Current Biology
Volume 4, Issue 12, December 1994, Pages 1077-1086
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Research Paper
Alzheimer's disease-like phosphorylation of the microtubule-associated protein tau by glycogen synthase kinase-3 in transfected mammalian cells

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Abstract

Background: Paired helical filaments (PHFs) are a characteristic pathological feature of Alzheimer's disease; their principal component is the microtubule-associated protein tau. The tau in PHFs (PHF-tau) is hyperphosphorylated, but the cellular mechanisms responsible for this hyperphosphorylation have yet to be elucidated. A number of kinases, including mitogen-activated protein (MAP) kinase, glycogen synthase kinase (GSK)-3α, GSK-3β and cyclin-dependent kinase-5, phosphorylate recombinant tau in vitro so that it resembles PHF-tau as judged by its reactivity with a panel of antibodies capable of discriminating between normal tau and PHF-tau, and by a reduced electrophoretic mobility that is characteristic of PHF-tau. To determine whether MAP kinase, GSK-3α and GSK-3β can also induce Alzheimer's disease-like phosphorylation of tau in mammalian cells, we studied the phosphorylation status of tau in primary neuronal cultures and transfected COS cells following changes in the activities of MAP kinase and GSK-3.

Results Activating MAP kinase in cultures of primary neurons or transfected COS cells expressing tau isoforms did not increase the level of phosphorylation for any PHF-tau epitope investigated. But elevating GSK-3 activity in the COS cells by co-transfection with GSK-3α or GSK-3β decreased the electrophoretic mobility of tau so that it resembled that of PHF-tau, and induced reactivity with eight PHF-tau-selective monoclonal antibodies.

Conclusion Our data indicate that GSK-3α and/or GSK-3β, but not MAP kinase, are good candidates for generating PHF-type phosphorylation of tau in Alzheimer's disease. The involvement of other kinases in the generation of PHFs cannot, however, be eliminated. Our results suggest that aberrant regulation of GSK-3 may be a pathogenic mechanism in Alzheimer's disease.

Section snippets

Background:

The mechanisms that result in neuronal loss in Alzheimer's disease are not understood. The discovery of several different mutations in the gene for the amyloid precursor protein in some pedigrees with familial Alzheimer's disease has given support to the ‘amyloid cascade’ hypothesis, in which the extracellular deposition of β-amyloid is an early pathogenic event (see [1], [2], [3] for reviews). A consensus has not yet been reached, however, on how deposition of β-amyloid in the brain results in

Discussion

In current hypotheses regarding the molecular pathology of Alzheimer's disease, aberrant amyloid precursor protein metabolism and the deposition of β-amyloid is placed as a primary pathogenic event (reviewed in [1], [2], [3]). It is becoming increasingly apparent, however, that cognitive deficits do not occur until dystrophic neurites containing PHFs and neurofibrillary tangles have developed [32], [33]. The transformation of tau to a hyperphosphorylated state as found in PHFs may thus

Primary neuronal cultures and transfection of COS-7 cells

Primary neuronal cultures derived from E17 (embryonic day 17) foetal rat brain cortices were prepared following standard protocols and plated at 3.2 times 105 cells cm−2 on dishes precoated with 10 mg ml−1 poly-L-lysine and 10 mg ml−1 laminin (Sigma) in glutamine-free culture medium comprising DMEM and Ham's F12 (4:1) supplemented with 100 mg ml−1 transferrin, 60 mM putrescine, 5 mg ml−1 insulin, 20 nM sodium selenite, 100 IU penicillin and 100 μg ml−1 streptomycin and 2% horse serum.

After 1

Acknowledgements

This research was funded by grants from the MRC, The Wellcome Trust and the Nuffield Foundation (to C.C.J.M.), a Wellcome Trust Programme grant (to B.H.A., C.C.J.M. and J-M.G), grants from the Commission of European Union and Glaxo Group research (to B.H.A) and a grant from The Wolfson Foundation (to C.C.J.M. and B.H.A). S.L. is supported by a Wellcome Trust Training Fellowship. We thank M. Weber, University of Virginia Health Sciences Center, Charlottesville, for kindly supplying a cDNA

Simon Lovestone, C. Hugh Reynolds, Donna Latimer, Daniel R. Davis, Brian H. Anderton, Diane Hanger, Sandrine Mulot and Christopher C.J. Miller (corresponding author), Department of Neuroscience, The Institute of Psychiatry, De Crespigny Park, Denmark Hill, London SE5 8AF, UK.

Jean-Marc Gallo, Department of Neurology, The Institute of Psychiatry, De Crespigny Park, Denmark Hill, London SE5 8AF, UK.

Betina Marquardt and Silvia Stabel, Max-Delbruck-Laboratorium, Carl-von-Linne-Weg 10, D-50829 Koln,

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  • Cited by (0)

    Simon Lovestone, C. Hugh Reynolds, Donna Latimer, Daniel R. Davis, Brian H. Anderton, Diane Hanger, Sandrine Mulot and Christopher C.J. Miller (corresponding author), Department of Neuroscience, The Institute of Psychiatry, De Crespigny Park, Denmark Hill, London SE5 8AF, UK.

    Jean-Marc Gallo, Department of Neurology, The Institute of Psychiatry, De Crespigny Park, Denmark Hill, London SE5 8AF, UK.

    Betina Marquardt and Silvia Stabel, Max-Delbruck-Laboratorium, Carl-von-Linne-Weg 10, D-50829 Koln, Germany.

    James R. Woodgett, Ontario Cancer Reseach Institute/Princess Margaret Hospital, 500 Sherbourne Street, Toronto, Ontario M4X 1K9, Canada.

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