Journal of Alzheimer’s Disease 1 (1999) 329–351
IOS Press
Tau Protein in Normal and Alzheimer’s Disease
Brain: An Update*
Gail V.W. Johnson** and Judith A. Hartigan
Department of Psychiatry and Behavioral
Neurobiology, University of Alabama at
Birmingham, Birmingham, AL 35294-0017,
USA
ABSTRACT: Tau is a microtubule-associated protein
that, in a hyperphosphorylated form, comprises the
main component of the paired helical filaments and
neurofibrillary tangles found in Alzheimer’s Disease
(AD) brain. It is therefore important to understand the
normal functioning and processing of tau protein, and
the abnormal posttranslational processing of tau in AD
pathology. In 1996, Johnson and Jenkins reviewed the
literature on the biochemistry, function, and phos-
phorylation of tau in normal and AD brain. Since that
time, numerous publications have come out further
elucidating the properties of tau. The present review
updates the topics originally covered in the 1996
review, as well as presents a number of new topics.
For example, mutations in the tau gene have been
found in several non-AD, autosomal dominant
neurodegenerative disorders that exhibit extensive
neurofibrillary pathology. In addition, there is
increasing evidence that tau may be involved in signal
transduction, organelle transport, and cell growth,
independent of its microtubule-binding functions.
Taken together, the research reviewed here
demonstrates that tau is a very complex protein with
various functions that are intricately regulated. It is
clear that more research is required to completely
understand the functions and regulation of tau in
normal and AD brain.
*
This article is published with permission from the University
of Kentucky. It is also published in the online journal
Alzheimer’s Disease Review; www.coa.uky.edu/ADReview/
**
Corresponding author: Dr. Gail V.W. Johnson,
Department of Psychiatry, SC 1061, 1720 7th Avenue South,
University of Alabama at Birmingham, Birmingham, AL
35294-0017, USA, E-mail: gvwj@uab.edu
© University of Kentucky
329
KEYWORDS: Microtubule, phosphorylation, PHF,
kinase, phosphatase
INTRODUCTION
Tau is a microtubule-associated protein that is
selectively, but not exclusively (3,159),
expressed in the nervous system. In addition,
there is evidence of evolutionary conservation, as
tau-like proteins have been identified in
Caenorhabditis elegans (102). In Alzheimer’s
Disease (AD) brain tau is abnormally phos-
phorylated. This hyperphosphorylated form of
tau is the major component of the paired helical
filaments (PHFs) and neurofibrillary tangles
(NFTs) found in the AD brain. The mechanisms
underlying the abnormal phosphorylation and
accumulation of tau in AD are still under
investigation.
In 1996, Johnson and Jenkins (74) reviewed
the biochemistry, phosphorylation and function
of the tau protein in the normal and AD brain.
However, since that time, a significant number of
articles have been published further elucidating
the properties of tau. This review will present an
update on tau in normal and AD brain since the
time of the 1996 Johnson and Jenkins publication
(74). In addition to presenting new information
on the topics originally covered in the 1996
review, several new areas of research will be
discussed. For example, mutations in the tau
gene have been found in several families with
autosomal dominant forms of presenile non-AD
dementia. Also, there has been an explosion of
articles published on the role of glycogen
synthase kinase 3β (GSK-3β) in modulating tau
phosphorylation, and this work will be discussed.
The presence of tau in the cerebral spinal fluid
(CSF), which may be useful in the diagnosis of
AD, will also be reviewed. Finally, there is new
330 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
evidence to suggest some novel roles for tau,
such as being involved in signal transduction and
axonal outgrowth in a non-microtubule-
associated manner.
TAU AND PHF-tau:
STRUCTURE/FUNCTION
Historically, the C-terminal repeat regions of
the tau molecule were defined as microtubule
binding repeats (see Fig. 1). However, recent
studies have revealed that both the repeat regions
and the flanking domains are required for
efficient and productive tau-microtubule inter-
actions. In vitro, tau constructs containing only
the repeat regions of tau bound microtubules with
a relatively low affinity, however this region
efficiently promoted microtubule assembly and
stability. In contrast, the flanking domains of tau
bound microtubules with a relatively high
affinity, but did not effectively enhance micro-
tubule stability (49). These findings indicate that
the “targetting” flanking domains of tau are likely
responsible for positioning tau on the micro-
tubule surface, so that the “functional” repeat
domains can efficiently promote microtubule
stability and assembly. This model stresses the
importance of both the flanking and the repeat
domains for optimal tau-microtubule interactions.
In further support of this hypothesis, deletion of
the amino third of tau did not significantly alter
tau-microtubule interactions, however deletion of
∼ 30 amino acids within the proline-rich amino
terminal flanking domain drastically reduced the
affinity of the tau construct for microtubules
Fig. 1. Schematic Representation of the Tau Protein and the Location of PHF-Tau Phosphorylation Sites. Sites
phosphorylated in PHF-tau (107,176) are included on the top of the molecule, with Ser-Thr/Pro sites designated by
asterisks (*). Diagram is based on the 441 residue isoform (43). Flanking regions are based on (49).
(47). Using optical tweezers to bend individual
microtubules, Felgner (33) measured flexural
rigidity by their return movement to the resting
position in the presence of various tau constructs.
When full-length tau was bound, the flexural
rigidity of the microtubules was increased almost
3-fold over that observed with pure microtubules.
Tau constructs containing only the repeat regions
were not efficient at increasing the rigidity of the
microtubules, however addition of flanking
domains to the tau constructs greatly enhanced
their ability to decrease the flexibility of the
microtubules (33). In situ evidence of the role
these different domains of tau play in modulating
tau-microtubule interactions was obtained by
injecting different tau isoforms and constructs
into Chinese hamster ovary (CHO) cells, then
analyzing the association of tau with micro-
tubules (123). The results showed that the
presence of both the flanking regions and the
repeat domains was necessary for tau to
efficiently interact with microtubules in the cells,
further supporting the hypothesis that the
flanking and repeat domains play important
cooperative roles in modulating tau function
(123).
Taken together, these studies clearly indicate
that the regions of tau that have been referred to
as the “microtubule-binding domains” are the
important functional domains in terms of
promoting microtubule assembly and stability.
