Janus Kinase (JAK) activity specifies the cell fates of the follicular epithelium during Drosophila oogenesis by establishing a gradient of JAK activity with highest levels at the A/P poles. Unpaired (Upd), a ligand for the pathway, is expressed and secreted exclusively from the polar cells potentially establishing the JAK activity gradient. This project proposed that Upd acts as a morphogen to directly establish the JAK activity gradient, specifying the fates of the follicular epithelium. The aims of this work were to investigate the extracellular distribution of Upd and, in addition, factors that may be involved. Furthermore, upd3, a gene encoding a protein with sequence similarity to Upd, is also co-expressed with upd in the polar cells. An additional aim of this project was to determine what role, if any, Upd3 plays in follicular development.
Immunostaining was used to reveal Upd distribution during oogenesis. The data revealed an Upd gradient on the apical membrane of the follicular epithelium. By virtue of the extracellular gradient, Upd fulfills the requirements necessary to be classified as a morphogen.
Some morphogens are dependent on heparan sulphate proteoglycans (HSPGs) for distribution. Using mitotic recombination to make mosaics, this work reveals that Dally, a glypican, is essential for the distribution of Upd and establishment of the JAK gradient during oogenesis. The data suggests Dally is involved with stability of extracellular Upd. Mosaic analysis of an additional HSPGs revealed that they are not essential for the Upd gradient or JAK activity during oogenesis.
upd3 mutant flies have small eyes and outstretched wings, a phenotype consistent reduced JAK activity. In upd3 mutant ovaries it is shown that there is a higher frequency of deteriorating egg chambers, a higher frequency of egg chamber fusions, and a decrease in border cells per egg chamber compared to wildtype controls; all of which support a reduction of JAK activity. Furthermore, ovarian phenotypes of upd3 get worse as the fly ages suggesting that upd3 is required over time. The data presented suggests that Upd3 does act to maintain JAK activity in the ovary as the fly ages.
Identifer | oai:union.ndltd.org:uky.edu/oai:uknowledge.uky.edu:gradschool_diss-1783 |
Date | 01 January 2009 |
Creators | Sexton, Travis |
Publisher | UKnowledge |
Source Sets | University of Kentucky |
Detected Language | English |
Type | text |
Format | application/pdf |
Source | University of Kentucky Doctoral Dissertations |
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