PUBLICATION
Depletion of the IKBKAP ortholog in zebrafish leads to hirschsprung disease-like phenotype
- Authors
- Cheng, W.W., Tang, C.S., Gui, H.S., So, M.T., Lui, V.C., Tam, P.K., Garcia-Barcelo, M.M.
- ID
- ZDB-PUB-150227-6
- Date
- 2015
- Source
- World journal of gastroenterology 21(7): 2040-6 (Journal)
- Registered Authors
- Keywords
- Enteric nervous system, Hirschsprung disease, IKBKAP, Morpholinos, Zebrafish
- MeSH Terms
-
- Carrier Proteins/genetics*
- Phenotype
- Genetic Predisposition to Disease
- Zebrafish Proteins/deficiency
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- Enteric Nervous System/embryology*
- Enteric Nervous System/metabolism
- Gene Knockdown Techniques
- Morpholinos/administration & dosage
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- Hirschsprung Disease/embryology
- Hirschsprung Disease/genetics*
- Hirschsprung Disease/metabolism
- Gene Expression Regulation, Developmental
- Immunohistochemistry
- Reverse Transcriptase Polymerase Chain Reaction
- Humans
- Homeodomain Proteins/genetics
- Homeodomain Proteins/metabolism
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Proto-Oncogene Proteins c-ret/genetics
- Proto-Oncogene Proteins c-ret/metabolism
- Animals
- Disease Models, Animal
- PubMed
- 25717236 Full text @ World J. Gastroenterol.
Citation
Cheng, W.W., Tang, C.S., Gui, H.S., So, M.T., Lui, V.C., Tam, P.K., Garcia-Barcelo, M.M. (2015) Depletion of the IKBKAP ortholog in zebrafish leads to hirschsprung disease-like phenotype. World journal of gastroenterology. 21(7):2040-6.
Abstract
Aim To investigate the role of IKBKAP (inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase complex-associated protein) in the development of enteric nervous system (ENS) and Hirschsprung disease (HSCR).
Methods In this study, we injected a morpholino that blocked the translation of ikbkap protein to 1-cell stage zebrafish embryos. The phenotype in the ENS was analysed by antibody staining of the pan-neuronal marker HuC/D followed by enteric neuron counting. The mean numbers of enteric neurons were compared between the morphant and the control. We also studied the expressions of ret and phox2bb, which are involved in ENS development, in the ikbkap morpholino injected embryos by quantitative reverse transcriptase polymerase chain reaction and compared them with the control.
Results We observed aganglionosis (χ (2), P < 0.01) and a reduced number of enteric neurons (38.8 ± 9.9 vs 50.2 ± 17.3, P < 0.05) in the zebrafish embryos injected with ikbkap translation-blocking morpholino (morphant) when compared with the control embryos. Specificity of the morpholino was confirmed by similar results obtained using a second non-overlapping morpholino that blocked the translation of ikbkap. We further studied the morphant by analysing the expression levels of genes involved in ENS development such as ret, phox2bb and sox10, and found that phox2bb, the ortholog of human PHOX2B, was significantly down-regulated (0.51 ± 0.15 vs 1.00 ± 0, P < 0.05). Although we also observed a reduction in the expression of ret, the difference was not significant.
Conclusion Loss of IKBKAP contributed to HSCR as demonstrated by functional analysis in zebrafish embryos.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping