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Volume 16, Number 11,
Issue of June 1, 1996
pp. 3601-3619
Copyright ©1996 Society for Neuroscience
Tau Is Enriched on Dynamic Microtubules in the Distal Region of
Growing Axons
Received Feb. 8, 1996; revised March 5, 1996; accepted March 6, 1996.
Mark M. Black1,
Theresa Slaughter1,
Simon Moshiach1,
Maria Obrocka2, and
Itzhak Fischer2
1 Department of Anatomy and Cell Biology, Temple
University School of Medicine, Philadelphia, Pennsylvania 19140, and
2 Department of Neurobiology and Anatomy, Medical College
of Pennsylvania and Hahnemann University, Philadelphia, Pennsylvania
19129
It is widely held that tau determines the stability of microtubules
in growing axons, although direct evidence supporting this hypothesis
is lacking. Previous studies have shown that the microtubule polymer in
the distal axon and growth cone is the most dynamic of growing axons;
it turns over more rapidly and is more sensitive to microtubule
depolymerizing drugs than the polymer situated proximally. We reasoned
that if the stability of axonal microtubules is directly related to
their content of tau, then the polymer in the distal axon should have
less tau than the polymer in the proximal axon. We tested this
proposition by measuring the relative tau content of microtubules along
growing axons of cultured sympathetic neurons immunostained for tau and
tubulin. Our results show that the tau content of microtubules varies
along the axon, but in the opposite way predicted. Specifically, the
relative tau content of microtubules increases progressively along the
axon to reach a peak near the growth cone that is severalfold greater
than that observed proximally. Thus, tau is most enriched on the most
dynamic polymer of the axon. We also show that the gradient in tau
content of microtubules does not generate corresponding gradients in
the extent of tubulin assembly or in the sensitivity of axonal
microtubules to nocodazole. On the basis of these findings, we propose
that tau in growing axons has functions other than promoting
microtubule assembly and stability and that key sites for these
functions are the distal axon and growth cone.
Key words:
microtubule-associated proteins;
cytoskeleton;
axon
growth;
quantitative digital image analysis;
cultured sympathetic
neurons
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