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Fatemeh Rajaii, Seth Blackshaw; Mouse orbital fat has a unique transcriptome compared to visceral, cutaneous, and brown fat depots, consistent with distinct developmental origins. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3000.
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© ARVO (1962-2015); The Authors (2016-present)
Thyroid associated orbitopathy (TAO), the most common orbital inflammatory disease, causes significant ocular morbidity due to orbital soft tissue fibrosis and adipogenesis. Mechanisms underlying orbital adipogenesis in TAO are not well-understood, however it is clear that orbital fat (OF) responds differently in TAO compared to other fat depots. We hypothesized that the distinct embryologic origin of OF is associated with unique properties in the adult and performed fate-mapping of neural crest in the mouse orbit to demonstrate that OF is neural crest-derived. We further performed comparative transcriptome analysis of murine OF and other fat depots to identify genes that are differentially expressed in OF.
Fate-mapping of craniofacial neural crest was performed using adult male Wnt1:Cre,Rosa26tdTomato mice. For transcriptome analysis, RNA was isolated from orbital, inguinal, epididymal, and brown fat of 9-week old CD-1 male mice. RNA sequencing performed on samples with RIN>7 using Illumina Nextseq500. Gene expression levels were compared across tissue samples. In situ hybridization was performed on adult CD-1 male orbits to validate gene expression.
tdTomato is expressed in OF in adult Wnt1:Cre,Rosa26tdTomato mice. Transcriptome analysis reveals that 306 genes are upregulated in OF compared to the other fat depots, and 29 genes are downregulated. Using in situ hybridization to validate gene expression demonstrates that selected genes identified in the screen are expressed in OF.
Murine orbital and periorbital fat are derived from neural crest and has a unique transcriptome compared to inguinal, epididymal, and brow fat. The distinct properties of OF may contribute to the unique response seen in OF in TAO.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
Anti-RFP staining of Wnt1:Cre Rosa26tdTomato mouse orbits demonstrates that tdTomato is expressed in OF (arrowhead in A) and periorbital fat (arrowhead in C).
A. Heatmap of OF-specific gene expression. B. In situ hybridization on 9 week old CD-1 males demonstrates expression of Fbxo40, IL20, Ky, Phex, Reep1, and Serpinb6d in OF.
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