GDNF pre-treatment aggravates neuronal cell loss in oxygen-glucose deprived hippocampal slice cultures: a possible effect of glutamate transporter up-regulation.
Besides its neurotrophic and neuroprotective effects on dopaminergic neurons and spinal motoneurons, glial cell line-derived neurotrophic factor (GDNF) has potent neuroprotective effects in cerebral ischemia. The protective effect has so far been related to reduced activation of N-methyl-D-aspartate receptors (NMDAr). This study tested the effects of GDNF on glutamate transporter expression, with the hypothesis that modulation of glutamate transporter activity would affect the outcome of cerebral ischemia. Organotypic hippocampal slice cultures, derived from 1-week-old rats, were treated with 100 ng/ml GDNF for either 2 or 5 days, followed by Western blot analysis of NMDAr subunit 1 (NR1) and two glutamate transporter subtypes, GLAST and GLT-1. After 5-day exposure to GDNF, expression of GLAST and GLT-1 was up-regulated to 169 and 181% of control values, respectively, whereas NR1 was down-regulated to 64% of control. However, despite these changes that potentially would support neuronal resistance to excitotoxicity, the long-term treatment with GDNF was found to aggravate the neuronal damage induced by oxygen-glucose deprivation (OGD). The increased cell death, assessed by propidium iodide (PI) uptake, occurred not only among the most susceptible CA1 pyramidal cells, but also in CA3 and fascia dentata. Given that glutamate transporters are able to release glutamate by reversed action during energy failure, it is suggested that the observed increase in OGD-induced cell death in the GDNF-pretreated cultures was caused by the build-up of excitotoxic concentrations of extracellular glutamate released through the glutamate transporters, which were up-regulated by GDNF. Although the extent and consequences of glutamate release via reversal of GLAST and GLT-1 transporters seem to vary in different energy failure models, the present findings should be taken into account in clinical trials of GDNF.
Anatomy and Neurobiology, SDU-Odense University, Winsløwparken 21, DK-5000 Odense C, Denmark. cbonde@health.sdu.dk
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Bonde
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Sarup
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Schousboe
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Gegelashvili
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Noraberg
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Zimmer
D027322:Amino Acid Transport System X-AG / Q000378:metabolism*; D000818:Animals; D016923:Cell Death / Q000187:drug effects*; D027342:Excitatory Amino Acid Transporter 2 / Q000378:metabolism; D051100:Glial Cell Line-Derived Neurotrophic Factor; D005947:Glucose / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D066298:In Vitro Techniques; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D015854:Up-Regulation
D027322:Amino Acid Transport System X-AG; D027342:Excitatory Amino Acid Transporter 2; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; C409466:NR1 NMDA receptor; D009414:Nerve Growth Factors; D016194:Receptors, N-Methyl-D-Aspartate; D005947:Glucose; D010100:Oxygen