However, the flanking regions are essential for
targetting tau to the microtubules and contribute
to the function of tau as a microtubule stabilizing
protein (123). In addition, the flanking domains
of tau are hyperphosphorylated in AD brain, and
this likely contributes significantly to the
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
microtubule-binding incompetence of AD tau
(107).
A likely outcome of alterations in tau
phosphorylation is changes in the conformational
state of tau. Protein kinase A phosphorylation of
two serine residues in the flanking domains
causes a shift in electrophoretic mobility which is
indicative of a conformational change (16).
Further, mutation of these serine residues to
aspartate residues to mimic phosphorylation
results in conformational changes similar to those
caused by phosphorylation (89). In addition, it
has been demonstrated that the antibody Alz-50
recognizes tau in a specific conformation, and it
has been postulated that aberrant tau phos-
phorylation and/or polymerization into PHFs
induces the conformation that is recognized
selectively by Alz-50 (21). In vitro, PHF-tau
binds normally phosphorylated tau forming
filamentous aggregates, suggesting hyper-
phosphorylated tau may be able to facilitate PHF
formation by sequestering normal tau (2).
Although phosphorylation of specific sites
may facilitate the aggregation of tau into
filaments in AD brain, it is likely that other
factors contribute to this process. For example,
incubation of non-phosphorylated tau with
sulfated glycosaminoglycans, such as heparin or
heparan sulfate, resulted in the formation of PHF-
like structures (8,41,53,119). In addition, the
repeat region of tau was essential for the
induction of PHF formation (41). Heparin also
prevented tau from binding to microtubules and
promoted microtubule disassembly. Additionally,
there is some evidence to suggest that heparan
sulfate and tau may co-localize in NFT-
containing neurons in AD brain (41,78,120,
144,145). Considering this, Goedert et al. (41)
hypothesized that “an increase in sulphated
glycosaminoglycans within the cytoplasm of
nerve cells may trigger the hyperphosphorylation
of tau, destabilize microtubules and induce
assembly of PHFs”. However, further research is
required to elucidate the potential role of sulfated
glycosaminoglycans in AD pathology.
RNA also strongly enhances PHF assembly
and requires the formation of intermolecular
disulfide bridges involving the Cys322 residue
331
(numbering according to the longest human brain
tau isoform (43)) within the repeat domain of tau
(76). This cysteine was previously shown to play
a critically important role in the in vitro assembly
of different tau constructs into PHF-like
structures (134). Free fatty acids have been
shown to enhance filament formation of tau,
although the filaments were not PHF-like in
structure (170). Tissue transglutaminase (Tgase)
is a calcium-dependent cross-linking enzyme that
has been proposed to play a role in the function
of the insoluble NFTs in AD brain (31,138).
Tgase modifies human recombinant tau in vitro
resulting in the formation of insoluble
filamentous structures (4). Further, these fila-
mentous structures stained with an antibody to
Tgase just as strongly as PHFs isolated from AD
brain stained with the same antibody, suggesting
that cross-linking of tau by Tgase may be
involved in AD pathology (4). The self-
aggregation of tau into filaments can be inhibited
by the phenothiazine, methylene blue, and further
it reversed the proteolytic-resistance of PHFs by
interrupting the tau-tau interactions. This effect
of methylene blue was reportedly due to its
binding to the repeat region of tau thereby
inhibiting tau-tau interactions (171).
Recently it has been hypothesized that
differential expression of specific exons may
affect the ability of tau to form PHFs and NFTs.
Nelson et al. (113) cloned and sequenced tau-
encoding mRNA transcripts from the brains of
rhesus monkeys (which do not develop NFTs)
and of domesticated goat (which do develop
NFTs). The deduced amino acid sequence of
human tau has been previously reported (42,43).
The main difference between monkey, human
and goat tau mRNA transcripts was the presence
of exon 8 in monkeys. This exon was not found
in humans or goat. Exon 8 includes a highly
conserved Pro-Pro-Pro motif that would be
expected to introduce a sharp bend into the
protein structure. It was speculated that this bend
may sterically hinder phosphorylation and
polymerization, making animals that possess
exon 8 less susceptible to NFT formation (113).
In an earlier study using a polyclonal antibody to
exon 8, no exon 8 was detected in human tau
332 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
(22). However, there was also no exon 8 present
in rat or rabbit tau, which are species that do not
develop NFTs (22). Taken together, these data
indicate that the purported role of exon 8 in NFT
formation requires further study before any
conclusions can be made.
NOVEL FUNCTIONS
Tau is a microtubule-associated protein whose
historical role has been to promote microtubule
stability and assembly. However, recently a
number of papers have been published describing
functions of tau beyond its well-characterized
ability to modulate microtubule dynamics. Here
we will describe several of the novel functions
recently attributed to tau.
In 1996, Hwang et al. (63) demonstrated that
tau interacted with a key enzyme in the
phosphatidylinositol signalling pathway, phos-
pholipase C-γ (PLC-γ). The authors found that in
vitro, PLC-γ was maximally activated by a
combination of tau and arachidonic acid (63).
Further it was demonstrated that in human
neuroblastoma cells, tau selectively associates
with PLC-γ, and the PLC-γ that co-precipitates
with tau is enzymatically active (69). Ad-
ditionally, in vitro tau acts as a phosphoti-
dylinositol bisphosphate (PIP2) binding protein
like gelsolin, and it was suggested that the direct
binding of tau to PIP2 may make PIP2 a more
favorable substrate for PLC-γ (34). These data
strongly indicate that tau is likely to play a role in
signal transduction.
In addition to modulating the stability of
microtubules, tau is likely to regulate the motor-
protein mediated transport of organelles along
microtubules. Transfection of tau into COS cells,
which do not normally express tau, resulted in a
significant decrease in motor-based organelle
transport (129). In vitro, tau binds to one heavy
chain of the motor protein kinesin and the
presence of tau decreases the ability of micro-
tubules to stimulate the MgATPase activity of
purified kinesin (68). Additionally, activation of
protein kinase A in cultured sensory neurons
increased the transport of large organelles, an
event that coincided with increased tau
phosphorylation. Thus it was suggested that in
axons, regulation of tau phosphorylation may
modulate motor protein-dependent axonal
transport (129).
