Epidermal growth factor and basic fibroblast growth factor protect dopaminergic neurons from glutamate toxicity in culture.
In this report we characterize the toxicity of the excitatory amino acid L-glutamate with respect to dopaminergic neurons cultured from embryonic rat mesencephalon. We also demonstrate that two growth factors, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), can protect these neurons from damage. Micromolar concentrations of L-glutamate, as well as agonists that specifically activate N-methyl-D-aspartate (NMDA) and non-NMDA receptors, are all toxic to dopamine neurons in a concentration-dependent manner, as reflected by decreases in high-affinity dopamine uptake and confirmed by decreases in numbers of tyrosine hydroxylase-immunoreactive neurons. Although the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione could attenuate the effects of quisqualate, treatment with this antagonist could not eliminate the effects of glutamate itself. Similarly, (+/-)-2-amino-5-phosphonopentanoic acid was effective against NMDA toxicity but could not protect cells from quisqualate toxicity. Thus, each type of receptor could mediate neurotoxicity independently of the other. The presence of EGF or bFGF in the culture medium conferred a relative resistance of dopaminergic neurons to glutamate and quisqualate neurotoxicity by increased glutamate transport. However, treatment of the cultures with L-trans-pyrrolidine-2,4-dicarboxylic acid, an inhibitor of glutamate transport, attenuated but did not eliminate the protective effects of both growth factors against glutamate toxicity. When cultures were incubated with conditioned medium from growth factor-treated cultures, neuroprotection was also achieved. These results suggest that both EGF and bFGF can protect neurons from neurotoxicity in culture by increasing the capacity of the culture for glutamate uptake as well as by the secretion of soluble factors into the medium.
Casper
D
D
Fishberg Research Center for Neurobiology, Mount Sinai School of Medicine, New York, New York, USA.
Blum
M
M
D000818:Animals; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004298:Dopamine / Q000378:metabolism*; D004622:Embryo, Mammalian; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D018698:Glutamic Acid / Q000633:toxicity*; D008648:Mesoderm / Q000187:drug effects; D009474:Neurons / Q000187:drug effects*; D016318:Quisqualic Acid / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D017077:Culture Media, Conditioned; D016194:Receptors, N-Methyl-D-Aspartate; D016222:Fibroblast Growth Factor 2; D018698:Glutamic Acid; D004815:Epidermal Growth Factor; D016318:Quisqualic Acid; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine