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Jason P Affourtit

Publications and source records attributed to Jason P Affourtit.

2 recordsLinked to original sources

Transcriptional profile of mouse pre-granulosa and Sertoli cells isolated from early-differentiated fetal gonads.

Gonadal sex determining (GSD) genes that initiate fetal ovarian and testicular development and differentiation are expressed in the cells of the urogenital ridge that differentiate as somatic support cells (SSCs), i.e., granulosa cells of the ovary and Sertoli cells of the testis. To identify potential new mammalian GSD genes, we used the Mouse Genome 430v2.0 GeneChip to analyze gene expression differences between XX and XY SSCs cells isolated from the gonads of embryonic day (E) 13 C57BL/6J fetuses carrying an EGFP reporter transgene expressed specifically in SSCs. In addition, genome wide expression differences between XX and XY E13 whole gonads were examined. Analysis revealed that XX and XY E13 SSCs differentially express 647 transcripts (False Discovery Rate cutoff 1%), including transcripts not previously reported to exhibit a sexually dimorphic expression pattern in this unique cell population. Enrichment for genes controlling cell proliferation was noted in XY SSCs, whereas enrichment for genes controlling cell morphology and metabolic status was identified in XX SSCs. Among the newly identified differentially expressed transcripts are potential GSD genes involved in unexplained human sex reversal cases.

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Expression profiling using a hexamer-based universal microarray.

We describe a transcriptional analysis platform consisting of a universal micro-array system (UMAS) combined with an enzymatic manipulation step that is capable of generating expression profiles from any organism without requiring a priori species-specific knowledge of transcript sequences. The transcriptome is converted to cDNA and processed with restriction endonucleases to generate low-complexity pools (approximately 80-120) of equal length DNA fragments. The resulting material is amplified and detected with the UMAS system, comprising all possible 4,096 (4(6)) DNA hexamers. Ligation to the arrays yields thousands of 14-mer sequence tags. The compendium of signals from all pools in the array-of-universal arrays comprises a full-transcriptome expression profile. The technology was validated by analysis of the galactose response of Saccharomyces cerevisiae, and the resulting profiles showed excellent agreement with the literature and real-time PCR assays. The technology was also used to demonstrate expression profiling from a hybrid organism in a proof-of-concept experiment where a T-cell receptor gene was expressed in yeast.

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