Regular articleSuramin inhibits β-bungarotoxin-induced activation of N-methyl-d-aspartate receptors and cytotoxicity in primary neurons
Section snippets
Materials
β-Bungarotoxin was isolated from the venom of the Bungarus multicinctus by the method described by Lee et al. (1972). The homogeneity of the purified toxins as revealed in a single band was verified by disc gel electrophoresis (David, 1964). This toxin was prepared in 1 mg/ml and stored at −70°C. Poly-l-lysine, trypsin, soybean trypsin inhibitor, cytosine arabino-side, DNase, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT), luciferin-luciferase, diltiazem, and carbonyl cyanide m
Effect of β-BuTX exposure on survival rate of cultured CGNs
The isolated CGNs were cultured for 7 days in vitro (DIV) to obtain maturation with a rich network of fasiculate fibers. We used seven DIV neurons throughout the whole experiment. The toxic effects of β-BuTX on survival rate of CGNs were concentration and time dependent (Fig. 1). After 24 h incubation, concentrations as low as 3 ng/ml of β-BuTX induced cytotoxicity. The EC50 estimated was 3 ng/ml (equivalent to 144 pM, Fig. 1A). However, exposure of astrocytes to 1 μg/ml β-BuTX did not result
Discussion
This study shows that β-BuTX is a potent toxicant to cultured CGNs. It was estimated that the toxic concentration of β-BuTX on cultured CGNs is nontoxic to mice in vivo in animals (Lin-Shiau and Lin, 1999). The selectivity of β-BuTX cytotoxicity to CGNs demonstrated that the cultured rat pheochromocytoma cell (PC12) and the human neuroblastoma cell (IMR32) were insensitive to β-BuTX (1 μg/ml, unpublished data). In this paper, we attempted to elucidate the possible molecular mechanism involved
Acknowledgments
This investigation was supported by a research grant (NSC90-2320-B-002-082) from the National Science Council, Taipei, Taiwan.
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