Characteristics of TBS-extractable hyperphosphorylated tau species: Aggregation intermediates in rTg4510 mouse brain

Naruhiko Sahara, Michael Deture, Yan Ren, Abdul Shukkur Ebrahim, Dongcheul Kang, Joshua Knight, Christiane Volbracht, Jan Torleif Pedersen, Dennis W Dickson, Shu Hui Yen, Jada Lewis

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Conditional overexpression of four-repeat human tau containing the P301L missense mutation in the rTg4510 mouse model of tauopathy leads to progressive accumulation of neurofibrillary tangles and hyperphosphorylated, sarkosyl-insoluble tau species, which are biochemically comparable to abnormal tau characteristic of hereditary tauopathies termed FTDP-17. To fully understand the impact of tau species at different stages of self-assembly on neurodegeneration, we fractionated rTg4510 brain representing several stages of tauopathy to obtain TBS-extractable (S1), high salt/sarkosyl-extractable (S3), and sarkosyl-insoluble (P3) fractions. Under reducing condition, the S1 fraction was demonstrated by western blotting to contain both 50-60 kDa normally-sized and 64 kDa tau. Both are thermo-stable, but the 64 kDa tau showed a higher degree of phosphorylation. Under non-reducing condition, nearly all TBS-extractable 64 kDa tau were detected as ∼130 kDa species consistent with the size of dimer. Quantitative analysis showed ∼80 times more 64 kDa tau in S1 than P3 fraction. Immunoelectron microscopy revealed tau-positive granules/short filaments in S1 fraction. These structures displayed MC1 immunoreactivities indicative of conformational/pathological change of tau. MC1 immunoreactivity was detected by dot blotting in samples from 2.5 month-old mice, whereas Ab39 immunoreactivity indicative of late stages of tau assembly was detected only in P3 fraction. Quantitative analysis also demonstrated a significant inverse correlation between brain weight and 64 kDa tau, but the level of TBS-extractable 64 kDa tau reflects neurodegeneration better than that of sarkosyl-insoluble 64 kDa tau. Together, the findings suggest that TBS-extractable 64 kDa tau production is a potential target for therapeutic intervention of tauopathies.

Original languageEnglish (US)
Pages (from-to)249-263
Number of pages15
JournalJournal of Alzheimer's Disease
Volume33
Issue number1
DOIs
StatePublished - 2013

Fingerprint

Tauopathies
Brain
Frontotemporal Dementia
Neurofibrillary Tangles
Immunoelectron Microscopy
Missense Mutation
Salts
Western Blotting
Phosphorylation
Weights and Measures
sarkosyl

Keywords

  • Dimer
  • FTDP-17
  • hyperphosphorylation
  • tau protein
  • tauopathy
  • transgenic mice

ASJC Scopus subject areas

  • Psychiatry and Mental health
  • Geriatrics and Gerontology
  • Clinical Psychology

Cite this

Characteristics of TBS-extractable hyperphosphorylated tau species : Aggregation intermediates in rTg4510 mouse brain. / Sahara, Naruhiko; Deture, Michael; Ren, Yan; Ebrahim, Abdul Shukkur; Kang, Dongcheul; Knight, Joshua; Volbracht, Christiane; Pedersen, Jan Torleif; Dickson, Dennis W; Yen, Shu Hui; Lewis, Jada.

In: Journal of Alzheimer's Disease, Vol. 33, No. 1, 2013, p. 249-263.

