The objective is to create a global gene expression atlas of the developing kidney. The central thesis is that a combination of laser capture microdissection and microarrays can be used to efficiently achieve this goal. Microarrays with essentially complete gene representation can be used to rapidly determine the expression levels of every gene in laser capture microdissected elements of the developing kidney. A single experiment, therefore, provides a comprehensive analysis of the gene expression status of one component, and a limited number of experiments examining each structure and substructure can create an atlas.
Specific aim 1 is to use this strategy to produce an atlas of the gene expression profiles of specific domains of the developing mouse kidney. The initial focus will be on the E15.5 kidney, which provides a single time point with multiple stages of nephrogenesis, but earlier time points will also be examined. A combination of structure, lectin staining, immunohistochemistry and transgenic GFP expression will be used to precisely identify specific components and lineages.
Specific aim 2 is to make transgenic mouse tools to promote both gene expression profiling and functional studies of kidney development. We propose to make a series of transgenic mice with specific promoters driving restricted expression of a Cre-GFP cassette. These mice will serve a dual purpose, to allow identification of additional discreet kidney components for Specific Aim 1 and to aid future domain specific gene knockout studies in the developing kidney.
Specific aim 3 is to perform bioinformatics analysis of the microarray data and to make results readily available to the research community. Microarrays produce large gene lists that need to be sifted. Analysis of the complex orchestrations of gene expression defined in Specific Aim 1 will provide deeper insight into the genetic basis of the development of the distinct parts of the nephron.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project--Cooperative Agreements (U01)
Project #
1U01DK070251-01
Application #
6877365
Study Section
Special Emphasis Panel (ZDK1-GRB-7 (O2))
Program Officer
Wilder, Elizabeth L
Project Start
2004-09-30
Project End
2009-07-31
Budget Start
2004-09-30
Budget End
2005-07-31
Support Year
1
Fiscal Year
2004
Total Cost
$359,953
Indirect Cost
Name
Cincinnati Children's Hospital Medical Center
Department
Type
DUNS #
071284913
City
Cincinnati
State
OH
Country
United States
Zip Code
45229
Magella, Bliss; Adam, Mike; Potter, Andrew S et al. (2018) Cross-platform single cell analysis of kidney development shows stromal cells express Gdnf. Dev Biol 434:36-47
Potter, S Steven; Brunskill, Eric W (2012) Laser capture. Methods Mol Biol 886:211-21
Brunskill, Eric W; Georgas, Kylie; Rumballe, Bree et al. (2011) Defining the molecular character of the developing and adult kidney podocyte. PLoS One 6:e24640
Potter, S Steven; Brunskill, Eric W; Patterson, Larry T (2011) Defining the genetic blueprint of kidney development. Pediatr Nephrol 26:1469-78
Brunskill, Eric W; Sequeira-Lopez, Maria Luisa S; Pentz, Ellen S et al. (2011) Genes that confer the identity of the renin cell. J Am Soc Nephrol 22:2213-25
Potter, S Steven; Brunskill, Eric W; Patterson, Larry T (2010) Microdissection of the gene expression codes driving nephrogenesis. Organogenesis 6:263-9
Abbas, Rime; Kombu, Rajan S; Dignam, David et al. (2010) Polyethylene glycol modified-albumin enhances the cold preservation properties of University of Wisconsin solution in rat liver and a hepatocyte cell line. J Surg Res 164:95-104
Brunskill, Eric W; Potter, S Steven (2010) Gene expression programs of mouse endothelial cells in kidney development and disease. PLoS One 5:e12034
Georgas, Kylie; Rumballe, Bree; Valerius, M Todd et al. (2009) Analysis of early nephron patterning reveals a role for distal RV proliferation in fusion to the ureteric tip via a cap mesenchyme-derived connecting segment. Dev Biol 332:273-86
Qu, Xiaoyan A; Gudivada, Ranga C; Jegga, Anil G et al. (2009) Inferring novel disease indications for known drugs by semantically linking drug action and disease mechanism relationships. BMC Bioinformatics 10 Suppl 5:S4

Showing the most recent 10 out of 11 publications