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Microgavage of Zebrafish Larvae
Published on: February 20, 2013
FACS-assisted microarray profiling implicates novel genes and pathways in zebrafish gastrointestinal tract
Carsten Stuckenholz1, Lili Lu, Prakash Thakur
1Department of Medicine, Division of Hematology/Oncology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
Gastroenterology
|July 1, 2009
Summary
Researchers profiled zebrafish gastrointestinal development to find key genes. This study identified novel genes and pathways crucial for organogenesis, offering potential therapeutic targets for gastrointestinal diseases and cancers.
Area of Science:
- Developmental Biology
- Genomics
- Molecular Biology
Background:
- The zebrafish (Danio rerio) serves as a valuable model for studying mammalian gastrointestinal development.
- Understanding the genetic regulation of gastrointestinal organogenesis is crucial for identifying disease mechanisms.
Purpose of the Study:
- To identify differentially regulated genes involved in zebrafish gastrointestinal organogenesis.
- To profile the transcriptome of the developing zebrafish gastrointestinal tract.
Main Methods:
- Utilized a transgenic zebrafish line expressing green fluorescent protein (GFP) in the intestine, liver, and pancreas.
- Employed fluorescence-activated cell sorting (FACS) to isolate GFP-expressing cells at four developmental time points.
- Performed microarray analysis and annotated the Zebrafish GeneChip with human orthologs.
Main Results:
- Transcriptional profiling revealed significant gene expression differences between GFP(+) and GFP(-) cells.
- Identified key pathways, including hepatic nuclear factor networks and the phosphatidylinositol 3 kinase (PI3K) pathway, relevant to mammalian gastrointestinal development and cancer.
- Discovered novel genes such as microRNAs (miR-217, miR-122), claudin c, fam136a, and a zebrafish tetraspanin.
- Identified a set of 32 genes potentially mediating the effects of chromosome arm 8q gain in human cancers.
Conclusions:
- Successfully combined FACS and microarray profiling to track organogenesis during development.
- Identified novel genes and pathways implicated in mammalian gastrointestinal development.
- These findings offer potential therapeutic targets for gastrointestinal diseases and cancers.
