101–113.
Wolf PA, Abbott RD, Kannel WB. (1991) Atrial fibrillation as an independent risk factor for stroke: the Framingham Study. Stroke 22, 983–8.
Lakatta EG, Levy D. (2003) Arterial and cardiac aging: major shareholders in cardiovascular disease enterprises: Part II: the aging heart in health: links to heart disease. Circulation 107, 346–54.
Olivetti G, Giordano G, Corradi D, et al. (1995) Gender differences and aging: effects on the human heart. J Am Coll Cardiol 26, 1068–79.
Acknowledgments
The authors would like to thank Professor Rolf Bodmer for his critical advice and encouragement. TA is a Sanford Fellow, supported by a fellowship of the Sanford Child Health Center at BIMR. KO is supported by a grant from the American Heart Association.
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Akasaka, T., Ocorr, K. (2009). Drug Discovery Through Functional Screening in the Drosophila Heart. In: Koga, H. (eds) Reverse Chemical Genetics. Methods in Molecular Biology™, vol 577. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-232-2_18
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DOI: https://doi.org/10.1007/978-1-60761-232-2_18
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