Regulation of high-affinity glutamate uptake activity in Bergmann glia cells by glutamate.
The effect of glutamate receptor activation on the high-affinity sodium-dependent glutamate transport expressed in chick Bergmann glia cells was examined. Pre-exposure to glutamate produced a time- and dose-dependent decrease in 3H-labeled D-aspartate uptake. This effect could not be reproduced by selective glutamate receptor agonists. Furthermore, it was insensitive to both ionotropic and metabotropic glutamate receptor antagonists. Replacement of extracellular sodium ions with choline in the preincubation media, abolished the reduction of the uptake. When the cells were pre-exposed to competitive transportable inhibitors of the transporter, such as D-aspartate, DL-threo-hydroxyaspartate (DL-THA), and aspartate-beta-hydroxamate (ABH), the glutamate effect was mimicked. From saturation experiments, it was found that the reduction on the uptake, after glutamate treatment, is related to an increase in K(m). Interestingly, the effect is blocked by staurosporine, a Ca(2+)/diacylglycerol-dependent protein kinase (PKC) inhibitor. The present findings suggest that glutamate regulates its transport in a non-receptor fashion, a phenomena that is most probably linked to changes induced by the translocation process of the substrate through the transporter.
González
M I
MI
Departamento de Genética y Biología Molecular, Cinvestav-IPN, Apartado Postal 14-740, DF 07000, México, Mexico.
Ortega
A
A
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D002525:Cerebellar Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002642:Chick Embryo; D018698:Glutamic Acid / Q000378:metabolism* / Q000494:pharmacology; D009457:Neuroglia / Q000166:cytology / Q000378:metabolism*; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism; D017470:Receptors, Glutamate / Q000187:drug effects / Q000378:metabolism; D014316:Tritium
D002352:Carrier Proteins; D017470:Receptors, Glutamate; D014316:Tritium; D001224:Aspartic Acid; D018698:Glutamic Acid; D011493:Protein Kinase C