There is a growing body of evidence that tau
interacts either directly or indirectly with the
actin cytoskeleton and thereby plays a role in
regulating cell shape, motility and microtubule-
plasma membrane interactions. The M2 cell line,
derived from a malignant melanoma, lacks the
major actin-binding protein, ABP-280 (filamin),
and exhibits extensive membrane blebbing.
Microinjections of tau or MAP2 prevented bleb
formation and restored a normal phenotype (26).
Suppression of tau expression in cultured neurons
using antisense oligonucleotides results in a
significant reduction in growth cone area and
fillopodia number concomitant with considerable
changes in the phallodin staining of F-actin
(29). In addition, localization of tau to the distal
axon requires intact microfilaments (79). Al-
though these data indicate that tau may interact
with actin directly, it is more likely that tau is
interacting with actin through other proteins, as
in vitro MAP-2C efficiently promotes actin
gelatination, but tau does not (26). Additionally,
tau does not show a direct co-localization with
the microfilament network, indicating the exis-
tence of binding partners which could localize tau
to the actin cytoskeleton (79).
Neurite outgrowth is a specialized form of cell
motility, and it is clear that tau plays a role in this
process. Growing axons are very dynamic
structures and the stability of the microtubules
varies along the length of the axon. The distal
end of the axon is more sensitive to microtubule
depolymerizing drugs and the tubulin turnover is
more rapid than in the proximal end. Con-
sidering the role of tau as a microtubule
stabilizing protein, it was hypothesized that tau
would be more concentrated at the more stable,
proximal end and less concentrated at the more
dynamic, distal end. When this was tested in
cultured rat sympathetic neurons, Black et al.
(13) found the exact opposite. That is, tau was
most concentrated at the distal end of the axon, in
the growth cones (13). Similar results were
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
obtained using cultured rat hippocampal neurons
(79). Since in the distal regions of the growing
axon the microtubules are most dynamic, this
data strongly suggests that tau in the growing
axon has functions other than increasing
microtubule stability. One possibility is that the
tau in the developing neuron contains only three
repeat regions, since tau is developmentally
regulated, and this isoform has a reduced ability
to bind and stabilize microtubules.
In addition to concentration changes in tau
protein in the developing neuron, the phos-
phorylation state of tau was also found to differ
along the length of the growing axon (95). Using
fetal hippocampal cells, it was found that tau in
the growth cones was highly dephosphorylated at
the Tau-1 site as compared to the somatodendritic
compartment. A phosphorylation gradient was
evident, with a gradual change from phos-
phorylated to dephosphorylated tau from the
soma, through the axon, to the growth cone.
These studies again suggest that the function of
tau in the growing axon extends beyond its
classical microtubule stabilizing function because
in most cases dephosphorylation of tau increases
its affinity for microtubules.
IN VITRO PHOSPHORYLATION
STUDIES
In vitro, tau is phosphorylated by numerous
protein kinases (for earlier review see (75)), and
the list continues to grow (e.g., (117,172)). In
Table 1, sites on tau that have been reported to be
phosphorylated by specific protein kinases in
vitro are listed. It should be noted that although
many of the protein kinases and tau phosphory-
lation sites are indicated, this is not a
comprehensive list. Recently, members of the
stress-activated protein kinase (SAPK) family
have been shown to phosphorylate tau in vitro.
SAPK1γ (or Jun N-terminal kinase, JNK1),
SAPK2a (also known as p38, RK, CSBPs, Mpk2
or Mxi2), SAPK2b (or p38β), SAPK3 (also
known as ERK6 or p38γ) and SAPK4 all
phosphorylate tau at multiple Ser/Thr-Pro
epitopes, although SAPK3 and SAPK4 are
333
apparently the most efficient at phosphorylating
tau in vitro (39,125,126). Based on immunoblot
analysis using antibodies against specific
phospho-epitopes, the SAPKs have been shown
to phosphorylate several of the sites on tau that
are phosphorylated in AD brain (39,125,126).
Another proline-directed protein kinase that
phosphorylates tau and has received significant
attention lately is glycogen synthase kinase 3β
(GSK-3β, also called tau protein kinase I
(TPKI)). Earlier work demonstrated that tau is a
substrate of GSK-3β (51,93) and that prior
phosphorylation of tau by cdk5 (TPKII)
enhanced the rate and extent of subsequent
phosphorylation by GSK-3β (66,139). Speci-
fically, phosphorylation at Thr231 by GSK-3β was
enhanced 9-fold, which is a site that is hyper-
phosphorylated in PHF-tau. Additionally, phos-
phorylation of tau by GSK-3β has been shown to
reduce microtubule polymerization in vitro (161).
IN SITU PHOSPHORYLATION/DE-
PHOSPHORYLATION STUDIES
Recently numerous articles have been
published on the role of GSK-3β in tau phos-
phorylation in situ. Tau transiently expressed in
non-neuronal mammalian cell lines is not
phosphorylated significantly at sites found to be
phosphorylated in PHF-tau. However, when
GSK-3β is co-expressed with tau in these cells,
numerous Ser/Thr-Pro sites that are found in
PHF-tau become phosphorylated (65,92). Co-
expression of tau with GSK-3β also resulted in
less stable microtubules than when the cells were
transfected with only tau. Analogously, Wagner
et al. (167) transfected 3T3 and CHO cells with
tau and GSK-3β. Cells transfected with tau alone
displayed prominent bundles of microtubules.
However, co-transfection with tau and GSK-3β
led to increased tau phosphorylation and to a
decrease in microtubule bundling. The authors
also used a mutant form of tau in which five of
the sites known to be phosphorylated by GSK-3β
were converted to alanine to preclude
phosphorylation. Transfection of this mutant
form of tau alone resulted in microtubule
334 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain

Table 1
Reported In Vitro Phosphorylation Sites on Tau by Various Protein Kinases. Asterisks (*) represent sites on tau that
are phosphorylated in PHF-tau and by various protein kinases. The references cited in the table are as follows:
a, (107); b, (75); c, (85); d, (64); e, (117); f, (126); g, (125); h, (39)

bundling as did co-transfection of GSK-3β with
the mutant tau isoform (167). These results
suggest that the phosphorylation of tau by GSK-
3β reduces tau’s ability to organize microtubules.
