Prenatal alcohol exposure causes a range of developmental disorders that are commonly referred to as fetal alcohol spectrum disorders (FASDs). The most severe case of FASDs is known as fetal alcohol syndrome (FAS) and is characterized by growth deficiencies, craniofacial malformations, and both structural and functional central nervous system (CNS) abnormalities. The combination of technological limitations that decrease experimental throughput and the inherent stochasticity of biological processes eliminate the ability to gain a quantitative understanding about the onset and progression of ethanol-related developmental disorders. As a result there is not much known mechanistically about how ethanol exposure manifests as FASDs. Here, we aim to develop a microfluidic-based system that enables high-throughput and high-content studies to elucidate the effects of ethanol exposure on embryogenesis. The model organism Drosophila melanogaster (Drosophila) has been shown to be a model of FASDs, because Drosophila exhibits similar phenotypical defects as humans when embryos are raised on ethanol solutions. It has yet to be seen how ethanol exposure affects developmental processes that occur during embryogenesis that lead to the wide ranging symptoms of FASDs. The technological gap that precedes this is a method that can robustly expose developing fly embryos to ethanol while also allowing high-throughput live imaging and data acquisition. The objective of this project is to develop enabling technologies for whole embryo imaging to uncover the mechanisms through which embryonic ethanol exposure affects morphogenesis and developmental patterning that ultimately lead to FASDs. In order to address the technological gap we are developing a microfluidic system that can culture and expose live Drosophila embryos to ethanol in a highly-parallelized manner. This system will also enable live imaging of embryogenesis, so that we can better understand how ethanol exposure affects morphogenesis. Furthermore, the microfluidic system will be integrated with tissue clearing techniques that will enable whole embryo imaging and thus allow us to visualize whole embryo gene expression patterns. In this study, we will study mutants of key enzymes to better elucidate their roles in FASD. Using this device, I aim to better our understanding about the physical and gene expression changes involved with FASDs. Microfluidic systems will be integrated with computer automation programs that will increase data acquisition rates. Data analysis will be streamlined through the development of computer vision algorithms that will help identify phenotypical changes associated with ethanol exposure that are non-obvious and likely to be missed by manual annotation. This project will not only enlarge the repertoire of tools available for developmental biology in general, but also will have significant impact on ethanol teratology.
The purpose of this project is to develop a high-throughput, microfluidics-based system for studying the physical and genetic origins of fetal alcohol spectrum disorders (FASDs). The results of this project will greatly increase our understanding of how prenatal ethanol exposure affects early embryogenesis and elucidate the mechanism through which ethanol exposure results in a wide range of developmental defects. Insights gained from this project will lead to novel treatments to help reduce ethanol teratogenicity and prevent the onset of FASDs.
|Levario, Thomas J; Zhao, Charles; Rouse, Tel et al. (2016) An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos. Sci Rep 6:21366|