Research output: Contribution to journalArticle

Sahara, N, Deture, M, Ren, Y, Ebrahim, AS, Kang, D, Knight, J, Volbracht, C, Pedersen, JT, Dickson, DW, Yen, SH & Lewis, J 2013, 'Characteristics of TBS-extractable hyperphosphorylated tau species: Aggregation intermediates in rTg4510 mouse brain', Journal of Alzheimer's Disease, vol. 33, no. 1, pp. 249-263. https://doi.org/10.3233/JAD-2012-121093
Sahara, Naruhiko ; Deture, Michael ; Ren, Yan ; Ebrahim, Abdul Shukkur ; Kang, Dongcheul ; Knight, Joshua ; Volbracht, Christiane ; Pedersen, Jan Torleif ; Dickson, Dennis W ; Yen, Shu Hui ; Lewis, Jada. / Characteristics of TBS-extractable hyperphosphorylated tau species : Aggregation intermediates in rTg4510 mouse brain. In: Journal of Alzheimer's Disease. 2013 ; Vol. 33, No. 1. pp. 249-263.
@article{140e8c05873b4c97ad337a6df192914e,
title = "Characteristics of TBS-extractable hyperphosphorylated tau species: Aggregation intermediates in rTg4510 mouse brain",
abstract = "Conditional overexpression of four-repeat human tau containing the P301L missense mutation in the rTg4510 mouse model of tauopathy leads to progressive accumulation of neurofibrillary tangles and hyperphosphorylated, sarkosyl-insoluble tau species, which are biochemically comparable to abnormal tau characteristic of hereditary tauopathies termed FTDP-17. To fully understand the impact of tau species at different stages of self-assembly on neurodegeneration, we fractionated rTg4510 brain representing several stages of tauopathy to obtain TBS-extractable (S1), high salt/sarkosyl-extractable (S3), and sarkosyl-insoluble (P3) fractions. Under reducing condition, the S1 fraction was demonstrated by western blotting to contain both 50-60 kDa normally-sized and 64 kDa tau. Both are thermo-stable, but the 64 kDa tau showed a higher degree of phosphorylation. Under non-reducing condition, nearly all TBS-extractable 64 kDa tau were detected as ∼130 kDa species consistent with the size of dimer. Quantitative analysis showed ∼80 times more 64 kDa tau in S1 than P3 fraction. Immunoelectron microscopy revealed tau-positive granules/short filaments in S1 fraction. These structures displayed MC1 immunoreactivities indicative of conformational/pathological change of tau. MC1 immunoreactivity was detected by dot blotting in samples from 2.5 month-old mice, whereas Ab39 immunoreactivity indicative of late stages of tau assembly was detected only in P3 fraction. Quantitative analysis also demonstrated a significant inverse correlation between brain weight and 64 kDa tau, but the level of TBS-extractable 64 kDa tau reflects neurodegeneration better than that of sarkosyl-insoluble 64 kDa tau. Together, the findings suggest that TBS-extractable 64 kDa tau production is a potential target for therapeutic intervention of tauopathies.",
keywords = "Dimer, FTDP-17, hyperphosphorylation, tau protein, tauopathy, transgenic mice",
author = "Naruhiko Sahara and Michael Deture and Yan Ren and Ebrahim, {Abdul Shukkur} and Dongcheul Kang and Joshua Knight and Christiane Volbracht and Pedersen, {Jan Torleif} and Dickson, {Dennis W} and Yen, {Shu Hui} and Jada Lewis",
year = "2013",
doi = "10.3233/JAD-2012-121093",
language = "English (US)",
volume = "33",
pages = "249--263",
journal = "Journal of Alzheimer's Disease",
issn = "1387-2877",
publisher = "IOS Press",
number = "1",

