Nine 5 RACE clones from male gonadal fat were isolated and identified as CHIR-258 Cyp19a1 transcripts. The nucleotide sequence of the 5 Inhibitors,Modulators,Libraries region of aromatase adipose specific transcript is shown in Figure 3B. The untranslated first exon in gonadal fat was 93 bp and was located 74763 bp upstream of the transla tional start site. The 5 UTRs of the 9 isolated clones were identical and were not similar to any previously known Cyp19a1 transcript sequence. 5 RACE clones from 3 other tissues were analyzed as controls. Five clones from the ovary had identical transcriptional start sites, and the sequences of the 5 UTRs Inhibitors,Modulators,Libraries of the ovarian transcripts were identical to ovarian Cyp19a1 cDNA described previously. Cyp19a1 transcripts from 8 5 RACE clones were identified as brain aromatase cDNA.

The transcription start site in male mice was at posi tion 382 of EMBL GenBank accession number D67045, which was 47 nucleotides upstream Inhibitors,Modulators,Libraries from that described previously. The transcription start site in female mice was at position 444 of EMBL GenBank accession number D67045, 15 nucleotides downstream from that described previously. The transcription start site of the testis specific transcript was located at position 1589 of EMBL accession number AJ437576, which was 18 nucle otides downstream from that described by Vanselow et al.The small discrepancy between our results and pre vious published Inhibitors,Modulators,Libraries data with respect to the presumed tran scription start sites of brain and testis specific Cyp19a1 mRNAs may be due to a range of possible initiation sites within a confined region or slightly different techniques used in various laboratories.

Expression of mouse Cyp19a1 adipose specific Inhibitors,Modulators,Libraries transcript in various tissues To determine the distribution of Tad, Tov, Ttes, and Tbr in male gonadal fat, ovary, testes, and hypothalamus, exon specific RT PCR analysis of RNAs was performed using 5 in 10 and 16 week old male UTR specific forward primers and reverse primers located in coding exon II. Tad and Tov were expressed in male gonadal fat. Those results were confirmed by real time RT PCR in 10 week old and 16 week old mouse gonadal fat. Tbr and Tov were found in the hypothalamus. Tov was the only Cyp19a1 transcript expressed in the ovary. All 4 transcripts were found in the testis, though Tov was the major transcript. Thus, we sug gested renaming the previously described Tov as gonadal specific transcript.

Male gonadal adipose specific promoter activity In order to characterize promoter activity of the newly identified male gonadal fat specific transcript, we gener ated 2 luciferase reporter constructs containing 343 1 and 1637 1 bp of the 5 sequence and assessed their activity in luciferase reporter assays. As compared with the empty control vector, luciferase nearly activity was increased by 1. 7 fold in cells transfected with the 343 1 construct, and decreased by 3.