Finally, Brownlees et al. (17) created transgenic
mice which expressed either wild-type GSK-3β
or a mutant form of GSK-3β that was more
active, and examined the phosphorylation state of
tau in the brains of the transgenic mice and non-
transgenic controls. The transgenics with the
highest level of GSK-3β activity showed elevated
tau phosphorylation using immunoblot analysis
with the AT8 antibody which recognizes tau
when Ser202 and Thr205 are phosphorylated (17).
Insulin and insulin growth factor-1 (IGF-1)
have been shown to modulate the activity of
GSK-3β (60). Prolonged activation of the insulin
or IGF-1 receptors often results in a down-
regulation of GSK-3β (60). In some systems this
has been shown to be due to activation of
phosphotidylinositol-3-kinase (PI-3-K). PI-3-K
activates Akt (also known as protein kinase B)
which phosphorylates GSK-3β on the Ser9
residue resulting in inactivation (25,60). Recent-
ly, insulin and IGF-1 were shown to decrease tau
phosphorylation in NT2N cells through a PI-3-
K — Akt — GSK-3β dependent pathway (60).
These data strongly indicate that tau is a
physiological substrate of GSK-3β.
In 1996, two reports clearly demonstrated that
in vitro lithium is an uncompetitive, selective
inhibitor of GSK-3β (81,151). In cultured NT2N
cells, lithium inhibited GSK-3β activity, and this
inhibition resulted in a decrease in tau
phosphorylation (59). Further, these investi-
gators found that lithium treatment reduced the
amount of tau that was associated with micro-
tubules. In SH-SY5Y cells, lithium treatment
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
reduced the phosphorylation of tau at some, but
not all, of the sites that were affected in NT2N
cells. However, in contrast to what was observed
in NT2N cells, lithium treatment did not alter the
amount of tau that was associated with
microtubules (174). Lithium also decreased tau
phosphorylation in cultured rat neurons (109).
Although GSK-3β is likely to be involved in
modulating tau phosphorylation, the data sugges-
ting a role for this kinase in the pathological
hyperphosphorylation of tau in AD brain are
equivocal. For example, immunoblot analyses of
supernatant and particulate fractions from AD
and control brains with antibodies to GSK-3β
revealed a 30% decrease in GSK-3β immuno-
reactivity in AD as compared to control brain.
Additionally there was a negative correlation
between GSK-3β levels and numbers of NFTs
(9). In contrast, Pei et al. (118) found a 50%
increase in GSK-3β levels in the post-
synaptosomal supernatant from AD brains as
compared to control brain using an indirect
ELISA. However, no increase in GSK-3β
enzyme activity in AD as compared to control
brains was observed (118). Considering these
disparate findings, it is clear that more research is
required to determine whether GSK-3β
contributes to tau hyperphosphorylation in AD.
A family of novel protein kinases named
MARK (MAP/microtubule affinity-regulating
kinase) have been implicated in phosphorylating
the critical Ser262 residue of tau (30). This
residue is located in the KXGS sequence of the
first repeat that is part of the microtubule-binding
domain of tau. Phosphorylation at this site is
elevated in AD (107) and Ser262 phosphorylation
significantly decreases tau-microtubule binding
(11,160). Transient expression of tau in CHO
cells that were stably transfected with the MARK
protein resulted in a disruption of the microtubule
array, suggesting that MARK phosphorylated tau
and decreased its ability to bind and stabilize
microtubules in situ (30). Interestingly, MARK
was also found to be inactivated by protein
phosphatase 2A (PP2A) (30), which is sig-
nificantly decreased in AD brain (45,46,147).
Therefore, the decrease in PP2A activity in AD
brain could contribute to the hyperphos-
335
phorylation of tau through two mechanisms.
Because tau is a substrate of PP2A (103,147),
decreased PP2A activity could directly result in
increased tau phosphorylation. In addition, a
decrease in PP2A activity could lead to increased
MARK activity, which may result in increased
Ser262 phosphorylation and lead to disruption of
microtubule assemblies.
There has also been a recent study that
indicates phosphorylation of Ser214 plays a
significant role in regulating the association of
tau with microtubules (64). Using 32P-labelled
cells and phosphopeptide mapping, it was
demonstrated that during mitosis there is a
significant increase in the phosphorylation of
Ser214, which is the site on tau which is pre-
ferentially phosphorylated by protein kinase A
(135). Phosphorylation of tau by protein kinase
A in vitro reduces its ability to nucleate
microtubules and promote microtubule assembly
(16). Because tau is an in situ substrate of
protein kinase A (35,91), it is likely that protein
kinase A is the protein kinase which phos-
phorylates tau on Ser214 in these mitotic LAN-5
and tau-transfected CHO cells (64), but further
validation is required.
It has also been suggested that increased
oxidative stress in AD brain may contribute to
the hyperphosphorylation of tau. However, the
majority of studies indicate that it is unlikely that
acute oxidative stress results in increased tau
phosphorylation. For example, both in rat brain
primary neuronal cultures (27) and in cultured
human neuroblastoma MSN cells (82), acute
oxidative stress resulted in tau dephos-
phorylation. However, enhanced tau phosphory-
lation and/or aggregation in response to low
level, chronic oxidative stress cannot be ruled
out.
Sadot et al. (128) provided evidence for a link
between the cholinergic signal transduction
system and the neuronal cytoskeleton. The
authors transfected rat PC12 cells with the m1
muscarinic acetylcholine receptor. They then sti-
mulated the cells with two acetylcholine agonists,
carbachol, and AF102B, which selectively
activates m1 receptors, and examined the
phosphorylation state of tau. Tau phos-
336 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
phorylation decreased at the Tau-1 and AT8 sites,
which include the Ser202 residue (128). This is
the first study to provide a link between the
cholinergic signal transduction pathway and tau
phosphorylation, and suggests that decreased
cholinergic activity could contribute to an
increase in tau phosphorylation.
Hall et al. (50) described a useful model in
which to study the cellular mechanisms under-
lying tau hyperphosphorylation. They injected
plasmids encoding human tau protein into
lamprey central neurons in vivo. In this model
the human tau was phosphorylated at the PHF-1
(Ser396/404) and Tau-1/AT8 (Ser199/202) sites.
Accumulations of tau were also observed and the
cells exhibited degenerative changes (50). This
model may be useful in studying tau
phosphorylation in situ and the relationship
between increased tau phosphorylation and
neurodegeneration.
