The $ beta$-tubulin gene family of the parasitic nematode, Brugia pahangi consists of three to five $ beta$-tubulin sequences. Two genomic clones containing $ beta$-tubulin sequences were isolated and characterized. The $ beta$1-tubulin gene spans 3.8 kb, is organized into 9 exons and expresses an mRNA of 1.7 kb which codes for a protein of 448 amino acids. A partial nucleotide sequence of the second clone confirmed the isolation of a distinct $ beta$-tubulin sequence, $ beta$2-tubulin. The $ beta$1-tubulin transcript is found in microfilariae and adult worms, whereas the $ beta$2-tubulin transcript is predominant in male adult worms but absent from microfilariae. Results of this study also indicate that the maturation of the $ beta$1-tubulin message involves the acquisition of the conserved nematode 22-nucleotide splice leader sequence. Antipeptide IgGs raised against the divergent carboxy-terminal region of $ beta$1-tubulin recognize the same $ beta$-tubulin isoform pattern as a phylum cross-reactive monoclonal antibody. This result suggests that the $ beta$1-tubulin is highly represented in B. pahangi adults and microfilariae.

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Cytokinesis is the physical division of one cell into two driven by an actomyosin contractile ring and positioned by signals from microtubules. This process is highly regulated spatially and temporally to ensure accurate division into two daughter cells. Here, I present work that builds upon our understanding of cytokinesis, focusing on the spatial requirements for protein function during cell division and midbody assembly. In Chapter 1, I present an introduction to cytokinesis and the cell and molecular mechanisms that govern the process. In Chapter 2, I present work I contributed to on the use of Upconverting nanoparticles for co-alignment of visible and infrared light on a light microscope. In Chapter 3, I present work developing a new microscopy technology called FLIRT (Fast Local Infrared Thermogenetics) that uses infrared light to inactivate fast-acting temperature sensitive protein function with subcellular precision and validate its use to study cytokinesis and cell fate signaling in the nematode Caenorhabditis elegans. In Chapter 4, I improve upon FLIRT technology by increasing its precision and demonstrate its use in studying the spatial regulation of key cytokinesis proteins including the actomyosin cytoskeleton in contractile ring constriction. The central spindle is an array of antiparallel overlapping microtubules that forms between the separating chromosomes in anaphase and is thought to serve as a signaling hub for cytokinesis. The central spindle is thought to become compacted during contractile ring constriction to form the dense midbody at the end of cell division. In Chapter 5, I investigate the requirements for central spindle microtubules in assembling midbodies in the C. elegans one-cell embryo. I present evidence that the CENP-F-like protein HCP-1 plays a primary role relative to its paralog HCP-2 in assembling the central spindle, and that the midbody can form independently of central spindle assembly. In Chapter 6, I discuss future directions for my work on both technology development and the mechanisms of cytokinesis. Through this work, I develop new technologies and hypotheses for how cytokinesis is spatially regulated within a cell, adding new complexity to our understanding of cell division.