This web page was produced as an assignment for Genetics 564, an undergraduate course at UW-Madison
Introduction
Gene ontology analysis of terms associated with PKD1 implicated many developmental-related processes in the development of polycystic kidney disease. The development of polycystic kidney disease proceeds throughout the lifetime, although the role of PKD1 early in development is not currently clear.
My primary goal for future research would be to analyze the connection between PKD1 and early development. I hypothesize that there are significant differences in PKD1 expression levels and polycystin-1 interaction partners at different developmental timepoints, and that the role of PKD1 thus evolves throughout the lifespan.
My primary goal for future research would be to analyze the connection between PKD1 and early development. I hypothesize that there are significant differences in PKD1 expression levels and polycystin-1 interaction partners at different developmental timepoints, and that the role of PKD1 thus evolves throughout the lifespan.
Expression analysis throughout development
My first goal is to determine the expression level of PKD1 in kidneys relative to other tissues during early and late development in zebrafish using a microarray analysis.
I expect that PKD1 will be expressed in high levels in the kidney and low levels in other tissues during development, primarily because the effects of polycystic kidney disease are localized to the kidneys.
This result will be significant because it will allow me to determine how important PKD1 expression is in other tissues during development.
I expect that PKD1 will be expressed in high levels in the kidney and low levels in other tissues during development, primarily because the effects of polycystic kidney disease are localized to the kidneys.
This result will be significant because it will allow me to determine how important PKD1 expression is in other tissues during development.
Screen for development-related proteins mediating cyst formation
My second goal is to determine the role of kidney-development genes in cyst formation. I plan to use an RNAi screen. I will add RNAs designed to knock down kidney-development genes to 96-well plates along with developing zebrafish embryos. I will then screen for the development of cystic kidneys in the resulting zebrafish.
I expect that cystic kidneys will form upon knockdown of several developmentally-related PKD1 interaction partners. Previous research has shown that cysts will form upon knockdown of PKD2, so I expect to see this result specifically.
This result will be important because it will identify polycystin-1 mediators involved in early development and distinct from later development.
I expect that cystic kidneys will form upon knockdown of several developmentally-related PKD1 interaction partners. Previous research has shown that cysts will form upon knockdown of PKD2, so I expect to see this result specifically.
This result will be important because it will identify polycystin-1 mediators involved in early development and distinct from later development.
Determine polycystin-1 interaction partners
My third goal is to determine polycystin-1 interaction partners specific to early and late development using a Tap-tag experimental approach.
I expect that polycystin-1 interaction partners will differ between young and old zebrafish.
This result will be significant because it will help to describe whether polycystin-1 acts differently early and late in development. This might be especially interesting since polycystin-1 is involved in signaling -- different interaction partners early and late in development might affect different signaling pathways early and late in development.
I expect that polycystin-1 interaction partners will differ between young and old zebrafish.
This result will be significant because it will help to describe whether polycystin-1 acts differently early and late in development. This might be especially interesting since polycystin-1 is involved in signaling -- different interaction partners early and late in development might affect different signaling pathways early and late in development.
Future Directions
My analysis will be limited to wild-type zebrafish embryos because my primary goal is to determine the role of PKD1 and polycystin-1 in normal developing animals. In the future, I could apply the results of these analyses to PKD1-mutant zebrafish backgrounds. In PKD1 mutant zebrafish, for example, expression in the kidneys during development might more closely mirror the expression of other tissues. Comparing the results of the wild-type zebrafish analysis and results in a PKD1-mutant background would help to confirm the results of this study and provide areas of future research.
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References
[1] cystic zebrafish kidney photo: http://: http://www.mayo.edu/research/labs/kidney-development-cyst-formation/overview [2] http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0035014 [3]Tag-trap photo: http://en.wikipedia.org/wiki/Tandem_affinity_purification |
Site created by: Elizabeth Roeske
Last updated: 2.4.2014 University of Wisconsin-Madison: Genetics 564 |