Discovery of Transcription Factors Novel to Mouse Cerebellar Granule Cell Development Through Laser-Capture Microdissection
Laser-capture microdissection was used to isolate external germinal layer tissue from three developmental periods of mouse cerebellar development: embryonic days 13, 15, and 18. The cerebellar granule cell-enriched mRNA library was generated with next-generation sequencing using the Helicos technology. Our objective was to discover transcriptional regulators that could be important for the development of cerebellar granule cells—the most numerous neuron in the central nervous system. Through differential expression analysis, we have identified 82 differentially expressed transcription factors (TFs) from a total of 1311 differentially expressed genes. In addition, with TF-binding sequence analysis, we have identified 46 TF candidates that could be key regulators responsible for the variation in the granule cell transcriptome between developmental stages. Altogether, we identified 125 potential TFs (82 from differential expression analysis, 46 from motif analysis with 3 overlaps in the two sets). From this gene set, 37 TFs are considered novel due to the lack of previous knowledge about their roles in cerebellar development. The results from transcriptome-wide analyses were validated with existing online databases, qRT-PCR, and in situ hybridization. This study provides an initial insight into the TFs of cerebellar granule cells that might be important for development and provide valuable information for further functional studies on these transcriptional regulators.
KeywordsCerebellum Development Transcription Regulation Genetics LCM
We thank J. Yeung, J. Cairns, S. Tremblay, A. Poon, and J. Wilking for the support and suggestions on experimental design and manuscript preparation. We thank F. Lucero Villegas for animal management. We thank B. Lin, M. Larouche, D. Rains, and J. Boyle for technical support.
We thank National Institutes of Health, Natural Sciences and Engineering Research Council of Canada, NeuroDevNet, FANTOM OMICS Group, and University of British Columbia for funding.
Compliance with Ethical Standards
All experimentation with animals was under an approved Canadian Council on Animal Care research protocol (A12-0190).
Conflict of Interest
The authors declare that they have no conflict of interest.
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