Selective Phosphorylation of Adult Tau Isoforms in Mature Hippocampal Neurons Exposed to Fibrillar Aβ

doi:10.1006/mcne.1997.0615

Molecular and Cellular Neuroscience

Volume 9, Issue 3, 1997, Pages 220-234

https://doi.org/10.1006/mcne.1997.0615 Get rights and content

Abstract

How senile plaques and neurofibrillary tangles are linked represents a major gap in our understanding of the pathophysiology of Alzheimer's disease. We characterized a hippocampal neuronal culture system in which tau undergoes maturationin vivo;rat neurons maintained in culture for more than 3 weeks replicated the splicing and phosphorylation changes that tau undergoes upon maturationin situ.Using this model system, we induced an Alzheimer-like neuritic dystrophy following the application of fibrillar β-amyloid. The dystrophy consisted of focal distortions and swellings within the neurites and an altered phosphorylation of the adult tau isoforms. Fibrillar β-amyloid induced the concomitant activation of MAP kinase and GSK3 β. The aberrant activation of several signaling pathways may lead to the abnormal phosphorylation of tau and neuritic degeneration.

References (55)

G.T. Bramblett et al.
Abnormal tau phosphorylation at Ser 396 in Alzheimer's disease recapitulates development and contributes to reduce microtubule binding
Neuron
(1993)
J. Busciglio et al.
β-Amyloid fibrils induce tau phosphorylation and loss of microtubule binding
Neuron
(1995)
H. Eldar-Finkelman et al.
Inactivation of glycogen synthase kinase-3 by epidermal growth factor is mediated by mitogen-activated protein kinase/p90 ribosomal protein S6 kinase signaling pathway in NIH/3T3 cells
J. Biol. Chem.
(1995)
A. Ferreira et al.
Microtubule formation and neurite growth in cerebellar macroneurons which develop in vitro: evidence for the involvement of the microtubule-associated proteins, MAP-1a, HMW-MAP-2 and tau
Dev. Brain Res.
(1989)
M. Goedert et al.
Multiple isoforms of human microtubule-associated protein tau: Sequences and localization in neurofibrillary tangles of Alzheimer's disease
Neuron
(1989)
M. Goedert et al.
p42 MAP kinase phosphorylation sites in microtubule-associated protein tau are dephosphorylated by protein phosphatase 2A
FEBS Lett.
(1992)
D.P. Hanger et al.
Glycogen synthase kinase 3 induces Alzheimer's disease-like phosphorylation of tau: Generation of paired helical filament epitopes and neuronal colocalization of the kinase
Neurosci. Lett.
(1992)
S. Kobayashi et al.
A cdc-related kinase PSSALRE/cdk5 is homologous with the 30 kDa subunit of tau protein kinase II, a proline-directed protein kinase associated with microtubules
FEBS Lett.
(1993)
K.S. Kosik et al.
Epitopes that span the tau molecule are shared with paired helical filaments
Neuron
(1988)
K.S. Kosik et al.
Developmentally regulated expression of specific tau sequences
Neuron
(1989)

M.D. Ledesma et al.

Implication of brain cdc2 and MAP kinases in the phosphorylation of tau proteins in Alzheimer's disease

FEBS Lett.

(1992)

O.H. Lowry et al.

Protein measurements with the folin phenol reagent

J. Biol. Chem.

(1951)

E.-M. Mandelkow et al.

Glycogen synthase kinase 3 and the Alzheimer's disease-like state of microtubule-associated protein tau

FEBS Lett.

(1992)

E.S. Matsuo et al.

Biopsy-derived adult human brain tau is phosphorylated at many of the same sites as Alzheimer's disease paired helical filament tau

Neuron

(1994)

R.L. Neve et al.

Identification of cDNA clones for the human microtubule-associated protein tau, and chromosomal localization of the genes for tau and microtubule associated protein 2

Mol. Brain Res.

(1986)

H.K. Paudel et al.

Brain proline-directed protein kinase phosphorylates tau on sites that are abnormally phosphorylated in tau associated with Alzheimer's paired helical filaments

J. Biol. Chem.

(1993)

C.J. Smith et al.

Tau isoform expression and phosphorylation state during differentiation of cultured neuronal cells

FEBS Lett.

