Related Experiment Video
Updated: Mar 30, 2026
Modeling Mucosal Candidiasis in Larval Zebrafish by Swimbladder Injection
Published on: November 27, 2014
Ibuprofen slows migration and inhibits bowel colonization by enteric nervous system precursors in zebrafish, chick
Ellen Merrick Schill1, Jonathan I Lake1, Olga A Tusheva1
1Department of Pediatrics, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110, USA.
Insights
Ibuprofen exposure during pregnancy may increase Hirschsprung disease (HSCR) risk by impairing enteric neural crest-derived cell (ENCDC) migration. This effect appears independent of COX enzymes and is specific to certain cell types and strains.
Area of Science:
- Developmental biology
- Neuroscience
- Pharmacology
Background:
- Hirschsprung disease (HSCR) is a congenital condition involving the absence of the enteric nervous system (ENS).
- HSCR has genetic links, but incomplete penetrance suggests environmental factors influence risk.
- Commonly used medications during pregnancy are potential environmental influences on HSCR.
Purpose of the Study:
- To investigate if medications used in early human pregnancy affect the risk of Hirschsprung disease.
- To determine the cellular and molecular mechanisms by which ibuprofen impacts enteric nervous system development.
Main Methods:
- Zebrafish were treated with common pregnancy medications to observe HSCR-like phenotypes.
- Fetal mouse gut slice cultures were used to assess the effects of ibuprofen on enteric neural crest-derived cell (ENCDC) migration.
- Inhibition of ROCK pathway and analysis of COX-deficient mice were employed to elucidate mechanisms.
- In vivo and in vitro colonization assays in zebrafish, mouse, and chick models were conducted.
Main Results:
- Ibuprofen exposure in zebrafish induced an HSCR-like absence of enteric neurons.
- Ibuprofen reduced migration, lamellipodia formation, and active RAC1/CDC42 in mouse ENCDC.
- ROCK inhibition partially reversed ibuprofen's effects on mouse ENCDC migration.
- Ibuprofen inhibited ENCDC colonization in mouse and chick models, with delayed onset and strain-specific effects.
- COX-independent mechanisms were suggested by normal colonization in Ptgs1/Ptgs2 deficient mice.
Conclusions:
- Ibuprofen inhibits ENCDC migration and bowel colonization in vitro and in vivo across multiple species.
- The effects of ibuprofen are cell-type and strain-specific, suggesting complex interactions.
- These findings raise concerns about ibuprofen potentially increasing Hirschsprung disease risk in genetically susceptible individuals.
Abstract:
Hirschsprung Disease (HSCR) is a potentially deadly birth defect characterized by the absence of the enteric nervous system (ENS) in distal bowel. Although HSCR has clear genetic causes, no HSCR-associated mutation is 100% penetrant, suggesting gene-gene and gene-environment interactions determine HSCR occurrence. To test the hypothesis that certain medicines might alter HSCR risk we treated zebrafish with medications commonly used during early human pregnancy and discovered that ibuprofen caused HSCR-like absence of enteric neurons in distal bowel. Using fetal CF-1 mouse gut slice cultures, we found that ibuprofen treated enteric neural crest-derived cells (ENCDC) had reduced migration, fewer lamellipodia and lower levels of active RAC1/CDC42. Additionally, inhibiting ROCK, a RHOA effector and known RAC1 antagonist, reversed ibuprofen effects on migrating mouse ENCDC in culture. Ibuprofen also inhibited colonization of Ret+/- mouse bowel by ENCDC in vivo and dramatically reduced bowel colonization by chick ENCDC in culture. Interestingly, ibuprofen did not affect ENCDC migration until after at least three hours of exposure. Furthermore, mice deficient in Ptgs1 (COX 1) and Ptgs2 (COX 2) had normal bowel colonization by ENCDC and normal ENCDC migration in vitro suggesting COX-independent effects. Consistent with selective and strain specific effects on ENCDC, ibuprofen did not affect migration of gut mesenchymal cells, NIH3T3, or WT C57BL/6 ENCDC, and did not affect dorsal root ganglion cell precursor migration in zebrafish. Thus, ibuprofen inhibits ENCDC migration in vitro and bowel colonization by ENCDC in vivo in zebrafish, mouse and chick, but there are cell type and strain specific responses. These data raise concern that ibuprofen may increase Hirschsprung disease risk in some genetically susceptible children.
