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| Status |
Public on Apr 03, 2018 |
| Title |
Comparative expression data between the epidermis and dermis of wild-type (Pparb/d^fl/fl) and fibroblast-selective knockout Pparb/d (FSPCre-Pparb/d^fl/fl) mice |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
The role of PPARβ/δ in maintaining skin homeostasis during skin injury or inflammation has been widely studied. However, the majority of these reports based their studies in PPARβ/δ found in keratinocytes, which is the major cell type of the skin epidermis. The skin is mainy made up of the epidermis and dermis, and skin homeostasis is tightly regulated by complex crosstalks between epidermis and dermis. The dermis is predominantly made up of fibroblasts, and the predominant PPAR subtype in dermal fibroblasts is PPARβ/δ. Knowledge in the role of fibroblasts PPARβ/δ in skin homeostasis is lacking.
To identify gene changes leading to phenotypical and biological functions alterations upon deletion of fibroblasts PPARβ/δ, we performed a comparative microarray gene expression analysis between Pparb/d^fl/fl and FSPCre-Pparb/d^fl/fl mice.
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| Overall design |
Pparb/d^fl/fl mice, having the exon 4 of their Pparb/d gene flanked by loxP sites, were mated with mice harbouring the Cre transgene under the control of the FSP promoter. FSPCre-Pparb/d^fl/+ progenies were then bred with Pparb/d^fl/fl mice to obtain FSPCre-Pparb/d^fl/fl mice and the wild-type control mice. To sustain this genotype, we bred FSPCre-Pparb/d^fl/fl progenies with Pparb/d^fl/fl mice. Mice were housed in a specific-pathogen free facility with a 12 h/12 h light/dark cycle. Food and water were provided ad libitum. Animal experiments were carried out in accordance to the guidelines of the University Institutional Animal Care and Use Committee (IACUC, ARF-SBS/NIE-A0216AZ), Singapore. Mice in the telogen phase of their hair cycle were anaesthetized and depilated on their dorsal back before being subjected to a biopsy punch (~4 mm). The skin biopsy was then treated with 3.8% ammonium thiocyanate (Sigma, USA) in 1X PBS for 30 min at 37 oC. Subsequently, the epidermis was mechanically separated from the dermis with a pair of forceps. The dermis was then curetted with a 30-gauge needle. RNA was extracted from the epidermis and dermis with Trizol according to the manufacturer’s protocol. Further sample processing of the RNA was carried out using Applause® WT-Amp ST System (NuGEN), and microarray experiments were performed on GeneChip® Mouse Gene 1.0 ST arrays according to the manufacturer’s instructions.
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| Contributor(s) |
Sng M |
| Citation(s) |
29619245 |
| |
| Submission date |
Jul 28, 2015 |
| Last update date |
May 30, 2019 |
| Contact name |
Ming Keat Sng |
| E-mail(s) |
mksng@ntu.edu.sg
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| Organization name |
Nanyang Technological University
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| Street address |
60 Nanyang Drive
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| City |
Singapore |
| ZIP/Postal code |
637551 |
| Country |
Singapore |
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| Platforms (1) |
| GPL6246 |
[MoGene-1_0-st] Affymetrix Mouse Gene 1.0 ST Array [transcript (gene) version] |
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| Samples (4)
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| GSM1833997 |
Fibroblast-specific Pparb/d knockout_Skin_Epidermis |
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| Relations |
| BioProject |
PRJNA291191 |
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