In addition to protein kinases, protein
phosphatases play an important role in
modulating tau phosphorylation. Co-expression
of tau and SV40 small t antigen, a PP2A
inhibitor, in non-neuronal cells resulted in an
increase in tau phosphorylation at a number of
sites, including Ser199, Ser202, Thr205, Ser396 and
Ser404. This phosphorylation also resulted in a
loss of microtubule stability and the dissociation
of tau from the microtubules (147). In NT2N and
SH-SY5Y cells, depolymerization of micro-
tubules with nocadozole or stabilization of
microtubules with taxol results in the
dephosphorylation of tau at several Ser/Thr-Pro
epitopes by PP2A (103,174). Because a small
subpopulation of the total PP2A associates with
microtubules (146), it can be suggested that the
polymerization state of microtubules modulates
the phosphorylation state of tau indirectly
through its effects on PP2A. The calcium-
calmodulin dependent phosphatase calcineurin,
or PP2B, also regulates the phosphorylation state
of tau. Treatment of PC12 cells with maitotoxin
increases intracellular calcium levels and resulted
in tau dephosphorylation and calpain-mediated
tau degradation. Treatment of the cells with a
calcineurin inhibitor, FK502, prior to maitotoxin
resulted in a partial inhibition of tau dephos-
phorylation, but it completely inhibited calpain-
mediated tau proteolysis. These findings indicate
that dephosphorylation of tau by PP2B at specific
sites is necessary for calpain-mediated de-
gradation of tau (173).
In summary, there is a complex interplay
between the protein phosphatases and kinases in
the brain that modulate tau phosphorylation, and
it is likely that no one particular enzyme is
responsible for the hyperphosphorylated state of
tau in AD. The in situ studies cited here,
demonstrate the complexity of the systems in
living cells. Needless to say, there is much work
left to be done to understand the role of kinases
and phosphatases in modulating the phosphory-
lation state of tau in situ.
PHOSPHORYLATION OF PHF-tau
Recently, a number of new monoclonal tau
antibodies that recognize specific phospho-
epitopes or conformations have been described.
Antibodies directed against specific phospho-
epitopes in tau have proven to be excellent tools
in the elucidation of tau phosphorylation under
physiological and pathological conditions. A
summary of some of these numerous phospho-
specific tau antibodies is shown in Table 2. PHF-
9 specifically recognizes phosphorylated Ser404
(175), while AP422 recognizes phosphorylated
Ser422 (54). AD2 is another phosphorylation
dependent monoclonal antibody that recognizes
phosphorylated Ser396 and Ser404 (18). Hoffmann
et al. (58) describe two antibodies that are unique
in that they are dependent on the phosphorylation
of two residues. AT10 (also referred to as
AT100 in (176)) requires the simultaneous
phosphorylation of Thr212 and Ser214, while PHF-
27 requires the phosphorylation of the primary
site Thr231 and the subsite Ser235 (58). In
addition, both AT10 (AT100) and PHF-27 only
recognized PHF-tau and not fetal or biopsy
derived tau (58,176). These data indicate that
concurrent phosphorylation of tau on these sites
occurs in pathological conditions such as AD.
MC-1, a conformation dependent monoclonal
antibody, recognizes a tau conformation similar
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain 337

Table 2
Phosphoepitopes Recognized by Different Tau Antibodies. Indicates a phosphorylation pair, where
phosphorylation of both sites is required for optimal immunoreactivity. Indicates dephosphorylation. Stars
represent phosphorylation, with smaller stars indicating a subsequent minor phosphorylation site. The references
cited in the table are as follows: a, (154); b, (18); c, (54); d, (64); e, (176); f, (40); g, (55); h, (116); i, (100); j, (175);
k, (58); l, (143); m, (59); n, (141)

to that recognized by Alz-50 (71). As was
mentioned earlier, Alz-50 recognizes two
discontinuous segments of the tau molecule
(21). TG-3 is also a conformation dependent
monoclonal antibody that recognizes a tau
conformation found specifically in PHF-tau (72).
A subpopulation of vulnerable hippocampal
CA1 pyramidal neurons from AD brain stained
positively with PS 413, an antibody that
recognizes tau when Ser413 is phosphorylated
(143). Kimura et al. (80) investigated the
changes in tau phosphorylation as the NFTs
develop. Phosphorylation of sites Ser199, Ser202,
Ser409, and Ser422 occurs early in tangle develop-
ment and remain phosphorylated throughout the
NFT stages. In addition, Thr231 and Ser396 are
phosphorylated only in the more advanced stages
of NFTs (80). There is also evidence from AD
brain that casein kinase I selectively associates
with PHFs and may contribute to tau hyper-
phosphorylation (85).
One of the intriguing aspects of PHF-tau is
that it shares many of the same phosphorylated
sites as fetal tau. Fetal tau is also phosphorylated
to a significantly greater extent than control
human tau (77,169), which has led some to
hypothesize that a recapitulation of a fetal-like
state occurs in AD which is characterized by the
338 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
inappropriate expression of mitotic proteins. The
antibody MPM2 recognizes phosphoepitopes that
are found predominantly in mitotic cells (163).
Immunohistochemical analyses of AD hippo-
campus revealed that both MPM2 and AT8,
which recognizes tau when Ser202/Thr205 are
phosphorylated, stained NFTs (83,166) and co-
localized to pyramidal neurons (83). MPM2 also
reacts with PHF-tau, but not normal fetal or adult
rat tau (83). It has also been shown that the
presence of M-phase cdc2/cyclin B1 kinase
complex is elevated in AD brain compared to
controls, and the proteins were enriched in
neurons with NFTs and in neurons susceptible to
NFTs (165). Additionally, it has been established
that certain cell cycle proteins such as cyclin D,
cdk4, and proliferating cell nuclear antigen are
aberrantly expressed in AD brain (20). These
finding suggest that the inappropriate expression
of mitotic proteins may contribute to the
hyperphosphorylation of tau and the neuro-
degenerative process in AD.
It has been well-established that PHF-tau
exhibits significantly reduced electrophoretic
mobility and often presents as a triplet, with
relative molecular masses from 60 to 70 kDa
(88). Recently, the presence of an additional
∼ 74 kDa hyperphosphorylated tau form was
detected in the youngest and most severely
affected AD cases, and apparently corresponded
to the longest tau isoform found in brain.
