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  • Title: Hypoxia induces de-stabilization of the LAT1 large neutral amino acid transporter mRNA in brain capillary endothelial cells.
    Author: Boado RJ, Li JY, Tsukamoto H, Pardridge WM.
    Journal: J Neurochem; 2003 May; 85(4):1037-42. PubMed ID: 12716435.
    Abstract:
    Blood-brain barrier (BBB) transport of large neutral amino acids is mediated by the large neutral amino acid transporter type 1 (LAT1 transporter). Although the gene encoding the Glut1 glucose transporter is up-regulated in hypoxia, the response of the LAT1 gene to hypoxia is not known. The present study investigates the changes in the LAT1 mRNA in cultured bovine brain capillary endothelial cells exposed to 1% O2 for 24-48 h. The LAT1 mRNA was initially down-regulated in hypoxia with reciprocal changes in the Glut1 mRNA. No changes in the 4F2hc mRNA in hypoxia were observed. Hypoxia caused an initial de-stabilization of the LAT1 mRNA, and the t1/2 of the LAT1 mRNA in control and hypoxic cells was 6.4 +/- 0.5 and 2.4 +/- 0.1 h, respectively. To further explore post-transcriptional regulation of LAT1 gene expression, the polysome and cytosol fractions of the control and hypoxic endothelial cells were isolated, and LAT1 mRNA binding proteins were detected by ultraviolet light cross-linking. Whereas the cytosol contained no LAT1 mRNA binding proteins, the cell polysome fraction expressed several LAT1 mRNA binding proteins, including principal 40-, 70- and 80-kDa proteins. These studies are consistent with post-transcriptional de-stabilization of the LAT1 large neutral amino acid transporter in hypoxia.
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