Steroid hormones block amyloid fibril-induced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) formazan exocytosis: relationship to neurotoxicity.
Perhaps the most reproducible early event induced by the interaction of amyloid beta peptide (A beta) with the cell is the inhibition of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. We recently demonstrated that cytotoxic amyloid peptides such as A beta and human amylin inhibit cellular MTT reduction by dramatically enhancing MTT formazan exocytosis. We now show the following: (a) Insulin and glucagon, when converted to fibrils with beta-pleated sheet structure, induce MTT formazan exocytosis that is indistinguishable from that induced by A beta. NAC35, an amyloidogenic fragment of alpha-synuclein (or NACP), also induces MTT formazan exocytosis. (b) All protein fibrils with the beta-pleated sheet structure examined are toxic to rat hippocampal neurons. (c) Many sterol sex hormones (e.g., estradiol and progesterone) block amyloid fibril-enhanced MTT formazan exocytosis as well as MTT formazan exocytosis in control cells by acting at a common late step in the exocytic pathway. Steroids fail, however, to protect hippocampal neurons from acute amyloid fibril toxicity. These findings suggest that the ability to enhance MTT formazan exocytosis and to induce neurotoxicity are common biological activities of protein fibrils with beta-pleated sheet structure but that enhanced MTT formazan exocytosis is not sufficient for acute A beta neurotoxicity.
Liu
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Y
Cellular Neurobiology Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037-1099, USA.
Schubert
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D
D000682:Amyloid / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D005089:Exocytosis / Q000187:drug effects*; D005562:Formazans / Q000378:metabolism*; D005934:Glucagon / Q000494:pharmacology*; D012739:Gonadal Steroid Hormones / Q000494:pharmacology; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects; D007328:Insulin / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects; D009498:Neurotoxins / Q000494:pharmacology*; D010446:Peptide Fragments / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013261:Sterols / Q000494:pharmacology; D013552:Swine; D013778:Tetrazolium Salts / Q000378:metabolism*; D013997:Time Factors
D000682:Amyloid; D005562:Formazans; D012739:Gonadal Steroid Hormones; D007328:Insulin; D009498:Neurotoxins; D010446:Peptide Fragments; D013261:Sterols; D013778:Tetrazolium Salts; C079782:MTT formazan; D005934:Glucagon