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The Journal of Neuroscience, August 1, 2002, 22(15):6394-6400

The Slow Axonal Transport of the Microtubule-Associated Protein Tau and the Transport Rates of Different Isoforms and Mutants in Cultured Neurons

Michelle A. Utton¹, James Connell¹, Ayodeji A. Asuni¹, Marjon van Slegtenhorst², Michael Hutton², Rohan de Silva³, Andrew J. Lees³, Chris C. J. Miller¹, and Brian H. Anderton¹

¹ Department of Neuroscience, Institute of Psychiatry, King's College London, London SE5 8AF, United Kingdom, ² Mayo Clinic Jacksonville, Jacksonville, Florida 32224, and ³ The Reta Lila Weston Institute of Neurological Studies, University College London, London W1T 4JF, United Kingdom

We demonstrate that the microtubule-associated protein tau, in the form of enhanced green fluorescent protein (EGFP) tau, is transported along axons of neurons in culture in the slow component of axonal transport with a speed comparable with that previously measured in vivo. It was demonstrated that the EGFP tag has no effect on transport characteristics, and the methodology enables slow transport rates of individual tau isoforms and tau mutants to be measured. We also expressed EGFP-tagged tau isoforms containing either three or four C-terminal repeats and zero or two N-terminal inserts in cultured neurons. No significant differences were found in the average rate of slow transport of the wild-type tau isoforms, suggesting that the exon 10 C-terminal repeat or the N-terminal inserts do not contain regions that play a significant regulatory role in axonal transport. Similarly, we found that missense mutations in tau have no noticeable effect on the rate of transport; hence their ability to cause neurodegeneration is by another mechanism other than that affecting the overall slow axonal transport of tau.

Key words: tau; tau isoforms; tau N-terminal inserts; tau C-terminal repeats; slow axonal transport; neuronal cultures; transfection; EGFP; FTDP-17

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Copyright © 2002 Society for Neuroscience  0270-6474/02/22156394-07$05.00/0

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