(1995)

C. Sutherland et al.

The α-isoform of glycogen synthase kinase-3 from rabbit skeletal muscle is inactivated by p70 S6 kinase or MAP kinase-activated protein kinase-1 in vitro

FEBS Lett.

(1994)

S. Temple et al.

Clonal analysis of oligodendrocyte development in culture: Evidence for a developmental clock that counts cell divisions

Cell

(1986)

R. Vulliet et al.

Proline-directed phosphorylation of human tau protein

J. Biol. Chem.

(1992)

Q.M. Wang et al.

Use of a synthetic peptide as a selective substrate for glycogen synthase kinase 3

Anal. Biochem.

(1994)

M. Arioki et al.

Tau protein kinase II is involved in the regulation of the normal phosphorylation state of tau protein

J. Neurochem.

(1993)

K. Bauman et al.

Abnormal Alzheimer's like phosphorylation of tau protein by cyclin-dependent cdk2 and cdk5

FEBS Lett.

(1993)

A. Bensadaun et al.

Assay of protein in the presence of interfering material

Anal. Biochem.

(1976)

J.E. Bottenstein et al.

Growth of a rat neuroblastoma cell line in serum-free supplemented media

Proc. Natl. Acad. Sci. USA

(1979)

J.P. Brion et al.

Developmental changes in τ phosphorylation: Foetal τ is transiently phosphorylated in a manner similar to paired helical filament τ characteristic of Alzheimer's disease

J. Neurochem.

(1993)

A. Caceres et al.

Inhibition of neuronal polarity by tau antisense oligonucleotides in primary cerebellar neurones

Nature

(1990)

Cited by (174)

