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Published on: November 2, 2013
Application of differential display in the identification of androgen-regulated genes
Karen J Greenland1, Georgia Rekaris, Helen E MacLean
1Centre for Hormone Research, Murdoch Children's Research Institute, University of Melbourne, Royal Children's Hospital, Victoria, Australia.
Abstract:
Identification of androgen-regulated genes in neurons is an important step in understanding the mechanisms involved in androgen action. The aim of the current study was to identify androgen-responsive genes in the neural cells using the technique of differential display reverse transcription polymerase chain reaction (DDRT-PCR) on the human neuroblastoma cell line, SK-N-MC. Using this analysis, 18 putatively androgen-regulated cDNA species were identified, ranging in size from 280 to 800 bp. Of these, 14 were found to be negatively regulated and 4 positively regulated by androgens. Only 12 were successfully re-amplified, and of these, 8 were found to contain multiple species of cDNA fragments. When Northern analysis was conducted using the 21 different cDNA fragments as probes, only one was found to confirm the androgen regulation demonstrated via DDRT-PCR. While this putatively regulated gene remains to be fully characterized, future studies of may provide insights into the molecular mechanisms governing androgen action in neural cells.