Furthermore, based on isoelectric points, it was
determined that PHF-tau from older AD patients
was less phosphorylated than PHF-tau from
younger AD cases (140). These findings indicate
age of onset and duration of the disease are
important factors to consider when evaluating the
characteristics of PHF-tau.
OTHER TAU MODIFICATIONS
Lipid peroxidation, and the formation of its
aldehydic by-product 4-hydroxynonenal (HNE),
occur in response to oxidative stress and have
been implicated in the modifications of tau in AD
brain. In a preliminary study, treatment of
cultured rat hippocampal cells with HNE resulted
in tau being modified in such a way that it could
not be efficiently dephosphorylated (98). In P19
neuroglial cultures, HNE was found to stimulate
the formation of ubiquitinated cross-linked
proteins, including tau, but was also a potent
cytotoxin (105). Although these studies are
intriguing, future work is necessary to determine
if lipid peroxidation or HNE-linked modifications
contribute to the tau pathology in AD brain.
Glycation of tau is another potential
modification that may be involved in the
formation of PHFs. As reported previously, tau
is a good candidate for glycation, and PHF-tau
was found to be glycated, while normal tau was
not (74). Glycation is the non-enzymatic ad-
dition of a reducing sugar to a protein, often on a
lysine residue, and is a modification that often
occurs in long-lived proteins during the aging
process (10). Nacharaju et al. (112) characterized
the sites on tau that are glycated in vitro to
determine if the repeat regions are involved.
Using the longest isoform of tau, it was
demonstrated that six sites within the repeat
regions (Lys259, 280, 281, 347, 353, and 369) were gly-
cated (112). Since this area of the tau molecule is
important for binding and stabilizing micro-
tubules, glycation here could disrupt normal tau
function and contribute to the formation of PHFs
and NFTs. However, because it is long-lived,
lysine-rich proteins that are modified, it is
conceivable that glycation of PHF-tau occurs
subsequent to PHF and NFT formation. There is
also preliminary data suggesting that the levels of
oxidized products from non-enzymatic glycation
are not elevated in AD brain compared to
controls (136). Therefore, the modification of
proteins by glycation in AD brain may be a
selective process and not the result of global
changes.
Tau is apparently modified by glycosylation.
Ser(Thr)-O-linked N-acetylglycosamination is a
dynamic and abundant posttranslational modifi-
cation that can be reciprocal with Ser/Thr phos-
phorylation. In an initial study, bovine tau was
found to be modified by O-linked glycosylation
at multiple sites, and it is likely that this
modification may play a role in modulating tau
function (7). The modification of PHF-tau, but
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
not normal tau, by N-linked glycosylation has
also been reported (168). In contrast to O-linked
glycosylation, N-linked glycosylation usually
occurs co-translationally and is a considerably
less dynamic modification. Interestingly, degly-
cosylation of PHFs converted them into straight
filaments and therefore it was suggested that N-
linked glycosylation may contribute to the
maintenance of PHF-structure (168).
Aβ, CALCIUM HOMEOSTASIS AND
TAU HYPERPHOSPHORYLATION
Aβ constitutes the core of the extracellular
neuritic plaques, another neuropathic hallmark of
AD in addition to the NFTs. Aβ is a 39-43 amino
acid peptide that results from the cleavage of the
695-770 amino acid amyloid precursor protein
(APP) (for a review see (52,96,137). Because
neuritic plaques and NFTs occur in AD brain, the
interrelationship between the two major
components of these structures, tau and Aβ, is
being examined. Diffuse and dense neuritic
plaques in AD brain immunohistochemically
label with antibodies against PHF-tau (131) and
PHF-tau positive neurites are present in and
around all subtypes of plaques (153). Converse-
ly, antibodies to an epitope between residues 713
and 723 of APP770 consistently labeled PHFs in
the AD brain (38). In vitro, a peptide segment
corresponding to residues 713 to 730 of APP770
(which is in the C-terminus region) bound to tau
and formed dense fibrillary assemblies containing
both tau and APP (38). Domains of APP that are
C-terminal of the Aβ-containing segment bind to
tau and tubulin in vitro (67), and the non-
amyloidogenic Aβ1–28 also bound directly to
tubulin (67). These data indicate that direct
interactions between tau and specific segments of
APP, including the Aβ domain, may occur both
normally and/or pathologically in AD brain.
There have also been numerous studies
examining the possible effects of Aβ on tau
phosphorylation. Treatment of fetal rat hippo-
campal and human cortical neurons with fibrillar
Aβ resulted in an increase of tau phosphorylation
at Ser202/Thr205 and Ser396/Ser404 (19). The phos-
339
phorylation of tau in response to Aβ treatment
resulted in a loss of microtubule binding capacity
and accumulation of tau in the somatodendritic
compartment (19). Similarly, treatment of the
hybrid septal cell line, SN56, with Aβ1–40 resulted
in tau phosphorylation at Ser199/202 and Ser396, and
cell toxicity. However, treatment with Aβ1–28,
which is non-amyloidogenic, had no significant
effects on tau phosphorylation (87). Interesting-
ly, it was recently reported that neuronal cell
cultures were protected from Aβ toxicity by
taxol, a microtubule-stabilizing drug (104).
There is also some data to indicate that increased
Aβ deposition in animal models also results in
increased tau phosphorylation. For example, a
recent study in transgenic mice overexpressing
human APP containing either the Swedish double
mutation at positions 670/671 alone or in
combination with the London V717I mutation,
demonstrated that the dystrophic neurites that
developed around the plaques stained positive for
specific phosphorylated forms of tau. These
immunohistochemical findings were corraborated
by immunoblot analysis which also revealed an
increase in tau phosphorylation (152).
One possible mechanism by which Aβ may
induce increases in tau phosphorylation is by
perturbing calcium homeostasis which would
result in inappropriate increases in the activities
of certain protein kinases contributing to tau
hyperphosphorylation. Further, it has been
hypothesized that Aβ-induced perturbation of
calcium homeostasis could make the cells more
vulnerable to other potentially “toxic” stimuli,
including glutamate. Treatment of human cere-
bral cortical or rat hippocampal cell cultures with
Aβ resulted in elevated intracellular calcium
levels over those of resting cells. Addition of Aβ
also made the cells more vulnerable to
neurotoxicity induced by glutamate, kainate or N-
methyl-D-aspartate (NMDA) (97,99). Interes-
tingly, rat hippocampal cells were protected
against Aβ induced calcium perturbation by basic
fibroblast growth factor (bFGF), and therefore it
was suggested that bFGF may be involved in
stabilizing intracellular calcium levels (99).
