Induction of Alzheimer-specific Tau epitope AT100 in apoptotic human fetal astrocytes.
In Alzheimer's and other neurodegenerative diseases, hyperphosphorylated tau accumulates in affected neuronal and glial cells in the form of paired helical filaments (PHFs). This tau binds antibody AT100, which recognizes the double phosphorylation site (Thr212/Ser214) that is not present in normal biopsy tau. In primary cultures, highly enriched (>98%) in astrocytes of human fetal brain, three polypeptides of 52, 64, and 70 kD showed immunoreactivity with tau antibodies against non-phosphorylated epitopes, accounting for 88, 12, and <1%, respectively, of the total reactivity. All three polypeptides were phosphorylated at the PHF-1 epitope but not at the epitopes Tau-1, 12E8, AT8, and AT100. Treatment of cultures with okadaic acid resulted in apoptosis characterized by the blebbing of the plasma membrane, condensation of nuclear chromatin, and fragmentation of the nucleus. This treatment also resulted in a 3- to 5-fold increase in the content of both tau protein and phosphorylation. The increases were observed in all phosphorylation sites examined, and included the AT100 site. The AT100 site has been proposed to be generated by protein kinase B/Akt and Cdc2. Since okadaic acid can induce an AD-like hyperphosphorylated state of normal tau in primary cultures of human brain cells, a simple cellular model is available permitting study of self-aggregation of tau and phosphorylation events characteristic of neurodegeneration.
Ksiezak-Reding
H
H
Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. reding@aecom.yu.edu
He
D
D
Gordon-Krajcer
W
W
Kress
Y
Y
Lee
S
S
Dickson
D W
DW
D017398:Alternative Splicing; D000544:Alzheimer Disease / Q000378:metabolism; D000906:Antibodies / Q000378:metabolism; D017209:Apoptosis; D001253:Astrocytes / Q000378:metabolism*; D001921:Brain / Q000196:embryology*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D053938:DNA Fragmentation; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D019319:Okadaic Acid / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D013997:Time Factors; D015854:Up-Regulation; D016875:tau Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D000906:Antibodies; D002843:Chromatin; D000939:Epitopes; D011518:Proto-Oncogene Proteins; D016875:tau Proteins; D019319:Okadaic Acid; C494918:AKT1 protein, human; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D017868:Cyclic AMP-Dependent Protein Kinases; D016203:CDC2 Protein Kinase