High-throughput Whole Ganglion Neural Imaging in Caenorhabditis elegans with Microfluidic Systems Open Access

Abstract

The relationship between neural structure and function at a systemic level is important in understanding how disease affects the brain. Investigation of such implicates pathophysiology of diseases, including neurodevelopmental disorders such as autism spectrum disorder (ASD) and neurodegenerative diseases such as Alzheimer's Disease (AD) and Huntington Disease (HD). Although higher organisms, including mice, Rhesus Macaques, and humans, are very valuable in understanding brain function, the scale and scope of such studies is limited for ethical reasons as well as by monetary resources. In comparison, Caenorhabditis elegans is a small nematode that can be cultured isogenically in bulk. It is the only animal model whose nervous system structure has been completely mapped, making localization of function possible. It is also the only organism in which the entire head ganglion, can be simultaneously imaged, capturing different classes of neurons including sensory, motor, and interneurons. This presents a unique opportunity for complete mapping of the relationship between neural structure and function. However, traditional data acquisition and processing approaches are low-throughput, presenting challenges for efficient whole ganglion neural analysis. This thesis thus presents preliminary data enabled by microfluidic devices, introducing an effective method to increase throughput for investigation on neural structure and function.

We interrogate whole ganglion neural properties from two fronts:1) functional imaging of neurons and 2) static imaging of synapses. For functional imaging, we increased efficiency at both the data acquisition and processing level by acquiring large-scale data with the help of microfluidic devices and through the validation of a developed 3D neuronal tracker. For static imaging, we qualitatively phenotyped the synapses of worms with developmental and genotypic variations. We then quantitatively compared the heterogeneity of neural features that contributes to the formation of a mature functional nervous system through an image processing and statistical analysis pipeline. The two parallel studies highlight the significance of whole ganglion neural imaging in C. elegans enabled by microfluidic systems. These initial studies demonstrate the potential for this methodology to inform our understanding of variability in neural structure and function at a systemic level.

Table of Contents

Table of Contents

Abstract............................................................................................................................................1
Introduction......................................................................................................................................3
Methods..........................................................................................................................................12

Table 1 ............................................................................................................................16

Results............................................................................................................................................19

Figure 1 ..........................................................................................................................19

Figure 2 ..........................................................................................................................20

Figure 3 ..........................................................................................................................23

Figure 4 ..........................................................................................................................28

Figure 5 ..........................................................................................................................34

Discussion & Conclusion....................................................................................................................37

References......................................................................................................................................45

Appendices.....................................................................................................................................49

Figure S1 ........................................................................................................................49

Figure S2 ........................................................................................................................50

Figure S3 ........................................................................................................................51

Figure S4 ........................................................................................................................51

Figure S5 ........................................................................................................................52

About this Honors Thesis

Rights statement

• Permission granted by the author to include this thesis or dissertation in this repository. All rights reserved by the author. Please contact the author for information regarding the reproduction and use of this thesis or dissertation.

School	Emory College
Department	Neuroscience and Behavioral Biology
Degree	BS
Submission	Honors Thesis
Language	English
Research Field	Engineering, Biomedical Biology, Neuroscience
Keyword	Static Imaging Image Processing Brain-wide sampling Functional Imaging Microfluidic Systems Caenorhabditis elegans Whole Ganglion Neural Analysis Neuronal Tracker
Committee Chair / Thesis Advisor	Lu, Hang, School of Chemical and Biomolecular Engineering, GA Tech Prinz, Astrid A, Emory University
Committee Members	Bates, Kathleen, School of Chemical and Biomolecular Engineering, GA Tech Crutcher, Michael D, Emory University Raper, Jessica, Emory University

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