Assembly of tau in transgenic animals expressing P301L tau

Alteration of phosphorylation and solubility

Naruhiko Sahara, Jada Lewis, Michael Deture, Eileen McGowan, Dennis W Dickson, Mike Hutton, Shu Hui Yen

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Transgenic mice (JNPL3), which develop neurofibrillary degeneration and express four-repeat human tau with P301L missense mutation, were characterized biochemically to determine whether the development of aggregated tau from soluble tau involves an intermediate stage. Homogenates from mice of different ages were separated into buffer-soluble (S1), sarkosyl- and salt-extractable (S2) and sarkosyl-insoluble pellet (P3) fractions, and analyzed for human tau distribution, phosphorylation and filament formation. S1 and S2 fractions contained 50-60-kDa tau whereas the S2 fraction also had 64-kDa tau. The level of tau in the P3 fraction increased in an age-dependent manner and correlated positively with the soluble tau concentration. The P3 fraction from 2.5-6.5-month-old mice contained 64- and 50-60-kDa tau, whereas that from 8.5-month and older transgenic animals contained mostly 64-kDa and higher molecular weight tau. The S2 and P3 fractions contained comparable amounts of 64-kDa tau. The 64-kDa tau was predominantly human, and phosphorylated at multiple sites: Thr181, Ser202/Thr205, Thr212, Thr231, Ser262, Ser396/Ser404, Ser409 and Ser422. Most of these sites were phosphorylated to a lesser extent in S2 than in P3 fractions. Tau polymers were detected in P3 fractions from 3-month and older female JNPL3 mice, but not in non-transgenic controls. The results suggest that tau in S2 represents an intermediate from which insoluble tau is derived, and that phosphorylation may play a role in filament formation and/or stabilization.

Original languageEnglish (US)
Pages (from-to)1498-1508
Number of pages11
JournalJournal of Neurochemistry
Volume83
Issue number6
DOIs
StatePublished - Dec 2002

Fingerprint

Genetically Modified Animals
Phosphorylation
Solubility
Animals
Buffers
Polymers
Stabilization
Salts
Molecular weight
Missense Mutation
Transgenic Mice
Molecular Weight
sarkosyl

Keywords

  • P301L tau
  • Phosphorylation
  • Solubility
  • Transgenic

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Assembly of tau in transgenic animals expressing P301L tau : Alteration of phosphorylation and solubility. / Sahara, Naruhiko; Lewis, Jada; Deture, Michael; McGowan, Eileen; Dickson, Dennis W; Hutton, Mike; Yen, Shu Hui.

In: Journal of Neurochemistry, Vol. 83, No. 6, 12.2002, p. 1498-1508.

Research output: Contribution to journalArticle

Sahara, Naruhiko ; Lewis, Jada ; Deture, Michael ; McGowan, Eileen ; Dickson, Dennis W ; Hutton, Mike ; Yen, Shu Hui. / Assembly of tau in transgenic animals expressing P301L tau : Alteration of phosphorylation and solubility. In: Journal of Neurochemistry. 2002 ; Vol. 83, No. 6. pp. 1498-1508.
@article{422c34597460443bb8f34902a5e7bb67,
title = "Assembly of tau in transgenic animals expressing P301L tau: Alteration of phosphorylation and solubility",
abstract = "Transgenic mice (JNPL3), which develop neurofibrillary degeneration and express four-repeat human tau with P301L missense mutation, were characterized biochemically to determine whether the development of aggregated tau from soluble tau involves an intermediate stage. Homogenates from mice of different ages were separated into buffer-soluble (S1), sarkosyl- and salt-extractable (S2) and sarkosyl-insoluble pellet (P3) fractions, and analyzed for human tau distribution, phosphorylation and filament formation. S1 and S2 fractions contained 50-60-kDa tau whereas the S2 fraction also had 64-kDa tau. The level of tau in the P3 fraction increased in an age-dependent manner and correlated positively with the soluble tau concentration. The P3 fraction from 2.5-6.5-month-old mice contained 64- and 50-60-kDa tau, whereas that from 8.5-month and older transgenic animals contained mostly 64-kDa and higher molecular weight tau. The S2 and P3 fractions contained comparable amounts of 64-kDa tau. The 64-kDa tau was predominantly human, and phosphorylated at multiple sites: Thr181, Ser202/Thr205, Thr212, Thr231, Ser262, Ser396/Ser404, Ser409 and Ser422. Most of these sites were phosphorylated to a lesser extent in S2 than in P3 fractions. Tau polymers were detected in P3 fractions from 3-month and older female JNPL3 mice, but not in non-transgenic controls. The results suggest that tau in S2 represents an intermediate from which insoluble tau is derived, and that phosphorylation may play a role in filament formation and/or stabilization.",
keywords = "P301L tau, Phosphorylation, Solubility, Transgenic",
author = "Naruhiko Sahara and Jada Lewis and Michael Deture and Eileen McGowan and Dickson, {Dennis W} and Mike Hutton and Yen, {Shu Hui}",
year = "2002",
month = "12",
doi = "10.1046/j.1471-4159.2002.01241.x",
language = "English (US)",
volume = "83",
pages = "1498--1508",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "6",