Although treatment with Aβ may result in
increases in intracellular calcium, this may not be
340 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
the mechanism by which it induces the putative
increases in tau phosphorylation. For example,
treatment of human neuroblastoma cells with
either a calcium ionophore or Aβ25–35 resulted in
an increase in intracellular calcium levels and
increased tau phosphorylation at specific epitopes
(142). The increase in tau phosphorylation elicited
by the calcium ionophore was blocked by the
calcium chelator EGTA, however the Aβ25–35
induced increase in tau phosphorylation was not,
suggesting that it was not mediated by calcium
(142). Further, there is some evidence to suggest
that Aβ does not increase intracellular calcium
concentrations. For example, Aβ25–35 treatment of
primary rat cortical neurons (27) and Aβ1–42
treatments of human neuron-like NT2N cells (37)
did not result in an increase in intracellular
calcium concentrations. The conflicting nature of
these findings clearly indicates the need for
additional studies to clarify the involvement of
calcium-mediated events in Aβ-induced increases
in tau phosphorylation.
Another possible mechanism by which Aβ may
induce increases in tau phosphorylation is by
activation of GSK-3β. GSK-3β is one of the few
protein kinases shown to phosphorylate tau in situ
(17,51,59,60,161,167). Exposure of rat hippo-
campal cell culture to Aβ resulted in GSK-3β
activation and increased tau phosphorylation
(61). In addition, activation of GSK-3β by Aβ
may contribute to neuronal death. Treatment of
rat hippocampal cells with GSK-3β antisense
oligonucleotides prevented Aβ-induced neuronal
death (156). Furthermore, it was demonstrated
that exposure of rat hippocampal cells to Aβ25–35
results in the inactivation of phosphatidylinositol-3
(PI-3) kinase (155). PI-3 kinase activates Akt
which phosphorylates GSK-3β on Ser9 leading to
inactivation and decreased tau phosphorylation
(60). Therefore, inactivation of PI-3 kinase by Aβ
would result in an increase of GSK-3β activity and
subsequent increase in tau phos-phorylation.
These findings have lead to the hypothesis that Aβ
decreases PI-3 kinase activity thus disinhibiting
GSK-3β which in turn results in increased tau
phosphorylation, impaired axonal transport and
ultimately somatodendritic tau accumulation and
neuronal death (61, 155,156).
TAU IN OTHER
NEURODEGENERATIVE DISEASES
Hyperphosphorylated forms of tau occur in
many neurodegenerative diseases besides AD,
and in almost all cases accumulates as
filamentous lesions. The presentation of the
hyperphosphorylated tau shows some variability
between the diseases, both in terms of
electrophoretic mobility (i.e., migrating as two,
three, or four bands) and in the ultrastructural
characteristics of the filaments (e.g., straight
versus paired helical, filament dimensions, etc.)
(32,84,101,132). However, it has been hypo-
thesized that because of the clinical and
pathological overlap among neurodegenerative
disorders with extensive tau pathology, such as
AD, corticobasal degeneration (CBD), progres-
sive supernuclear palsy (PSP), and Guam amyo-
trophic lateral sclerosis/Parkinsonism-dementia
complex (ALS/PDC), that it is likely that they
share some etiological correlates (32). Interes-
tingly, there is growing evidence that mutations
in the tau gene may be a contributing factor in
several neurodegenerative diseases with ex-
tensive tau pathology. For example, intronic
polymorphisms in the tau gene may be involved
in PSP (23,57).
Although tau mutations have been implicated
in certain familiar forms of PSP (23,57), strong
evidence for a role of tau mutations in certain
families with autosomal dominant forms of
presenile frontotemporal dementia (FTD) has
recently been described. Several pedigrees of
FTD have been linked to chromosome 17q21
(12,56,111,149) which is also the location of the
tau gene (1). This subset of frontotemporal
dementias is collectively referred to as “fronto-
temporal dementia and Parkinsonism linked to
chromosome 17” (FTDP-17). Several of these
families have abundant widespread tau-positive
fibrillary deposits in both neurons and glia
(12,111,124,149). Although abundant neuro-
fibrillary inclusions are present in these FTDs,
amyloid plaques were virtually absent. In recent
reports, specific mutations in the tau gene have
been shown to occur in a number of these
families with FTDP-17 (122,150). Mutations
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
have been found in the tau introns (62,150),
including the 5' splice site of exon 10 (62), as
well as in the coding region of the tau gene (122).
It was hypothesized that mutations of the tau
introns may be responsible for the increase in
synthesis of the larger isoforms of tau and result
in abnormal tau aggregation (62,150). Several
mutations occurred within the microtubule-
binding domains of tau, and therefore could
disrupt tau-microtubules interactions and thus
contribute to tau aggregation and neuro-
degeneration (62,150).
It is intriguing that in these FTDP-17 diseases,
as well as a number of other neurodegenerative
diseases including CBD, PSP and ALS/PDC,
there is extensively neurofibrillary pathology
without abundant amyloid plaques (148). These
data suggest that in AD the amyloid plaques and
NFTs are independent lesions and one is not the
sole “cause” of the other. This is not to rule out
the possibility that the consequences of the
presence of one lesion may potentiate the
development of the other. For example, Aβ
could, through some mechanism, contribute to
the increase in tau phosphorylation (e.g. see
preceding section). Indeed, in one of the amyloid
precursor protein transgenic mice models,
increased tau phosphorylation was observed,
especially around the amyloid plaques (152),
suggesting that Aβ deposition may facilitate the
hyperphosphorylation of tau. Therefore, further
study of the interrelationship between Aβ and tau
pathologies is essential.
TAU IN THE CEREBROSPINAL FLUID
The primary mechanism by which an
antemortem diagnosis of AD is made is through
the use of neuropsychological analysis. Although
neuropsychological testing is extremely useful,
definitive diagnosis of AD based on objective
biological markers would greatly facilitate the
early identification of AD patients. The ability to
accurately identify AD patients early in the
course of the disease will become even more
imperative as effective treatments of the disease
become available. Presently, it is not until
341
autopsy that AD can be diagnosed conclusively.
