Investigating the Roll of Toll Receptors in Midline Development (2013)

Undergraduates: Nivedita Umasankar, Joseph Watson, Post-Doctoral Candidate

Faculty Advisor: Stephen Crews
Department: Biology

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The development of the nervous system is a highly complicated process that depends on complex cellular and chemical interactions. In the embryonic Drosophila midline, glia are one subset of cells that provide support to developing neurons, and understanding their migration is crucial to clarifying nervous system development. The emphasis of this project is to understand two pathways involved in glial migration: (1) to clarify the roles of two molecules, Spatzle 5 (spz5) and 18 wheeler (18w), within the NT-1 pathway, and (2) to generate antibodies for several toll-like receptors (TLRs) in order to determine their role in glial migration. Digoxygenin-labeled probes were generated for spz5 and 18w, and these probes were then used in fluorescent in-situ hybridizations of sim-Gal4 Uas-tau-GFP Drosophila embryos. The resulting images showed that spz5 was not significantly expressed in the midline, but that 18w was expressed in both the anterior and posterior midline glia, as well as several dividing neurons. For the second part of the experiment, antibodies were generated against four TLRs: Toll-6, Toll-7, Tollo, and 18W. Gene inserts of the TLRs were cloned into vectors, which were subsequently transformed into E. coli cells. Induction of these cells produced fusion proteins for each of the TLRs. These proteins were then injected into rabbits, and antibodies were successfully generated against three of the four TLRs, Toll-7, Tollo, and 18W.