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• W2595243558 abstract "The bipotential nature of gonad progenitor cells provides an ideal model system to study the mechanisms of cell fate determination as one of two opposing fates are adopted: male or female. To better understand how supporting cells of the gonad initiate and stabilize this fate decision, we have begun investigating how chromatin is reorganized during sex determination. Within the nucleus, DNA is organized around nucleosomes which function to package DNA as well as regulate gene expression. Regions of the genome that are depleted for nucleosomes are indicative of various functional gene regulatory elements including promoters, enhancers, silencers and insulators. These regions are sensitive to digestion by the enzyme DNaseI, and DNaseI hypersensitivity assays have long been used to characterize and locate regulatory sequences. To understand how chromatin reorganizes as gonadal cells transition from a bipotential to differentiated state, we performed genome-wide DnaseI Hypersensitivity mapping (DNaseI-Seq). This assay allows us to delineate putative regulatory elements of critical sex determining genes and to investigate dynamic changes in nucleosome organization in an unbiased manner. Technical limitations of the technique, which require large numbers of purified cells, have restricted previous analyses to either cultured cell lines or adult primary cells. Here we present the first DNaseI-Seq analysis of a single embryonic cell lineage: differentiated XY preSertoli cells. We found that regions of nucleosome depletion can be identified in cells that have been frozen for extended periods of time, making purified embryonic cells amenable to DNaseI-Seq. Our data successfully identifies DNaseI hypersensitive sites (DHSs) at previously characterized regulatory elements and transcription factor binding sites. Using gene expression data from XX and XY gonadal cells before and after sex determination, we compared patterns of DHSs between gene regulatory loci that become either male- or female-specific during the sex determining process. We found that DHSs were enriched in regulatory domains surrounding genes that are specifically expressed or repressed in preSertoli cells compared to other XX or XY gonadal cell lineages. Therefore, DNaseI-Seq identifies functional regulatory elements important for sex determination and subsequent testis development. Future studies will examine dynamic changes in these patterns between XX and XY supporting cells of the gonad, comparing bipotential and differentiated stages. The causative gene for >50% of human disorders of sexual development is unknown and many are predicted to occur in distal regulatory sites. In addition to uncovering novel transcription factors based on motif screening of putative regulatory sequences, this analysis identifies discrete regions of the genome involved in sex determination and subsequent testis development that can inform better diagnostics for screening human patients with disorders of sexual development." @default.
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• W2595243558 date "2012-08-01" @default.
• W2595243558 modified "2023-09-24" @default.
• W2595243558 title "Using Chromatin Architecture to Identify Gene Regulatory Elements Required for Testis Development." @default.
• W2595243558 doi "https://doi.org/10.1093/biolreprod/87.s1.67" @default.
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