}

TY - JOUR

T1 - Characteristics of TBS-extractable hyperphosphorylated tau species

T2 - Aggregation intermediates in rTg4510 mouse brain

AU - Sahara, Naruhiko

AU - Deture, Michael

AU - Ren, Yan

AU - Ebrahim, Abdul Shukkur

AU - Kang, Dongcheul

AU - Knight, Joshua

AU - Volbracht, Christiane

AU - Pedersen, Jan Torleif

AU - Dickson, Dennis W

AU - Yen, Shu Hui

AU - Lewis, Jada

PY - 2013

Y1 - 2013

N2 - Conditional overexpression of four-repeat human tau containing the P301L missense mutation in the rTg4510 mouse model of tauopathy leads to progressive accumulation of neurofibrillary tangles and hyperphosphorylated, sarkosyl-insoluble tau species, which are biochemically comparable to abnormal tau characteristic of hereditary tauopathies termed FTDP-17. To fully understand the impact of tau species at different stages of self-assembly on neurodegeneration, we fractionated rTg4510 brain representing several stages of tauopathy to obtain TBS-extractable (S1), high salt/sarkosyl-extractable (S3), and sarkosyl-insoluble (P3) fractions. Under reducing condition, the S1 fraction was demonstrated by western blotting to contain both 50-60 kDa normally-sized and 64 kDa tau. Both are thermo-stable, but the 64 kDa tau showed a higher degree of phosphorylation. Under non-reducing condition, nearly all TBS-extractable 64 kDa tau were detected as ∼130 kDa species consistent with the size of dimer. Quantitative analysis showed ∼80 times more 64 kDa tau in S1 than P3 fraction. Immunoelectron microscopy revealed tau-positive granules/short filaments in S1 fraction. These structures displayed MC1 immunoreactivities indicative of conformational/pathological change of tau. MC1 immunoreactivity was detected by dot blotting in samples from 2.5 month-old mice, whereas Ab39 immunoreactivity indicative of late stages of tau assembly was detected only in P3 fraction. Quantitative analysis also demonstrated a significant inverse correlation between brain weight and 64 kDa tau, but the level of TBS-extractable 64 kDa tau reflects neurodegeneration better than that of sarkosyl-insoluble 64 kDa tau. Together, the findings suggest that TBS-extractable 64 kDa tau production is a potential target for therapeutic intervention of tauopathies.

AB - Conditional overexpression of four-repeat human tau containing the P301L missense mutation in the rTg4510 mouse model of tauopathy leads to progressive accumulation of neurofibrillary tangles and hyperphosphorylated, sarkosyl-insoluble tau species, which are biochemically comparable to abnormal tau characteristic of hereditary tauopathies termed FTDP-17. To fully understand the impact of tau species at different stages of self-assembly on neurodegeneration, we fractionated rTg4510 brain representing several stages of tauopathy to obtain TBS-extractable (S1), high salt/sarkosyl-extractable (S3), and sarkosyl-insoluble (P3) fractions. Under reducing condition, the S1 fraction was demonstrated by western blotting to contain both 50-60 kDa normally-sized and 64 kDa tau. Both are thermo-stable, but the 64 kDa tau showed a higher degree of phosphorylation. Under non-reducing condition, nearly all TBS-extractable 64 kDa tau were detected as ∼130 kDa species consistent with the size of dimer. Quantitative analysis showed ∼80 times more 64 kDa tau in S1 than P3 fraction. Immunoelectron microscopy revealed tau-positive granules/short filaments in S1 fraction. These structures displayed MC1 immunoreactivities indicative of conformational/pathological change of tau. MC1 immunoreactivity was detected by dot blotting in samples from 2.5 month-old mice, whereas Ab39 immunoreactivity indicative of late stages of tau assembly was detected only in P3 fraction. Quantitative analysis also demonstrated a significant inverse correlation between brain weight and 64 kDa tau, but the level of TBS-extractable 64 kDa tau reflects neurodegeneration better than that of sarkosyl-insoluble 64 kDa tau. Together, the findings suggest that TBS-extractable 64 kDa tau production is a potential target for therapeutic intervention of tauopathies.

KW - Dimer

KW - FTDP-17

KW - hyperphosphorylation

KW - tau protein

KW - tauopathy

KW - transgenic mice

UR - http://www.scopus.com/inward/record.url?scp=84872441246&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84872441246&partnerID=8YFLogxK

U2 - 10.3233/JAD-2012-121093

DO - 10.3233/JAD-2012-121093

M3 - Article

C2 - 22941973

AN - SCOPUS:84872441246

VL - 33

SP - 249

EP - 263

JO - Journal of Alzheimer's Disease

JF - Journal of Alzheimer's Disease

SN - 1387-2877

IS - 1

ER -