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  • Title: Glucose deprivation causes posttranscriptional enhancement of brain capillary endothelial glucose transporter gene expression via GLUT1 mRNA stabilization.
    Author: Boado RJ, Pardridge WM.
    Journal: J Neurochem; 1993 Jun; 60(6):2290-6. PubMed ID: 8098356.
    Abstract:
    The absence of neuroglucopenia symptoms in chronic hypoglycemia may be due to up-regulation of the blood-brain barrier glucose transporter type 1 (GLUT1). Therefore, we investigated the effect of glucose deprivation on the abundance of the GLUT1 transcript in bovine brain capillary endothelial cells in tissue culture (ECL). Northern blot analysis performed under high stringency conditions with 4-5 micrograms of ECL poly(A)+ mRNA showed that glucose deprivation (5 mg% glucose) caused a 2.4 +/- 0.2-fold increase in the GLUT1/actin mRNA ratio versus control incubations (100 mg% glucose). This rise was dose and time dependent, and the maximum effect was observed 20-24 h after the hexose deprivation. Nuclear transcription run-on assay showed no changes in either the GLUT1 or actin gene transcription rate 24 h after glucose deprivation. To determine whether the increase in the abundance of the GLUT1 mRNA induced by glucose deprivation was due to increased stability of this transcript, the GLUT1 mRNA half-life was measured in ECL cells incubated with actinomycin D. The levels of the GLUT1 transcript continued to be augmented in glucose-deprived cells compared with controls 2 and 4 h after the transcription inhibitor was added to the media. Glucose deprivation induced a 78% increase in the t1/2 of the GLUT1 mRNA (from 3.6 to 6.4 h). Incubation of ECL cells with the protein synthesis inhibitor, cycloheximide, for 4 h partially reversed the effect of glucose deprivation on the abundance of the GLUT1 transcript. On the other hand, incubation with cycloheximide for 24 h completely blocked the effect of glucose deprivation on the GLUT1 transcript. Desensitization of cellular protein kinase C was performed by incubation of ECL cells with 1 microM phorbol ester for 24 h.(ABSTRACT TRUNCATED AT 250 WORDS)
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