Unknown,Transcriptomics,Genomics,Proteomics

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BMP4 effect on gene expression program in human primary epidermal keratinocytes and dermal fibroblasts

ABSTRACT: BMP signalling is a potent regulator of skin morphogenesis, homeostasis and remodelling. However, molecular mechanisms underlying its involvement in regulating gene expression programs in keratinocytes and fibroblasts remain largely unknown. We analyzed the effect of BMP4 tratment on gene expression programs in human primary epidermal keratinocyte and dermal fibroblasts cultures. We identified specific changes in gene expression programs for each cell type. The primary human epidermal keratinocytes and dermal fibroblasts were treated with recombinant BMP4 or solvent control for 8 hours to reveal early and intermediate response genes. The RNA was isolated and used for micro-array analysis. Changes in gene expression programs were analyzed for each cell type and were compared between cell types.

ORGANISM(S): Homo sapiens

SUBMITTER: Michael Fessing

PROVIDER: E-GEOD-16111 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Hair follicle formation depends on reciprocal epidermal-dermal interactions and occurs during skin development, but not in adult life. This suggests that the properties of dermal fibroblasts change during postnatal development. To examine this, we used a PdgfraEGFP mouse line to isolate GFP-positive fibroblasts from neonatal skin, adult telogen and anagen skin and adult skin in which ectopic hair follicles had been induced (EF skin) by transgenic epidermal activation of beta-catenin. We also isolated epidermal cells from each mouse. The gene expression profile of EF epidermis was most similar to that of anagen epidermis, consistent with activation of beta-catenin signalling. In contrast, adult dermis with ectopic hair follicles more closely resembled neonatal dermis than adult telogen or anagen dermis. In particular, genes associated with mitosis were upregulated and extracellular matrix-associated genes were downregulated in neonatal and EF fibroblasts. We confirmed that sustained epidermal beta-catenin activation stimulated fibroblasts to proliferate to reach the high cell density of neonatal skin. In addition, the extracellular matrix was comprehensively remodelled, with mature collagen being replaced by collagen subtypes normally present only in developing skin. The changes in proliferation and extracellular matrix composition originated from a specific subpopulation of fibroblasts located beneath the sebaceous gland. Our results show that adult dermis is an unexpectedly plastic tissue that can be reprogrammed to acquire the molecular, cellular and structural characteristics of neonatal dermis in response to cues from the overlying epidermis. We have isolated the following populations of cells from mouse back skin by flow cytometry: 1A) GFP+ WT neonatal dermal fibroblasts, 1B) ItgA6+ WT neonatal epidermal keratinocytes, 2A) GFP+ WT telogen dermal fibroblasts, 2B) ItgA6+ WT telogen epidermal keratinocytes, 3A) GFP+ D2 transient activation (anagen) dermal fibroblasts, 3B) ItgA6+ D2 transient activation (anagen) epidermal keratinocytes, 4A) GFP+ D2 sustained activation (ectopic follicles) dermal fibroblasts, 4B) ItgA6+ D2 sustained activation (ectopic follicles) epidermal keratinocytes

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BMP4 effect on gene expression program in human primary epidermal keratinocytes and dermal fibroblasts

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