Chronic treatment with sabeluzole protects cultured rat brain neurons from the neurotoxic effects of excitatory amino acids.
The neuroprotective properties of the cognitive enhancer sabeluzole were investigated in rat brain neuronal cultures derived from the hippocampal formation of 17-day-old rat embryos. Measurement of the neuronal cytoskeletal microtubule-associated protein, MAP2, was used to assess survival of neurons after exposure of neuronal cultures to glutamate. MAP2 was quantified in neuronal cell homogenates by means of an enzyme-linked immunosorbent assay (ELISA) using a mouse monoclonal MAP2 antibody, peroxidase-labeled goat anti-mouse Ig antiserum, and 2,2'-azido-di-[3-ethylbenz-thiazoline] sulphonate (ABTS) as substrate. Exposure of 7-day-old neuronal cultures to 1 mM glutamate for 16 hours led to a three-fold increase in released lactate dehydrogenase (LDH) and a 40% decrease in cellular MAP2 content. Acute treatment of neuronal cultures with 10 microM sabeluzole yielded a 40% drop in released LDH induced by glutamate. Cultures treated chronically with 0.1 microM sabeluzole on days 1 and 4 in culture showed, after 1 week in culture, a MAP2 content and total LDH activity that was not different from control cultures. A 16-hour exposure to 1 mM glutamate did not induce LDH release or changes in MAP2 levels in sabeluzole-treated cultures. A single treatment with 0.1 microM sabeluzole between day 1 to 5 induced a 70-80% drop in glutamate-induced released LDH in 7-day-old neuronal cultures. Full and partial neuronal protection after chronic sabeluzole treatment at 0.1 microM was also observed for neurotoxicity induced by 5 mM N-methyl-D-aspartate (NMDA) and 1 mM kainic acid or 30 microM veratridine, respectively. Within a series of compounds such as Ca++ and Na+ channel antagonists, glutamate receptor antagonists and various neurotransmitter receptor antagonists, sabeluzole, chronically given, were the most potent for inhibition of released LDH induced by 1 mM glutamate (IC50-value: 34 +/- 13 nM). In conclusion, chronic sabeluzole treatment protects cultured rat brain neurons from excitotoxic aggression.
Department of Biochemical Pharmacology, Janssen Research Foundation, Beerse, Belgium.
P J
PJ
Pauwels
H P
HP
Van Assouw
L
L
Peeters
M
M
Moeremans
J E
JE
Leysen
D000596:Amino Acids / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D000818:Animals; D001921:Brain / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002478:Cells, Cultured; D005971:Glutamates / Q000494:pharmacology; D018698:Glutamic Acid; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D010880:Piperidines / Q000494:pharmacology*; D051381:Rats; D013844:Thiazoles / Q000494:pharmacology*; D013997:Time Factors
D000596:Amino Acids; D005971:Glutamates; D008869:Microtubule-Associated Proteins; D010880:Piperidines; D013844:Thiazoles; D018698:Glutamic Acid; C055163:sabeluzole; D007770:L-Lactate Dehydrogenase