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Published on: February 16, 2017
Microarray identification of novel downstream targets of FoxD4L1/D5, a critical component of the neural ectodermal
Bo Yan1, Karen M Neilson, Sally A Moody
1Department of Anatomy and Regenerative Biology, The George Washington University School of Medicine and Health Sciences, Washington, DC 20037, USA.
Abstract:
FoxD4L1/D5 is a forkhead transcription factor that functions as both a transcriptional activator and repressor. FoxD4L1/D5 acts upstream of several other neural transcription factors to maintain neural fate, regulate neural plate patterning, and delay the expression of neural differentiation factors. To identify a more complete list of downstream genes that participate in these earliest steps of neural ectodermal development, we carried out a microarray analysis comparing gene expression in control animal cap ectodermal explants (ACs), which will form epidermis, to that in FoxD4L1/D5-expressing ACs. Forty-four genes were tested for validation by RT-PCR of ACs and/or in situ hybridization assays in embryos; 86% of those genes up-regulated and 100% of those genes down-regulated in the microarray were altered accordingly in one of these independent assays. Eleven of these 44 genes are of unknown function, and we provide herein their developmental expression patterns to begin to reveal their roles in ectodermal development.
Insights
Forkhead transcription factor FoxD4L1/D5 plays a key role in early neural development. This study identified novel downstream genes regulated by FoxD4L1/D5, advancing our understanding of neural fate determination.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- FoxD4L1/D5 is a transcription factor crucial for maintaining neural fate and patterning.
- It acts upstream of other neural transcription factors, regulating early neural development.
- Understanding its downstream targets is essential for comprehending ectodermal development.
Purpose of the Study:
- To identify downstream genes regulated by FoxD4L1/D5 in early neural development.
- To characterize the expression patterns of novel genes involved in ectodermal development.
Main Methods:
- Microarray analysis comparing gene expression in control and FoxD4L1/D5-expressing animal cap explants.
- Validation of differentially expressed genes using RT-PCR and in situ hybridization.
- Analysis of developmental expression patterns for genes of unknown function.
Main Results:
- Microarray analysis revealed a set of genes regulated by FoxD4L1/D5.
- RT-PCR and in situ hybridization validated the microarray findings for 86% of upregulated and 100% of downregulated genes.
- Developmental expression patterns were determined for eleven previously uncharacterized genes.
Conclusions:
- FoxD4L1/D5 significantly influences the expression of numerous downstream genes during early ectodermal development.
- This study provides a foundational list of genes and their expression patterns, aiding future research into their specific roles.
- The identified genes and their patterns offer new insights into the molecular mechanisms governing neural fate and patterning.
