The liver performs several essential functions including the storage and filtration of blood, the secretion of bile, the synthesis and secretion of amino acids, and the conversions of sugars into glycogen. Despite these essential functions, little is known about the cellular and molecular mechanisms leading to the formation of the liver. We propose to use a forward genetic approach in zebrafish to investigate liver development. The gut tube ad its associated organs are not readily visible in the developing zebrafish embryo and thus have not been very accessible to morphological screens. Using a transgenic line which expresses GFP in the developing gut tube and its associated organs, the liver and pancreas, we plan to conduct a large-scale diploid screen for mutations that affect the formation of these organs. We will focus our future studies on mutations that affect the development of the liver. However, mutations that affect the formation of the pancreas and/or intestine will also be identified in our screen. We will set up a system to publish the phenotype of these mutations on the Web and make them available as soon as they are identified. In order to screen for mutations affecting the development of the liver, pancreas and intestine using this GFP transgenic line, we propose the following specific aims: 1) Characterize zebrafish liver development in detail. These studies will include further analysis of the GFP transgenic line as well as detailed examination of liver gene expression in wild-type and specific mutant backgrounds. 2) Screen for mutations that affect the formation of the liver, pancreas, and intestine. We will conduct a large- scale diploid screen using the GFP transgenic line. 3) Classify the liver mutations and begin to analyze several of them in detail. Complementation analysis, genetic mapping and molecular characterization will allow us to classify these mutations and select a few for further work. These molecular genetic studies of liver development will help provide the necessary context for further investigating the pathophysiology of malformations and malfunctions of the human liver, and may also facilitate differentiation-based strategies for treatment of a variety of diseased states.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK060322-05
Application #
6932497
Study Section
Special Emphasis Panel (ZRG1-BIOL-1 (02))
Program Officer
Karp, Robert W
Project Start
2001-06-01
Project End
2006-05-31
Budget Start
2005-06-01
Budget End
2006-05-31
Support Year
5
Fiscal Year
2005
Total Cost
$329,717
Indirect Cost
Name
University of California San Francisco
Department
Biochemistry
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
So, Juhoon; Khaliq, Mehwish; Evason, Kimberley et al. (2018) Wnt/?-catenin signaling controls intrahepatic biliary network formation in zebrafish by regulating notch activity. Hepatology 67:2352-2366
Choi, Tae-Young; Khaliq, Mehwish; Tsurusaki, Shinya et al. (2017) Bone morphogenetic protein signaling governs biliary-driven liver regeneration in zebrafish through tbx2b and id2a. Hepatology 66:1616-1630
Cox, Andrew G; Hwang, Katie L; Brown, Kristin K et al. (2016) Yap reprograms glutamine metabolism to increase nucleotide biosynthesis and enable liver growth. Nat Cell Biol 18:886-896
Gut, Philipp; Stainier, Didier Y R (2015) Whole-organism screening for modulators of fasting metabolism using transgenic zebrafish. Methods Mol Biol 1263:157-65
Evason, Kimberley J; Francisco, Macrina T; Juric, Vladislava et al. (2015) Identification of Chemical Inhibitors of ?-Catenin-Driven Liver Tumorigenesis in Zebrafish. PLoS Genet 11:e1005305
Markmiller, Sebastian; Cloonan, Nicole; Lardelli, Rea M et al. (2014) Minor class splicing shapes the zebrafish transcriptome during development. Proc Natl Acad Sci U S A 111:3062-7
Choi, Tae-Young; Ninov, Nikolay; Stainier, Didier Y R et al. (2014) Extensive conversion of hepatic biliary epithelial cells to hepatocytes after near total loss of hepatocytes in zebrafish. Gastroenterology 146:776-88
Hochgreb-Hagele, Tatiana; Yin, Chunyue; Koo, Daniel E S et al. (2013) Laminin *1a controls distinct steps during the establishment of digestive organ laterality. Development 140:2734-45
Nussbaum, Justin M; Liu, Liuhong J; Hasan, Syeda A et al. (2013) Homeostatic generation of reactive oxygen species protects the zebrafish liver from steatosis. Hepatology 58:1326-38
Gut, Philipp; Baeza-Raja, Bernat; Andersson, Olov et al. (2013) Whole-organism screening for gluconeogenesis identifies activators of fasting metabolism. Nat Chem Biol 9:97-104

Showing the most recent 10 out of 33 publications