The hidden players: Shedding light on the significance of post-translational modifications and miRNAs in Alzheimer's disease development
2023, Ageing Research Reviews
Alzheimer’s disease (AD) is the most prevalent, expensive, lethal, and burdening neurodegenerative disease of this century. The initial stages of this disease are characterized by a reduced ability to encode and store new memories. Subsequent cognitive and behavioral deterioration occurs during the later stages. Abnormal cleavage of amyloid precursor protein (APP) resulting in amyloid-beta (Aβ) accumulation along with hyperphosphorylation of tau protein are the two characteristic hallmarks of AD. Recently, several post-translational modifications (PTMs) have been identified on both Aβ as well as tau proteins. However, a complete understanding of how different PTMs influence the structure and function of proteins in both healthy and diseased conditions is still lacking. It has been speculated that these PTMs might play vital roles in the progression of AD. In addition, several short non-coding microRNA (miRNA) sequences have been found to be deregulated in the peripheral blood of Alzheimer patients. The miRNAs are single-stranded RNAs that control gene expression by causing mRNA degradation, deadenylation, or translational repression and have been implicated in the regulation of several neuronal and glial activities. The lack of comprehensive understanding regarding disease mechanisms, biomarkers, and therapeutic targets greatly hampers the development of effective strategies for early diagnosis and the identification of viable therapeutic targets. Moreover, existing treatment options for managing the disease have proven to be ineffective and provide only temporary relief. Therefore, understanding the role of miRNAs and PTMs in AD can provide valuable insights into disease mechanisms, aid in the identification of biomarkers, facilitate the discovery of novel therapeutic targets, and inspire innovative treatments for this challenging condition.
The formation of small aggregates contributes to the neurotoxic effects of tau<inf>45-230</inf>
2022, Neurochemistry International
Citation Excerpt :
The microtubule-associated protein (MAP) tau plays an important role in several neurodegenerative diseases collectively known as tauopathies (reviewed by Iqbal et al., 2005; Ding and Johnson 2008; Hernandez and Avila, 2008; Mandelkow and Mandelkow, 2012; Iqbal et al., 2016; Guo et al., 2017). In these diseases, hyperphosphorylated tau is abundant in affected brain areas either in monomeric or aggregated forms (Grudke-Iqbal et al., 1986; Kosik et al., 1986; Wood et al., 1986; Takashima et al., 1993; Ferreira et al., 1997; Gamblin et al., 2003; Rapoport et al., 2002; Huang et al., 2016; Quinn et al., 2018). The tau45-230 fragment is also easily detectable in tauopathies (Ferreira and Bigio, 2011).
Intracellular deposits of hyperphosphorylated tau are commonly detected in tauopathies. Furthermore, these aggregates seem to play an important role in the pathobiology of these diseases. In the present study, we determined whether the recently identified neurotoxic tau_45-230 fragment also formed aggregates in neurodegenerative disorders. The presence of such aggregates was examined in brain samples obtained from Alzheimer's disease (AD) subjects by means of Western blot analysis performed under non-denaturing conditions. Our results showed that a mixture of tau_45-230 oligomers of different sizes was easily detectable in brain samples obtained from AD subjects. Our data also suggested that tau_45-230 oligomers could be internalized by cultured hippocampal neurons, mainly through a clathrin-mediated mechanism, triggering their degeneration. In addition, in vitro aggregation studies showed that tau_45-230 modulated full-length tau aggregation thereby inducing the formation of smaller, and potentially more toxic, aggregates of this microtubule-associated protein. Together, these data identified alternative mechanisms underlying the toxic effects of tau_45-230.
Altered Cytoskeletal Composition and Delayed Neurite Elongation in tau<inf>45–230</inf>-Expressing Hippocampal Neurons
2019, Neuroscience
Calpain-mediated tau cleavage into the neurotoxic tau_45–230 fragment plays an important role in Alzheimer's disease (AD). This tau fragment accumulates mainly in the cytoplasm of degenerating neurons. However, subcellular localization studies indicated that a pool of tau_45–230 associates with the cytoskeleton in hippocampal neurons. In the present study, we assessed whether such localization could underlie tau_45–230 neurotoxic effects. Quantitative Western blot analysis showed decreased levels of full-length tau bound to microtubules in tau_45–230-expressing hippocampal neurons when compared to controls. In addition, the presence of this tau fragment induced a transient increase in tyrosinated tubulin, a marker of unstable microtubules, followed by a significant decrease in the levels of this tubulin isoform. The data obtained also showed a significant reduction in actin filaments in tau_45–230-expressing neurons. These changes in microtubules and actin filaments correlated with delayed neurite elongation and axonal differentiation in the presence of this tau fragment. Together, these results suggest that tau_45–230 could exert its toxic effects, at least in part, by modifying the composition of the neuronal cytoskeleton and impairing neurite elongation in neurons undergoing degeneration.
Carbon nanomaterials and amyloid-beta interactions: potentials for the detection and treatment of Alzheimer's disease?
2019, Pharmacological Research
Carbon-based nanomaterials have unique physicochemical properties relevant to the diagnosis and treatment of various diseases. There have been many reports indicating that carbon nanomaterials (CNMs) can interact and perturb biomolecules such as proteins and amyloid structures. This review is an attempt to reflect the role of CNMs in the treatment and detection of Alzheimer's disease (AD). The potential of CNMs in the field of neuroscience has also been reviewed. The exposure to CNMs ends up with effective radical and peptide scavenging structures, which, in turn, inhibit further formation and progression of amyloid fibrils. However, the effect of CNMs on initial nucleation and lag phase in this process may promote fibrillation. We have discussed the controversy that whether CNMs promote or inhibit the formation of amyloid beta (Aβ) fibrils to help preventing their toxicity or enhancing their therapeutic effects.
Tau<inf>45-230</inf> association with the cytoskeleton and membrane-bound organelles: Functional implications in neurodegeneration
2017, Neuroscience
Citation Excerpt :
It is worth noting that no tau45-230 dimers were identified in hippocampal neurons treated with Aβ. The presence of these dimers could be masked due to their similar molecular weight to those of the adult full-length tau isoforms expressed in mature hippocampal neurons (Ferreira et al., 1997). Alternatively, the lack of tau45-230 dimerization in Aβ-treated neurons could reflect a differentially regulated process depending on specific stages of development (young vs. mature neurons) or levels of tau45-230 accumulated under these experimental conditions.
The dysregulation of posttranslational modifications of the microtubule-associated protein (MAP) tau plays a key role in Alzheimer’s disease (AD) and related disorders. Thus, we have previously shown that beta amyloid (Aβ)-induced neurotoxicity was mediated, at least in part, by tau cleavage into the tau_45-230 fragment. However, the mechanisms underlying the toxicity of tau_45-230 remain unknown. To get insights into such mechanisms, we first determined the subcellular localization of this tau fragment in hippocampal neurons. Tau_45-230 was easily detectable in cell bodies and processes extended by these neurons. In addition, cell extraction experiments performed using Triton X-100 and saponin showed that a pool of tau_45-230 was associated with the cytoskeleton and the cytoskeleton plus membrane-bound organelles, respectively, in cultured hippocampal neurons. Furthermore, they suggested that these associations were independent of the presence of full-length tau. We also assessed whether this tau fragment could alter axonal transport. Our results indicated that tau_45-230 significantly reduced the number of organelles transported along hippocampal axons. This altered axonal transport did not correlate with changes in the total number of organelles present in these cells or in motor protein levels. Together these results suggested that tau_45-230 could exert its toxic effects by partially blocking axonal transport along microtubules thus contributing to the early pathology of AD.
Characterization of neuronal tau protein as a target of extracellular signal-regulated kinase
2016, Journal of Biological Chemistry
Tau neuronal protein has a central role in neurodegeneration and is implicated in Alzheimer disease development. Abnormal phosphorylation of Tau impairs its interaction with other proteins and is associated with its dysregulation in pathological conditions. Molecular mechanisms leading to hyperphosphorylation of Tau in pathological conditions are unknown. Here, we characterize phosphorylation of Tau by extracellular-regulated kinase (ERK2), a mitogen-activated kinase (MAPK) that responds to extracellular signals. Analysis of in vitro phosphorylated Tau by activated recombinant ERK2 with nuclear magnetic resonance spectroscopy (NMR) reveals phosphorylation of 15 Ser/Thr sites. In vitro phosphorylation of Tau using rat brain extract and subsequent NMR analysis identifies the same sites. Phosphorylation with rat brain extract is known to transform Tau into an Alzheimer disease-like state. Our results indicate that phosphorylation of Tau by ERK2 alone is sufficient to produce the same characteristics. We further investigate the mechanism of ERK2 phosphorylation of Tau. Kinases are known to recognize their protein substrates not only by their specificity for a targeted Ser or Thr phosphorylation site but also by binding to linear-peptide motifs called docking sites. We identify two main ERK2 docking sites in Tau sequence using NMR. Our results suggest that ERK2 dysregulation in Alzheimer disease could lead to abnormal phosphorylation of Tau resulting in the pathology of the disease.

