Kawata, M., Taniguchi, Y., Mori, D., Yano, F., Ohba, S., Chung, U. I., Shimogori, T., Mills, A. A., Tanaka, S., Saito, T. (2017) Different regulation of limb development by p63 transcript variants. PLoS One, 12 (3). e0174122. ISSN 1932-6203
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Abstract
The apical ectodermal ridge (AER), located at the distal end of each limb bud, is a key signaling center which controls outgrowth and patterning of the proximal-distal axis of the limb through secretion of various molecules. Fibroblast growth factors (FGFs), particularly Fgf8 and Fgf4, are representative molecules produced by AER cells, and essential to maintain the AER and cell proliferation in the underlying mesenchyme, meanwhile Jag2-Notch pathway negatively regulates the AER and limb development. p63, a transcription factor of the p53 family, is expressed in the AER and indispensable for limb formation. However, the underlying mechanisms and specific roles of p63 variants are unknown. Here, we quantified the expression of p63 variants in mouse limbs from embryonic day (E) 10.5 to E12.5, and found that DeltaNp63gamma was strongly expressed in limbs at all stages, while TAp63gamma expression was rapidly increased in the later stages. Fluorescence-activated cell sorting analysis of limb bud cells from reporter mouse embryos at E11.5 revealed that all variants were abundantly expressed in AER cells, and their expression was very low in mesenchymal cells. We then generated AER-specific p63 knockout mice by mating mice with a null and a flox allele of p63, and Msx2-Cre mice (Msx2-Cre;p63Delta/fl). Msx2-Cre;p63Delta/fl neonates showed limb malformation that was more obvious in distal elements. Expression of various AER-related genes was decreased in Msx2-Cre;p63Delta/fl limb buds and embryoid bodies formed by p63-knockdown induced pluripotent stem cells. Promoter analyses and chromatin immunoprecipitation assays demonstrated Fgf8 and Fgf4 as transcriptional targets of DeltaNp63gamma, and Jag2 as that of TAp63gamma. Furthermore, TAp63gamma overexpression exacerbated the phenotype of Msx2-Cre;p63Delta/fl mice. These data indicate that DeltaNp63 and TAp63 control limb development through transcriptional regulation of different target molecules with different roles in the AER. Our findings contribute to further understanding of the molecular network of limb development.
| Item Type: | Paper |
|---|---|
| Subjects: | organism description > animal > developmental stage > limb development bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > genes: types > p63 |
| CSHL Authors: | |
| Communities: | CSHL Cancer Center Program > Cancer Genetics CSHL labs > Mills lab CSHL Cancer Center Program > Gene Regulation and Inheritance Program |
| Depositing User: | Matt Covey |
| Date: | 2017 |
| Date Deposited: | 12 Apr 2017 15:56 |
| Last Modified: | 29 Oct 2020 19:08 |
| PMCID: | PMC5363923 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/34470 |
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