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Differential gene expression profile in skin of Hoxc13 gene-targeted mice

ABSTRACT: Hoxc13 has been shown to be essential for proper hair shaft differentiation as Hoxc13 gene-targeted (Hoxc13tm1Mrc) mice completely lack external hair. Because of the phenotypic parallels to the Foxn1nu (nude) mutant mice, we tested whether Hoxc13 and Foxn1 act in a common pathway of hair follicle (HF) differentiation. We show that the alopecia exhibited by both the Hoxc13tm1Mrc and Foxn1nu mice is due to strikingly similar defects in hair shaft differentiation and that both mutants suffer from a severe nail dystrophy. These similarities are consistent with the overlap of Hoxc13 and Foxn1 expression patterns in the HF and nail matrix. DNA microarray analysis of scapular skin from Hoxc13tm1Mrc mutant (mut) compared to wild type (wt) mice identified Foxn1 as significantly down-regulated along with numerous hair keratin genes. This Foxn1 down-regulation apparently reflects the loss of direct transcriptional control by HOXC13 as indicated by our results obtained through co-tranfection and chromatin immunoprecipitation (ChIP) assays.

ORGANISM(S): Mus musculus

PROVIDER: GSE23759 | GEO | 2010/08/25

SECONDARY ACCESSION(S): PRJNA130839

REPOSITORIES: GEO

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Differential gene expression profile in skin of Hoxc13 gene-targeted mice

Project description:Hoxc13 has been shown to be essential for proper hair shaft differentiation as Hoxc13 gene-targeted (Hoxc13tm1Mrc) mice completely lack external hair. Because of the phenotypic parallels to the Foxn1nu (nude) mutant mice, we tested whether Hoxc13 and Foxn1 act in a common pathway of hair follicle (HF) differentiation. We show that the alopecia exhibited by both the Hoxc13tm1Mrc and Foxn1nu mice is due to strikingly similar defects in hair shaft differentiation and that both mutants suffer from a severe nail dystrophy. These similarities are consistent with the overlap of Hoxc13 and Foxn1 expression patterns in the HF and nail matrix. DNA microarray analysis of scapular skin from Hoxc13tm1Mrc mutant (mut) compared to wild type (wt) mice identified Foxn1 as significantly down-regulated along with numerous hair keratin genes. This Foxn1 down-regulation apparently reflects the loss of direct transcriptional control by HOXC13 as indicated by our results obtained through co-tranfection and chromatin immunoprecipitation (ChIP) assays. Total number of samples was 6. The experiment was perfomed in triplicate involving 3 indendent wild type skin samples (sample 1, 2, and 3) and 3 independent mutant skin samples (sample 4, 5, and 6).

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Project description:rationale: comparison of gene expression profiles in wildtype and Foxn1::dnFGFR2-IIIb transgenic hair follicles; identification of targets that mediate the effects seen in transgenic hair follicles results: as already suggested by the phenotype, the molecular abnormalities appear to be restricted to the hair shaft medulla; Igfbp5 is an important mediator of transgene-dependent effects Keywords: ordered

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Transcription profiling of skin from wild type and Hoxc13 transgenic mice to investigate their propensity to develop severe hair growth defects and alopecia

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Bald thigh syndrome in sighthounds

Project description:Bald thigh syndrome is a common hair loss disorder in sighthounds. The overall goal of our study was to identify the cause of bald thigh syndrome and the pathological changes associated with it. We approached this aim by comparing skin biopsies and hair shafts of affected and control dogs microscopically as well as by applying high-throughput technologies such as genomics, transcriptomics and proteomics. While the histology is rather unspecific in most cases, trichogram analysis and scanning electron microscopy revealed severe structural abnormalities in hair shafts of affected dogs. This finding is supported by the results of the transcriptomic and proteomic profiling of skin biopsies and hair shafts, respectively, where genes and proteins important for differentiation of the inner root sheath and the assembly of a proper hair shaft were downregulated. Transcriptome profiling of skin biopsies revealed a downregulation of genes encoding 23 hair shaft keratins and 51 keratin associated proteins, as well as desmosomal cadherins and several actors of the BMP signaling pathway, which is important for hair shaft differentiation. To identify differentially expressed proteins in structural abnormal hair shafts, we performed nLC-MS/MS- based proteomic analysis of fractured hair shafts of four dogs with bald thigh syndrome (Greyhounds, n=3, Whippet, n=1) and intact telogen hair shafts of four control dogs (Greyhounds, n=3, Whippet= n=1) plucked on the thighs. Decreased expression of keratin 71 and desmocollin 2 on the mRNA level in skin biopsies corresponded with a reduced protein expression in the hair shafts of affected dogs.

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Project description:Quiescent hair follicle (HF) bulge stem cells (SCs) differentiate to early progenitor (EP) hair germ (HG) cells, which divide to produce transit-amplifying (TA) matrix cells. EPs can revert to SCs upon injury, but whether this de-differentiation occurs in normal HF homeostasis (hair cycle), and the mechanisms regulating both differentiation and de-differentiation are unclear. Here we use lineage tracing, gain of function, transcriptional profiling, and functional assays to examine the role of observed endogenous Runx1 level changes in the hair cycle. We find that forced Runx1 expression implements hair degeneration (catagen) and simultaneously promotes changes in the quiescent bulge SC transcriptome towards a cell-state resembling the EP HG fate. This cell-state transition is functionally reversible. We propose that SC differentiation and de-differentiation are likely to occur during normal HF degeneration and niche restructuring in response to changes in endogenous Runx1 levels associated with SC location with respect to the niche. Freshly isolated skin cells were FACS sorted based on K15-GFP+/a6-integrin+/CD34- as hair germ and K15-GFP+/a6-integrin+/CD34+ bulge cells. Duplicate samlpes from mice at PD20 that showed telogen morphology throughout skin were used for RNA prepartion and Affymetrix analysis. Wild and transgenic samples after 1-day of doxycycline treatement were compared.

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