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FAC U LTY O F H EALTH AN D M E D I CAL S C I E N C E S
UN IVE R S ITY O F C OPE NHAGE N
An oxazolone zebrafish model for inflammatory bowel disease (IBD):
preliminary results
Simon Haarder1, 3, Per Kania1, Poul Hyttel2, Thomas Lindebo Holm3 and Kurt Buchmann1
1 Department
of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark
2 Department
of Basic Animal and Veterinary Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Denmark
3 Department
of Immunopharmacology, Novo Nordisk A/S, Denmark
BACKGROUND AND AIM
The zebrafish (Danio rerio) is an popular animal model, increasingly used to investigate
basal pathogenesis of a variety of human diseases. Here, we use the zebrafish as a model
for inflammatory bowel disease (IBD), a chronic condition associated with the
gastrointestinal system.
Like other vertebrates, fish are equipped with a mucosal immune system in the skin (SALT). It
has recently been demonstrated that the immune response seen in fish skin in many aspects
resembles immune reactions in the fish gut. Hence, we wanted to clarify whether goblet
cells found in the epidermis (the outer part of the fish scale) were affected in the same way
as intestinal goblet cells are in zebrafish IBD models (depletion). If so, this parameter could
be included as a part of the overall disease model evaluation.
outer
inner
Scale dyed with Alcian Blue
OXAZOLONE CHALLENGE
• Wild-type zebrafish were
anaesthetized with MS-222
(200 mg/L) and weighed.
Control
• Fish were then challenged
with oxazolone (0.2, 0.6%,
1.0%, 5 fish per group, in 50%
ethanol (solvent)).
Day 0
Oxazolone
• Control fish were given 50%
ethanol.
• Fish were euthanized 3 days
post challenge (overdose MS222, 500 mg/L) and weighed.
Day 0
GOBLET CELL COUNTS
3 dpi
Control
Oxazolone
HISTOLOGY
0.2 %
No.
Goblet
cells *
3 dpi
Oxa
Control
216
254
0.6 %
Oxa
Control
1.0 %
Oxa
Kontrol
160
129
259
235
Control
Oxazolone
• Mid-intestine sections stained with Alcian Blue
• Goblet cells depletion seen in both groups
* Pooled from 5 different scales, Alcian blue staining
CONCLUSIONS
• Results indicate that goblet cells from fish scales are affected by oxazolone. Future zebrafish IBD models should include
this parameter in association with the traditional intestinal goblet cells counts .
• Histology data was inconclusive. Currently, alternative approaches are being investigated (e.g. whole-fish sectioning).
• Other methods are also performed on fish tissue: optimal projection tomography (OPT) and qPCR.
References:
- Brugmann et al. (2009): Oxazolone-Induced Enterocolitis in Zebrafish Depends on the Composition of the Intestinal Microbiota. Gastroenterology, 137, 1757-1767 pp.
- Xu et al. (2013): Teleost skin, an ancient mucosal surface that elicits gut-like immune responses. PNAS, 110 (32), 13097-13102 pp.