}

TY - JOUR

T1 - Assembly of tau in transgenic animals expressing P301L tau

T2 - Alteration of phosphorylation and solubility

AU - Sahara, Naruhiko

AU - Lewis, Jada

AU - Deture, Michael

AU - McGowan, Eileen

AU - Dickson, Dennis W

AU - Hutton, Mike

AU - Yen, Shu Hui

PY - 2002/12

Y1 - 2002/12

N2 - Transgenic mice (JNPL3), which develop neurofibrillary degeneration and express four-repeat human tau with P301L missense mutation, were characterized biochemically to determine whether the development of aggregated tau from soluble tau involves an intermediate stage. Homogenates from mice of different ages were separated into buffer-soluble (S1), sarkosyl- and salt-extractable (S2) and sarkosyl-insoluble pellet (P3) fractions, and analyzed for human tau distribution, phosphorylation and filament formation. S1 and S2 fractions contained 50-60-kDa tau whereas the S2 fraction also had 64-kDa tau. The level of tau in the P3 fraction increased in an age-dependent manner and correlated positively with the soluble tau concentration. The P3 fraction from 2.5-6.5-month-old mice contained 64- and 50-60-kDa tau, whereas that from 8.5-month and older transgenic animals contained mostly 64-kDa and higher molecular weight tau. The S2 and P3 fractions contained comparable amounts of 64-kDa tau. The 64-kDa tau was predominantly human, and phosphorylated at multiple sites: Thr181, Ser202/Thr205, Thr212, Thr231, Ser262, Ser396/Ser404, Ser409 and Ser422. Most of these sites were phosphorylated to a lesser extent in S2 than in P3 fractions. Tau polymers were detected in P3 fractions from 3-month and older female JNPL3 mice, but not in non-transgenic controls. The results suggest that tau in S2 represents an intermediate from which insoluble tau is derived, and that phosphorylation may play a role in filament formation and/or stabilization.

AB - Transgenic mice (JNPL3), which develop neurofibrillary degeneration and express four-repeat human tau with P301L missense mutation, were characterized biochemically to determine whether the development of aggregated tau from soluble tau involves an intermediate stage. Homogenates from mice of different ages were separated into buffer-soluble (S1), sarkosyl- and salt-extractable (S2) and sarkosyl-insoluble pellet (P3) fractions, and analyzed for human tau distribution, phosphorylation and filament formation. S1 and S2 fractions contained 50-60-kDa tau whereas the S2 fraction also had 64-kDa tau. The level of tau in the P3 fraction increased in an age-dependent manner and correlated positively with the soluble tau concentration. The P3 fraction from 2.5-6.5-month-old mice contained 64- and 50-60-kDa tau, whereas that from 8.5-month and older transgenic animals contained mostly 64-kDa and higher molecular weight tau. The S2 and P3 fractions contained comparable amounts of 64-kDa tau. The 64-kDa tau was predominantly human, and phosphorylated at multiple sites: Thr181, Ser202/Thr205, Thr212, Thr231, Ser262, Ser396/Ser404, Ser409 and Ser422. Most of these sites were phosphorylated to a lesser extent in S2 than in P3 fractions. Tau polymers were detected in P3 fractions from 3-month and older female JNPL3 mice, but not in non-transgenic controls. The results suggest that tau in S2 represents an intermediate from which insoluble tau is derived, and that phosphorylation may play a role in filament formation and/or stabilization.

KW - P301L tau

KW - Phosphorylation

KW - Solubility

KW - Transgenic

UR - http://www.scopus.com/inward/record.url?scp=0036904519&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036904519&partnerID=8YFLogxK

U2 - 10.1046/j.1471-4159.2002.01241.x

DO - 10.1046/j.1471-4159.2002.01241.x

M3 - Article

VL - 83

SP - 1498

EP - 1508

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 6

ER -