The Journal of Neuroscience, October 12, 2005, ():
D-Serine Is the Dominant Endogenous Coagonist for NMDA Receptor Neurotoxicity in Organotypic Hippocampal Slices
J. Neurosci. Shleper et al.
25: 9413
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental figure1
Analysis of different D-amino acid oxidase preparations – A, SDS-Page analysis of D-amino acid oxidase (DAAOX) preparations. Each lane contained 3 μg protein. DAAOX from Boehringer Mannheim/Roche (DAAOX 1) was relatively pure, while Sigma (DAAOX 2) and Calzyme (DAAOX 3) preparations contained major contaminating protein bands. B and C, NMDA and D-serine oxidation by DAAOX preparations. The reaction was carried out in media containing 40 mM Tris-HCl (pH 8.5), 50 μM amplex red, 4 units/ml peroxidase and 15 μg/ml DAAOX 2 (B) or DAOOX 3 (C), either with 100 μM D-serine ( filled circle ) or NMDA ( open circle ). No NMDA oxidation was detected with DAAOX 1. The reaction was stopped by boiling for 3 minutes. Production of H2O2 resulting from D-serine or NMDA oxidation was monitored by the increase in amplex red fluorescence measured at 563 nm (excitation) and 587 nm (emission).
- supplemental material
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Supplemental figure 2
NMDA receptor 3A/B expression in organotypic hippocampal cultures- Each lane contains 20 μg protein from hippocampal formation obtained from postnatal day 4 rats before and after 10 days in culture. The western-blot analysis reveals NR3A/B band at about 125 kDa using a monoclonal anti-NR3A/B antibody at 5 μg/ml dilution (QED Bioscience, CA).
