This web page was produced as an assignment for Genetics 564, an undergraduate course at UW-Madison.
Autosomal dominant polycystic kidney disease (ADPKD) results in the formation of large numbers of kidney cysts impairing renal function [1]. Approximately 85% of ADPKD cases are caused by mutations in pkd1, which encodes a primary cilia protein [2]. When PKD1 function is disrupted, cyst proliferation occurs.
ADPKD is closely linked to development and aging. Gene ontology analysis of pkd1 indicates that it is important for a number of developmental processes in tissues throughout the body. Furthermore, evidence suggests that some cysts may even form within embryos [3]. The age at which cysts form is critical for understanding the pathogenesis of ADPKD, as cysts that form earlier have more time to grow, and thus, are larger. Although PKD1 has been linked to aging and development, its role during development is not currently clear.
The primary goal of this study is to determine the role of PKD1 during development. I hypothesize that pkd1 will have different expression patterns and that PKD1 will have different interaction partners at distinct developmental time points. The identification of differences in gene expression and protein interaction throughout development will yield insight in to the precise developmental control governing PKD1's function throughout the lifespan. A better understanding of differences in PKD1 function throughout the lifespan will then yield developmentally appropriate therapeutic targets.
This study will allow for a thorough developmental analysis of polycystic kidney disease using a bioinformatics approach. The long-term goal of this study is to identify differences in pkd1 expression or PKD1 function throughout development in order to yield potential therapeutic targets for a disease which has previously evaded proper treatment or cure. Ultimately, the predicted outcome of this study is better understanding of the development of disease, which will help to curb the devastating impact of a common hereditary disorder.
Specific Aim 1: To determine the expression pattern of PKD1 in developing tissues.
Approach: Microarray will be used to compare pkd1 expression in kidney, lung, skin, liver, and digestive tract in developing zebrafish embryos and in fully-formed adults.
Hypothesis: I expect that pkd1 will be expressed at high levels in the kidney and low levels in other tissues during development, since the effects of polycystic kidney disease are localized primarily to the kidneys.
Specific Aim 2: To determine whether knockdown of other kidney development genes can cause cystic kidneys.
Approach: Zebrafish embryos will be targeted with RNAi against genes associated with kidney development.
Hypothesis: I expect that cysts will develop upon knockdown of development-related PKD1 interaction partners. I also expect to identify interaction partners not previously known to interact with PKD1 or contribute to cyst development in kidneys.
Specific Aim 3: To determine if protein interactions with PKD1 are different in young and old zebrafish.
Approach: Tap-tag experiments to isolate proteins interacting with PKD1 in young and old zebrafish.
Hypothesis: I expect that PKD1's interaction partners will be different in young and old zebrafish and will reflect the function of PKD1 at different times during development.
ADPKD is closely linked to development and aging. Gene ontology analysis of pkd1 indicates that it is important for a number of developmental processes in tissues throughout the body. Furthermore, evidence suggests that some cysts may even form within embryos [3]. The age at which cysts form is critical for understanding the pathogenesis of ADPKD, as cysts that form earlier have more time to grow, and thus, are larger. Although PKD1 has been linked to aging and development, its role during development is not currently clear.
The primary goal of this study is to determine the role of PKD1 during development. I hypothesize that pkd1 will have different expression patterns and that PKD1 will have different interaction partners at distinct developmental time points. The identification of differences in gene expression and protein interaction throughout development will yield insight in to the precise developmental control governing PKD1's function throughout the lifespan. A better understanding of differences in PKD1 function throughout the lifespan will then yield developmentally appropriate therapeutic targets.
This study will allow for a thorough developmental analysis of polycystic kidney disease using a bioinformatics approach. The long-term goal of this study is to identify differences in pkd1 expression or PKD1 function throughout development in order to yield potential therapeutic targets for a disease which has previously evaded proper treatment or cure. Ultimately, the predicted outcome of this study is better understanding of the development of disease, which will help to curb the devastating impact of a common hereditary disorder.
Specific Aim 1: To determine the expression pattern of PKD1 in developing tissues.
Approach: Microarray will be used to compare pkd1 expression in kidney, lung, skin, liver, and digestive tract in developing zebrafish embryos and in fully-formed adults.
Hypothesis: I expect that pkd1 will be expressed at high levels in the kidney and low levels in other tissues during development, since the effects of polycystic kidney disease are localized primarily to the kidneys.
Specific Aim 2: To determine whether knockdown of other kidney development genes can cause cystic kidneys.
Approach: Zebrafish embryos will be targeted with RNAi against genes associated with kidney development.
Hypothesis: I expect that cysts will develop upon knockdown of development-related PKD1 interaction partners. I also expect to identify interaction partners not previously known to interact with PKD1 or contribute to cyst development in kidneys.
Specific Aim 3: To determine if protein interactions with PKD1 are different in young and old zebrafish.
Approach: Tap-tag experiments to isolate proteins interacting with PKD1 in young and old zebrafish.
Hypothesis: I expect that PKD1's interaction partners will be different in young and old zebrafish and will reflect the function of PKD1 at different times during development.
References[1] Gabow, P. (1993). Autosomal dominant polycystic kidney disease. The New England Journal of Medicine. 329(5) 332-342.
[2] Fedeles, S.V., Gallagher, A-R., Stefan Somlo. (2014) Polycystin-1: A master regulator of intersecting cystic pathways. Trends in Molecular Medicine. E-pub ahead of print. Doi: 10.1016/j.molmed.2014.01.004 [3] Reeders, S. T. et al. Prenatal diagnosis of autosomal dominant polycystic kidney disease with a DNA probe. Lancet 2, 6–8 (1986). [4] Banner: http://www.summitmedicalgroup.com/service/Nephrology/ |
Site created by: Elizabeth Roeske Last Updated: 2.4.2013 University of Wisconsin-Madison: Genetics 564 |