Therefore, the presence of biomarkers specific
for AD have been sought. In 1993, Vander-
meeren et al. (162) developed a sensitive sand-
wich enzyme-linked immunosorbent assay to
detect the low levels of tau found in the CSF, and
with this assay found an increase in CSF tau
levels in AD patients (162). Shortly thereafter,
Mandelkow and Mandelkow (94) suggested that
tau levels in the CSF may be used as a biomarker
for AD, when they also found an increase of tau
in the CSF of AD patients (94). Further, the tau
found in the CSF of both controls and AD
patients is not full length tau, but a proteolytic
product containing the N-terminal end of the
molecule (73). There are now numerous articles
on tau in the CSF, and the overwhelming
consensus is that tau levels in the CSF of AD
patients are significantly increased over the CSF
tau levels of normal controls (e.g. 36,70,106,108,
110,127,158,164). CSF tau levels from AD
patients were also elevated over those with multi-
infarct dementia (158), cerebrovascular disease
dementia (106), non-AD dementia (164), and
mild cognitive impairment and other dementing
diseases (15). Galasko et al. (36) found an
increase in CSF tau levels from mildly impaired
patients with AD, suggesting that elevated levels
of CSF tau occur very early in the disease.
However, one study reported that CSF tau levels
from AD patients were as high as those from
patients with dementia with Lewy bodies
(5/6). Elevated levels of tau in CSF were also
found in patients suffering from Creutzfeldt-
Jakob Disease (CJD) (115). Blennow et al. (14)
found considerable overlap in CSF tau levels
with AD patients and those with other forms of
dementia including vascular dementia and frontal
lobe dementia (14). These studies indicate that
elevated CSF tau levels alone cannot be used to
conclusively diagnose AD.
In addition to increases in CSF tau, decreases
in Aβ1–42 peptide in the CSF may also provide
support for a diagnosis of AD. CSF levels of
soluble APP have been found to be decreased in
AD patients as compared to controls (86,121).
However, a more consistent and reliable
difference has been found in CSF Aβ1–42
342 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
levels. The CSF levels of Aβ1–42 have been
found to be significantly decreased in AD
patients compared to depressed control subjects
(133) and normal controls (108,157). The
results on the Aβ1–40 segment are mixed, with
some showing no change in AD as compared to
controls (157) and others showing decreased
CSF Aβ1–40 in AD over depressed controls
(133). These data indicate that increased CSF
tau levels in combination with decreased CSF
Aβ1–42 levels may be useful indicators in the
diagnosis of AD.
Apolipoprotein E alleles have also been
detected in the CSF. A full discussion of the
role of apolipoprotein E is beyond the scope of
this paper. Suffice it to say that the presence of
the apolipoprotein E-4 allele (apoE4) is a
genetic risk factor for sporadic and familiar late-
onset AD (24,130). Numerous researchers have
measured the levels of both tau and apoE4 in the
CSF to determine whether there is a connection
between the two. To date, the results of these
experiments are mixed, with some authors
finding that increased CSF tau levels are
correlated with increased CSF apoE4 levels
(15,44,90) and others finding no relationship
between CSF tau levels and CSF apoE4 levels
(6,108,114).
Another area where the results are mixed is in
whether the CSF tau levels correlate with the
severity of cognitive impairment or dementia.
Tato et al. (158) found that CSF tau levels
correlated positively with the evolution of the
disease and negatively with a mental state
examination. However, others have found no
correlation between CSF tau levels and degree
of cognitive impairment or severity of dementia
(110,114,127).
At this point, it is clear that elevated CSF tau
levels are insufficient to make a definitive
diagnosis of AD. The combination of elevated
CSF tau levels and decreased CSF Aβ1–42 levels
increases the reliability of an AD diagnosis but
is still not sufficiently definitive to make a
conclusive diagnosis. The ability to use CSF tau
levels in conjunction with other markers may
increase the accuracy of a diagnosis of AD,
however more studies are clearly necessary.
CONCLUSIONS AND SPECULATIONS
It is becoming increasingly evident that
classification of tau simply as a microtubule
binding and stabilizing protein is insufficient.
Recent work strongly indicates that tau is
multifunctional and likely to play a significant
role in a variety of neuronal functions. In order
to identify the mechanisms responsible for the
abnormal accumulation of tau in AD and other
neurodegenerative diseases, a clearer under-
standing of tau’s functions, and the modifications
which regulate these functions, is required.
Tau is a highly phosphorylated protein, and it
is readily evident that site-specific phos-
phorylation of the molecule is a very dynamic
process. Recent studies have begun to provide
significant insight into the protein kinases and
phosphatases that modulate the phosphorylation
state of tau in situ. However, much remains to be
learned about the dynamic interplay between
specific protein kinases and phosphatases in
normal and pathological conditions, as well as
the identity of these enzymes which modulate the
in vivo phosphorylation state of tau. It is also
evident that phosphorylation is not the only
regulatory posttranslational modification of tau.
Glycosylation, as well as other modifications, are
likely to regulate tau function and localization.
Elucidating the posttranslational processing of
tau is critically important to our understanding of
the involvement of tau in the neuropathology of
AD.
The recent findings that mutations in the tau
gene have been linked to several autosomal
dominant FTDP-17’s is particularly exciting. In
at least two of the pedigrees, fibrillary deposits
composed of hyperphosphorylated tau are
prevalent but there is a marked absence of Aβ
deposits. These findings indicate that abnor-
malities in the processing and function of tau are
likely to contribute to the neurodegenerative
process. The studies strongly suggest that in AD
and these other neurodegenerative diseases, the
abnormal modification of tau and eventual
aggregation into neurofibrillary lesions is not just
an “outcome” of the neuronal death process, but
rather plays a critical important role in the
 G.V.W. Johnson and J.A. Hartigan / Tau Protein in Normal and Alzheimer’s Disease Brain
neurodegenerative process. However, before any
definitive conclusions can be reached, it is clear
that further studies are required.
ACKNOWLEDGMENTS
Our research was supported by grants from the
NIH (AG12396, AG06569, NS27538 and
NS35060).
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