Title
Optimizing Tools for the Investigation of Flagella Protein Trafficking in Chlamydomonas Reinhardtii
Date of Award
Spring 2019
Document Type
Thesis
Degree Name
Master of Arts (MA)
Department
Biological Sciences
First Advisor
Yang, Pinfen
Second Advisor
Abbott, Allison
Third Advisor
Stuart, Rosemary
Abstract
Cilia and flagella are synonymous organelles that protrude above the cell surface to propel fluid and monitor signals in the environment. These functions are central to human development and health, as well as for numerous organisms to thrive in their natural habitats. Pioneering studies of their fundamental mechanisms heavily rely on the biflagellate green alga, Chlamydomonas. One example is the extensive characterization of bidirectional intraflagellar transport (IFT), a transport system that perpetually delivers flagellar components between the base and the tip of the flagella. Such characterization has resulted in the discovery of numerous evolutionarily conserved genes crucial for flagella growth and function. Yet, intense autofluorescence emitted mostly from photosynthesis pigments has prevented visualization of flagellar protein trafficking deeper into the Chlamydomonas cell body, a process presumably tightly coupled to IFT. This thesis project explored 1) an array of fluorescent proteins with various intensities and spectra, 2) filter sets to distinguish signals against background, and 3) a new strategy for tagging a protein of interest a string of fluorescent proteins. Through a combination of optimizations, a candidate new transit center and possible intracellular trafficking modalities have been identified. These pivotal findings will stimulate novel research questions that will address important questions in flagellar biology and will also make many obscured processes visible in the future.