The long-term goals of this work are to understand cellular and molecular mechanisms of pancreas development during vertebrate embryogenesis. Over the past 2.5 years, we have conducted ascreen.for genes regulating pancreas development using a transgenic zebrafish line expressing GFP in the endoderm and endoderm-derived organs. We have also developed wholemount immunostaining techniques and utilized novel transgenic lines to characterize these mutants. Importantly, we carried out a detailed analysis of wild-type pancreas development in zebrafish (Field et al., 2003b), a study that has facilitated the use of the zebrafish system to identify and/or test genes implicated in this process. Specifically, we found that the zebrafish pancreas is formed by the fusion of two buds, a posterior one that contains only islet cells and an anterior one that gives rise to the exocrine cells and the pancreatic duct. The pancreatic duct in turn gives rise to new islet cells. This initial separation of endocrine and exocrine lineages should facilitate their investigation. We propose to continue our studies with the following specific aims. 1) Analyze the pancreas mutants identified and recovered from our screen. 2) Further characterize 6 mutants including detailed phenotypic analysis and isolation of the affected gene. 3) Analyze the role of the surrounding tissues in pancreas development and beta cell differentiation. We will analyze in detail the emergence of the initial islet cells (those that form the posterior bud) with the goals of identifying the signals that regulate this process. This information will be useful for designing cell culture protocols to differentiate stem cells into beta cells. We will also test the hypotheses that signals from the lateral plate mesoderm control the development of the anterior pancreatic bud, as well as the emergence of new islet cells from the pancreatic duct. 4) Continue to develop reagents and techniques to facilitate the use of the zebrafish as a rapid screening tool for genes regulating pancreas development and beta cell differentiation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
3R01DK075032-04S1
Application #
8034533
Study Section
Cellular Aspects of Diabetes and Obesity Study Section (CADO)
Program Officer
Sato, Sheryl M
Project Start
2010-04-06
Project End
2011-03-31
Budget Start
2010-04-06
Budget End
2011-03-31
Support Year
4
Fiscal Year
2010
Total Cost
$84,393
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
Lu, Jun-Xia; Bayro, Marvin J; Tycko, Robert (2016) Major Variations in HIV-1 Capsid Assembly Morphologies Involve Minor Variations in Molecular Structures of Structurally Ordered Protein Segments. J Biol Chem 291:13098-112
Tsuji, Naoki; Ninov, Nikolay; Delawary, Mina et al. (2014) Whole organism high content screening identifies stimulators of pancreatic beta-cell proliferation. PLoS One 9:e104112
Lancman, Joseph J; Zvenigorodsky, Natasha; Gates, Keith P et al. (2013) Specification of hepatopancreas progenitors in zebrafish by hnf1ba and wnt2bb. Development 140:2669-79
Ninov, Nikolay; Hesselson, Daniel; Gut, Philipp et al. (2013) Metabolic regulation of cellular plasticity in the pancreas. Curr Biol 23:1242-50
Andersson, Olov; Adams, Bruce A; Yoo, Daniel et al. (2012) Adenosine signaling promotes regeneration of pancreatic ? cells in vivo. Cell Metab 15:885-94
Delous, Marion; Yin, Chunyue; Shin, Donghun et al. (2012) Sox9b is a key regulator of pancreaticobiliary ductal system development. PLoS Genet 8:e1002754
Hesselson, Daniel; Anderson, Ryan M; Stainier, Didier Y R (2011) Suppression of Ptf1a activity induces acinar-to-endocrine conversion. Curr Biol 21:712-7
Tittle, Rachel K; Sze, Ryan; Ng, Anthony et al. (2011) Uhrf1 and Dnmt1 are required for development and maintenance of the zebrafish lens. Dev Biol 350:50-63
Kalousova, Anna; Mavropoulos, Anastasia; Adams, Bruce A et al. (2010) Dachshund homologues play a conserved role in islet cell development. Dev Biol 348:143-52
Yin, Chunyue; Kikuchi, Kazu; Hochgreb, Tatiana et al. (2010) Hand2 regulates extracellular matrix remodeling essential for gut-looping morphogenesis in zebrafish. Dev Cell 18:973-84

Showing the most recent 10 out of 20 publications