The exocrine pancreas and intestine play essential roles in vertebrate physiology. For both organs cell renewal also has an important homeostatic role. In the intestine, cells of the villus epithelium are continuously replaced by the differentiated progeny of stem cell progenitors that reside within the crypts surrounding each villus. In the pancreas, multipotent progenitors reside within the ductular epithelium and believed to serve as a source of differentiated endocrine and acinar cells that can replace cells lost under specific experimental and pathological conditions. Identification of the genes that regulate cell lineage specification and differentiation of digestive organ stem cell progenitors is important to research related to common human diseases such as cancers of the pancreas and colon. Genetic analysis is an efficient means to identify genes that regulate developmental processes, particularly organogenesis. In this application we outline a strategy for the molecular characterization of lineage specification and cell differentiation in the developing exocrine pancreas and intestine of the zebrafish, a vertebrate model system uniquely suited to this purpose. The exocrine pancreas and intestine of zebrafish larvae function at five days post‑fertilization, a stage amenable to standard mutagenesis techniques. Further, progenitors and differentiated cells of both organs are easily visualized throughout development using organ specific markers.
The first Aim of this application is to perform a systematic mutagenesis screen for genes that regulate lineage specification and cell differentiation during exocrine pancreas and intestinal development. Using in situ and histochemical analysis we will identify genes that regulate growth and differentiation of the exocrine pancreas as well as development of the goblet and enteroendocrine cell lineages of the intestine. The feasibility of such as screen was proven in a recent pilot study.
The second Aim i s to identify molecular markers in the exocrine pancreas and intestine of larval zebrafjsh by generating a zebrafish pancreas and intestine EST database.
This aim will be performed in collaboration with members of the zebrafish genome community.
The third Aim i s to map the mutations and ESTs using meiotic and physical mapping techniques.
This aim will allow identification of candidate genes for the mutations recovered in Specific Aim 1, and provide a databank of targets for gene knockdown experiments.
The fourth aim of this application is to explore the function of novel genes identified in Specific Aim 2 using a morpholino based gene knockdown strategy.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK061142-05
Application #
6936052
Study Section
Special Emphasis Panel (ZDK1-GRB-4 (O1))
Program Officer
Karp, Robert W
Project Start
2001-09-30
Project End
2006-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
5
Fiscal Year
2005
Total Cost
$396,250
Indirect Cost
Name
University of Pennsylvania
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Gao, Nan; Davuluri, Gangarao; Gong, Weilong et al. (2011) The nuclear pore complex protein Elys is required for genome stability in mouse intestinal epithelial progenitor cells. Gastroenterology 140:1547-55.e10
Matthews, Randolph P; Eauclaire, Steven F; Mugnier, Monica et al. (2011) DNA hypomethylation causes bile duct defects in zebrafish and is a distinguishing feature of infantile biliary atresia. Hepatology 53:905-14
Matthews, Randolph P; Lorent, Kristin; Manoral-Mobias, Rafael et al. (2009) TNFalpha-dependent hepatic steatosis and liver degeneration caused by mutation of zebrafish S-adenosylhomocysteine hydrolase. Development 136:865-75
Davuluri, Gangarao; Gong, Weilong; Yusuff, Shamila et al. (2008) Mutation of the zebrafish nucleoporin elys sensitizes tissue progenitors to replication stress. PLoS Genet 4:e1000240
Matthews, Randolph P; Lorent, Kristin; Pack, Michael (2008) Transcription factor onecut3 regulates intrahepatic biliary development in zebrafish. Dev Dyn 237:124-31
Yee, Nelson S; Gong, Weilong; Huang, Ying et al. (2007) Mutation of RNA Pol III subunit rpc2/polr3b Leads to Deficiency of Subunit Rpc11 and disrupts zebrafish digestive development. PLoS Biol 5:e312
Wallace, Kenneth N; Akhter, Shafinaz; Smith, Erin M et al. (2005) Intestinal growth and differentiation in zebrafish. Mech Dev 122:157-73
Yee, Nelson S; Pack, Michael (2005) Zebrafish as a model for pancreatic cancer research. Methods Mol Med 103:273-98
Wallace, Kenneth N; Dolan, Amy C; Seiler, Christoph et al. (2005) Mutation of smooth muscle myosin causes epithelial invasion and cystic expansion of the zebrafish intestine. Dev Cell 8:717-26
Yee, Nelson S; Lorent, Kristin; Pack, Michael (2005) Exocrine pancreas development in zebrafish. Dev Biol 284:84-101

Showing the most recent 10 out of 12 publications