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Zebrafish as a model to analyze macromolecule absorption in intestinal enterocytes
Author(s) -
Cocchiaro Jordan L,
Navis Adam,
Bagnat Michel,
Rawls John F
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1148.23
Subject(s) - endocytosis , microbiology and biotechnology , endocytic cycle , vacuole , enterocyte , zebrafish , transcellular , macromolecule , pinocytosis , clathrin , paracellular transport , chemistry , biology , biophysics , biochemistry , small intestine , membrane , cell , cytoplasm , gene , permeability (electromagnetism)
Intestinal absorption and transcellular transport of macromolecules is important for neonatal passive immunity and development of oral tolerance to dietary antigens. The purpose of this study was to develop tools for live analysis of macromolecule absorption in the zebrafish intestine. Using a novel method for zebrafish gavage, we directly introduced macromolecules including immunoglobulin (IgG), dextran, and transferrin (Tnf) into the intestinal lumen of zebrafish larvae and performed live confocal microscopy to examine uptake and sorting within enterocytes. All macromolecules were initially absorbed into apical vesicles but subsequently partitioned within a larger supranuclear vacuole. In transgenic zebrafish expressing Lamp2‐RFP fusion, we found that Lamp2 was targeted to the vacuole membranes, and IgG and Tnf accumulated within these Lamp2‐ringed structures. Initial colocalization of macromolecules within the same apical vesicles suggested a non‐specific bulk endocytic mechanism; however, pretreatment of larvae with a panel of endocytosis inhibitors prior to gavage revealed that IgG and Tnf absorption were specifically blocked by chlorpromazine and phenylarsine oxide, inhibitors of clathrin‐mediated endocytosis. These results provide a zebrafish system to study the physiology and environmental regulation of macromolecule absorption and transport in the intestine. Research funded by NIDDK F32‐DK094592 to JLC and NIH RO1‐DK073695 awarded to JFR.