Method for measuring neurotoxicity of aggregating polypeptides with the MTT assay on differentiated neuroblastoma cells.
Reliable in vitro assays are essential for study of the effects of neurotoxic compounds such as beta-amyloid peptides (Abeta). The MTT assay has been used in cultures of different cells, e.g. SH-SY5Y neuroblastoma cells, for the quantitative measurement of Abeta toxicity. In our laboratory differentiated SH-SY5Y cells were used in the MTT assay. Cell differentiation with 10 microM all-trans-retinoic acid resulted in a constant cell number. The cells possess highly developed neurites and exhibit high sensitivity against Abeta. Owing to the constant cell number in differentiated SH-SY5Y cultures the decrease of the redox activity is directly proportional to the neurotoxicity of the substances, no correction is needed. The results of the MTT assay of Abeta peptides on differentiated SH-SY5Y cells displayed a good correlation also with the in vivo results. The present experiments reveal an effective assay for the study of potentially neurotoxic compounds.
Datki
Zsolt
Z
Department of Medical Chemistry, University of Szeged, Dóm tér 8, H-6720 Szeged, Hungary. datkiz@yahoo.com
Juhász
Anna
A
Gálfi
Márta
M
Soós
Katalin
K
Papp
Rita
R
Zádori
Dénes
D
Penke
Botond
B
D000682:Amyloid / Q000633:toxicity; D016229:Amyloid beta-Peptides / Q000633:toxicity*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D004396:Coloring Agents; D003584:Cytological Techniques; D006801:Humans; D058228:Islet Amyloid Polypeptide; D009447:Neuroblastoma; D010446:Peptide Fragments / Q000633:toxicity*; D011328:Prions / Q000633:toxicity; D013778:Tetrazolium Salts; D013844:Thiazoles
D000682:Amyloid; D016229:Amyloid beta-Peptides; D004396:Coloring Agents; D058228:Islet Amyloid Polypeptide; D010446:Peptide Fragments; D011328:Prions; D013778:Tetrazolium Salts; D013844:Thiazoles; C075222:amyloid beta-protein (1-42); C074636:amyloid beta-protein (25-35); C082632:prion protein (106-126); C022616:thiazolyl blue