Antisense knockdown of GLAST, a glial glutamate transporter, compromises retinal function.
OBJECTIVE
To elucidate the role of the glial glutamate transporter GLAST, in the regulation of retinal function.


METHODS
Antisense oligonucleotides to GLAST were injected intravitreally into the left eye of Wistar rats. Sense oligonucleotides (control) were injected into the right eye over a period of 3 days. Scotopic flash electroretinograms were recorded over a 20-day period. To assay whether the antisense oligonucleotides caused a reduction in the expression or the activity of GLAST, retinas were exposed to D-aspartate, a nonendogenous substrate of glutamate transporters. The retinas were immunolabeled with specific antibodies for D-aspartate. Retinal GLAST and glutamate distributions also were determined immunocytochemically.


RESULTS
Antisense oligonucleotides markedly suppressed the electroretinogram b-wave, whereas sense oligonucleotides had no significant effect. Significant changes in the electroretinogram were apparent 5 days after injection of antisense oligonucleotide and were sustained for at least 20 days. A marked reduction of D-aspartate uptake into Muller cells of retinas that had been exposed to the antisense oligonucleotides 5 days previously suggests a reduction of GLAST activity. The retinas, however, displayed no evidence of excitotoxic neuronal degeneration, and the distribution of glutamate was unaffected by antisense treatment.


CONCLUSIONS
The observed lack of neuronal degeneration suggests that reduced glutamate uptake into Muller cells does not cause excitotoxic tissue damage. A direct perturbation of glutamatergic signaling is more likely, because the rapid clearance of glutamate is necessary for light elicited signaling between photoreceptors and bipolar cells. This suggests that GLAST is essential for the maintenance of normal retinal transmission.
Barnett
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Vision, Touch, and Hearing Research Centre, The University of Queensland, Brisbane, Australia. n.barnett@vthrc.uq.edu.au
Pow
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D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000502:physiology*; D027322:Amino Acid Transport System X-AG; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001692:Biological Transport; D004596:Electroretinography / Q000187:drug effects; D005260:Female; D018698:Glutamic Acid / Q000378:metabolism*; D007124:Immunoenzyme Techniques; D009457:Neuroglia / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000187:drug effects* / Q000502:physiology*
D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D016376:Oligonucleotides, Antisense; D001224:Aspartic Acid; D018698:Glutamic Acid