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Marlene F. Wade1 , Virendra B. Mahesh1 *, Ganapathy K. Bhat1 , Pedro L. Zamorano1 , Darrell W. Brann1 *
Department of Physiology and Endocrinology, Medical College of Georgia, Augusta, Georgia USA 1
The mRNA Differential Display Technique (mRNA D.D.) was utilized to identify genes which are differentially expressed in GT1-7 cells after phorbol 12-myristate, 13-acetate (PMA) treatment and which may be involved in the regulation of GnRH hormone secretion and mRNA synthesis. GT1-7 cells were grown to 80% confluency and subsequently exposed to 100nM PMA or vehicle for 16 hr. The RNAimage system (GenHunter Corp.) was utilized to conduct mRNA D.D. Four potentially differentially expressed cDNA candidates have been confirmed to be up-regulated by PMA treatment and have been sequenced and subjected to Basic Local Alignment Search Tool (BLAST) analysis. The cDNAs are significantly homologous to mRNA sequences encoding aldose reductase (AR), L10a ribosomal protein (L10a), and mouse brain potassium channel protein-1 (mBK1). One cDNA may encode a novel mRNA sequence. AR is an important enzyme in the polyol pathway which activates the pentose phosphate pathway, resulting in subsequent activation of the protein kinase C pathway which is known to regulate GnRH. L10a is a subunit of the 60S ribosomal protein. mBK1 is predicted to encode a protein significantly homologous to the Shaker protein, the A-component of a potassium channel. Further studies are ongoing to determine whether AR, L10a, and mBK1 are involved in GnRH regulation and to identify additional genes which are regulated by PMA in GT1-7 cells. This study was supported by a Research Grant (DWB) from the National Institute of Child Health and Human Development (RO1HD28964), NIH, US Public Health Service.
This abstract is being presented on Monday, August 2 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.