Kumar S, Sachar K, Huber J, Weingarten D P, de Vellis J
J Biol Chem. 1985 Nov 25;260(27):14743-7.
Utilizing a glycerol phosphate dehydrogenase cDNA clone from the rat glioma C6 cell line, we report that the glucocorticoid regulation of glycerol phosphate dehydrogenase gene expression in glial cells occurs at the transcriptional level. Furthermore, our transcription results in vitro, as well as Northern blot analysis, show that the short-chain fatty acid, sodium butyrate, totally blocks this hydrocortisone-induced transcription of glycerol phosphate dehydrogenase, demonstrating that the site of action of this fatty acid resides in the cell nucleus. The C6 glycerol phosphate dehydrogenase cDNA clone (pGPDH-1,1800 base pairs) used in these studies was selected from a C6 library generated from polysomal poly(A)+ RNA. pGPDH-1 hybridized to a 4.7-kilobase RNA from rat muscle and brain, mouse liver, and C6 cells; this mRNA is at least four times larger than the required coding sequence for glycerol phosphate dehydrogenase. Northern blot analysis of developing rat brain reveals a striking increase in glycerol phosphate dehydrogenase transcripts during the period most associated with peak myelination.
利用大鼠胶质瘤C6细胞系的磷酸甘油脱氢酶cDNA克隆，我们报道了神经胶质细胞中磷酸甘油脱氢酶基因表达的糖皮质激素调节发生在转录水平。此外，我们的体外转录结果以及Northern印迹分析表明，短链脂肪酸丁酸钠完全阻断了氢化可的松诱导的磷酸甘油脱氢酶转录，证明该脂肪酸的作用位点位于细胞核。这些研究中使用的C6磷酸甘油脱氢酶cDNA克隆（pGPDH-1，1800个碱基对）是从由多聚核糖体多聚（A）+RNA产生的C6文库中筛选出来的。pGPDH-1与大鼠肌肉和脑、小鼠肝脏及C6细胞中的4.7千碱基RNA杂交；这种mRNA比磷酸甘油脱氢酶所需的编码序列至少大四倍。对发育中的大鼠脑进行Northern印迹分析发现，在与髓鞘形成高峰期最相关的时期，磷酸甘油脱氢酶转录本显著增加。