View all citing articles on Scopus

^☆: G. A. BankerK. Goslin, Eds.

¹: To whom correspondence should be addressed at present address: Department of Cell and Molecular Biology and Institute for Neuroscience, Northwestern University, Searle Building Room 5-474, 320 East Superior Street, Chicago, IL 60611. Fax: (312) 503 7345. E-mail: [email protected].

View full text

Regular ArticleSelective Phosphorylation of Adult Tau Isoforms in Mature Hippocampal Neurons Exposed to Fibrillar Aβ☆

Abstract

Neuron

Neuron

J. Biol. Chem.

Dev. Brain Res.

Neuron

FEBS Lett.

Neurosci. Lett.

FEBS Lett.

Neuron

Neuron

FEBS Lett.

J. Biol. Chem.

FEBS Lett.

Neuron

Mol. Brain Res.

J. Biol. Chem.

FEBS Lett.

FEBS Lett.

Cell

J. Biol. Chem.

Anal. Biochem.

Tau protein kinase II is involved in the regulation of the normal phosphorylation state of tau protein

J. Neurochem.

Abnormal Alzheimer's like phosphorylation of tau protein by cyclin-dependent cdk2 and cdk5

FEBS Lett.

Assay of protein in the presence of interfering material

Anal. Biochem.

Growth of a rat neuroblastoma cell line in serum-free supplemented media

Proc. Natl. Acad. Sci. USA

Developmental changes in τ phosphorylation: Foetal τ is transiently phosphorylated in a manner similar to paired helical filament τ characteristic of Alzheimer's disease

J. Neurochem.

Inhibition of neuronal polarity by tau antisense oligonucleotides in primary cerebellar neurones

Nature

Regular Article
Selective Phosphorylation of Adult Tau Isoforms in Mature Hippocampal Neurons Exposed to Fibrillar Aβ☆