Development of a Voltage Sensor Tool, mermaid-2, for Use During Fertilization in Caenorhabditis elegans Open Access

Bastien, Amanda Joy (2015)

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My lab studies Caenorhabditis elegans, which is a great model organism for studying fertilization. Many different fertilization defective mutants have been identified and now there is a need for new tools to analyze the phenotoypes of these mutants. In particular, we are interested in studying the spe-9 class of mutants. The spe-9 class includes mutants in eight genes, all of which are able to complete spermatogenesis and appear indistinguishable from wild type, except that they cannot fertilize an oocyte when they contact it. All eight genes cause the same fertilization-defective phenotype and, currently, there is no way to phenotypically distinguish them from each other. My goal was to develop a voltage sensor tool to use in vivo in Caenorhabditis elegans to help determine if there is a voltage change during fertilization. Previously, fluorescent reporters have been used in a wide variety of organisms to measure voltage changes in vivo (Tsutsui et al. 2008; 2013). My thesis is centered on creating a mermaid-2 gene that could be used in the Caenorhabditis elegans germline, microinjecting the plasmid, confirming its presence, and detecting mermaid-2 expression. However, the expression of mermaid-2 is still being investigated in current transgenic lines.

Table of Contents

Chapter I: General Introduction

Chaper II: Introduction to Caenorhabditis elegans

Chaper III: Introduction to Project

Chapter IV: Methodology

Chapter V: Results

Chapter VI: Discussion

Chapter VII: Next Steps

Chapter VIII: Big Picture

Chapter IX: Figures & Pictures

Chapter X: Supplemental Information

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