Do aluminium and/or glutamate induce Alz-50 reactivity? A light microscopic immunohistochemical study.
Alzheimer's disease is thought to be characterized by conformational and phosphorylation changes in tau protein, leading to the formation of aggregations of paired helical filaments within neurons. Potential agents for inducing conformational changes in tau, namely aluminium and glutamate, were investigated in this study. Explant cultures of cortical neurons were established from embryonic day 17 rat fetuses. Cultures were exposed to aluminium, glutamate, aluminium/glutamate, aluminium/citric acid and citric acid (since aluminium is thought to enter cells via the transferrin receptor by complexing with citric acid) from 7-12 days in vitro. Control explants were exposed to basal medium only. On day 12, explants were paraffin-embedded. Four-six explants were serially sectioned per condition. For each explant, every 10th and adjacent 4 microm section were randomly selected and processed, with controls, for: (1) alcoholic morin histochemistry (to detect intracellular aluminium), (2) Perls' iron histochemistry (to control for the morin stain which also detects iron), (3) neurofilament immunohistochemistry (to estimate total neuronal number per explant) and (4) Alz-50 immunohistochemistry (to detect possible conformational changes in tau). The absolute number of stained/immunoreactive neurons was determined per explant. The percentage incidence was then determined per explant and averaged per condition. Explants in the aluminium conditions contained significant increases in the incidence of morin-positive aluminium containing neurons. There was also a significant increase in the incidence of Alz-50 positive neurons for the aluminium compared with control explants. These results suggest: (1) aluminium enters neurons and (2) aluminium alone induces possible conformational changes in tau as detected by the Alz-50 antibody, while aluminium combined with glutamate, or glutamate alone, do not.
Jones
K R
KR
Department of Anatomy and Structural Biology and the Neuroscience Research Centre, University of Otago Medical School, Dunedin, New Zealand.
Oorschot
D E
DE
D000535:Aluminum / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis / Q000378:metabolism*; D002452:Cell Count; D002540:Cerebral Cortex / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D019343:Citric Acid / Q000494:pharmacology; D046508:Culture Techniques; D004622:Embryo, Mammalian; D005419:Flavonoids / Q000032:analysis; D018698:Glutamic Acid / Q000494:pharmacology*; D007150:Immunohistochemistry; D009419:Nerve Tissue Proteins; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D011487:Protein Conformation; D051381:Rats; D017208:Rats, Wistar; D016875:tau Proteins / Q000032:analysis / Q000378:metabolism; D047309:Flavones
D000535:Aluminum; D000911:Antibodies, Monoclonal; D000941:Antigens; D019343:Citric Acid; D005419:Flavonoids; D018698:Glutamic Acid; D009419:Nerve Tissue Proteins; D016875:tau Proteins; C056167:Alzheimer's disease antigen; C008548:morin; D047309:Flavones