? Numerous conditions in the neonate result in cholestasis, or impairment of bile flow. Understanding biliary development is key to understanding the mechanisms by which many of these diseases are caused, yet fairly little is known regarding biliary development at a molecular level. The zebrafish model has emerged as an excellent model to study development in general, and appears to share several features with mammals with respect to biliary development. We propose studies in which we can identify and characterize genetic pathways that regulate aspects of biliary development in the zebrafish. In the first specific aim, we expand our studies of hnf-6 in zebrafish biliary development. The transcription factor hnf-6 has been shown to be important in the remodeling of bile ducts during liver development in the mouse; we have found a similar role for hnf-6 in zebrafish. We propose a closer examination of the timing of hnf-6 expression during development. The second specific aim involves mapping and identification of the gene responsible for mda, a biliary development mutant identified in a mutagenesis screen utilizing a unique lipid reporter, PED-6. PED-6, a quenched fluorescent lipid, is swallowed by larval zebrafish, becomes unquenched in the intestine, and is eventually taken up by the liver and excreted into the bile, concentrating in the gallbladder. In the third aim, PED-6 will be used in ongoing efforts in this mutagenesis screen, looking for mutants that fail to concentrate PED-6 in the gallbladder, suggesting a possible defect in the anatomy of the bile ducts. ? The information that can be obtained through the studies outlined in this proposal should provide insight into molecular aspects of zebrafish biliary development. These studies in zebrafish may provide clues as to biliary development in mammals and to genetic contributions to cholestatic diseases in humans ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Clinical Investigator Award (CIA) (K08)
Project #
5K08DK068009-05
Application #
7483723
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Program Officer
Podskalny, Judith M,
Project Start
2004-09-15
Project End
2009-08-31
Budget Start
2008-09-01
Budget End
2009-08-31
Support Year
5
Fiscal Year
2008
Total Cost
$132,462
Indirect Cost
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
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Cui, Shuang; Capecci, Louis M; Matthews, Randolph P (2011) Disruption of planar cell polarity activity leads to developmental biliary defects. Dev Biol 351:229-41
Cui, Shuang; Erlichman, Jessi; Russo, Pierre et al. (2011) Intrahepatic biliary anomalies in a patient with Mowat-Wilson syndrome uncover a role for the zinc finger homeobox gene zfhx1b in vertebrate biliary development. J Pediatr Gastroenterol Nutr 52:339-44
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
Hand, Nicholas J; Master, Zankhana R; Eauclaire, Steven F et al. (2009) The microRNA-30 family is required for vertebrate hepatobiliary development. Gastroenterology 136:1081-90
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
Matthews, Randolph P; Lorent, Kristin; Pack, Michael (2008) Transcription factor onecut3 regulates intrahepatic biliary development in zebrafish. Dev Dyn 237:124-31
Matthews, Randolph P; Plumb-Rudewiez, Nicolas; Lorent, Kristin et al. (2005) Zebrafish vps33b, an ortholog of the gene responsible for human arthrogryposis-renal dysfunction-cholestasis syndrome, regulates biliary development downstream of the onecut transcription factor hnf6. Development 132:5295-306