PUBLICATION
On the traces of tcf12: Investigation of the gene expression pattern during development and cranial suture patterning in zebrafish (Danio rerio)
- Authors
- Blümel, R., Zink, M., Klopocki, E., Liedtke, D.
- ID
- ZDB-PUB-190613-11
- Date
- 2019
- Source
- PLoS One 14: e0218286 (Journal)
- Registered Authors
- Blümel, Rabea, Klopocki, Eva, Liedtke, Daniel
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Basic Helix-Loop-Helix Transcription Factors/genetics*
- Cranial Sutures/growth & development
- Craniosynostoses/genetics
- Embryo, Nonmammalian/physiology*
- Embryonic Development/genetics*
- Gene Expression Regulation, Developmental/genetics*
- Osteogenesis/physiology
- Promoter Regions, Genetic/genetics
- Transcription Factors/genetics
- Zebrafish/genetics*
- Zebrafish/growth & development*
- Zebrafish Proteins/genetics*
- PubMed
- 31188878 Full text @ PLoS One
Citation
Blümel, R., Zink, M., Klopocki, E., Liedtke, D. (2019) On the traces of tcf12: Investigation of the gene expression pattern during development and cranial suture patterning in zebrafish (Danio rerio). PLoS One. 14:e0218286.
Abstract
The transcription factor 12 (tcf12) is a basic Helix-Loop-Helix protein (bHLH) of the E-protein family, proven to play an important role in developmental processes like neurogenesis, mesoderm formation, and cranial vault development. In humans, mutations in TCF12 lead to craniosynostosis, a congenital birth disorder characterized by the premature fusion of one or several of the cranial sutures. Current research has been primarily focused on functional studies of TCF12, hence the cellular expression profile of this gene during embryonic development and early stages of ossification remains poorly understood. Here we present the establishment and detailed analysis of two transgenic tcf12:EGFP fluorescent zebrafish (Danio rerio) reporter lines. Using these transgenic lines, we analyzed the general spatiotemporal expression pattern of tcf12 during different developmental stages and put emphasis on skeletal development and cranial suture patterning. We identified robust tcf12 promoter-driven EGFP expression in the central nervous system (CNS), the heart, the pronephros, and the somites of zebrafish embryos. Additionally, expression was observed inside the muscles and bones of the viscerocranium in juvenile and adult fish. During cranial vault development, the transgenic fish show a high amount of tcf12 expressing cells at the growth fronts of the ossifying frontal and parietal bones and inside the emerging cranial sutures. Subsequently, we tested the transcriptional activity of three evolutionary conserved non-coding elements (CNEs) located in the tcf12 locus by transient transgenic assays and compared their in vivo activity to the expression pattern determined in the transgenic tcf12:EGFP lines. We could validate two of them as tcf12 enhancer elements driving specific gene expression in the CNS during embryogenesis. Our newly established transgenic lines enhance the understanding of tcf12 gene regulation and open up the possibilities for further functional investigation of these novel tcf12 enhancer elements in zebrafish.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping