Acta Neuropathologica

, Volume 117, Issue 2, pp 137–149

Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies

  • Manuela Neumann
  • Linda K. Kwong
  • Edward B. Lee
  • Elisabeth Kremmer
  • Andrew Flatley
  • Yan Xu
  • Mark S. Forman
  • Dirk Troost
  • Hans A. Kretzschmar
  • John Q. Trojanowski
  • Virginia M.-Y. Lee
Original Paper

DOI: 10.1007/s00401-008-0477-9

Cite this article as:
Neumann, M., Kwong, L.K., Lee, E.B. et al. Acta Neuropathol (2009) 117: 137. doi:10.1007/s00401-008-0477-9

Abstract

Accumulation of hyperphosphorylated, ubiquitinated and N-terminally truncated TAR DNA-binding protein (TDP-43) is the pathological hallmark lesion in most familial and sporadic forms of FTLD-U and ALS, which can be subsumed as TDP-43 proteinopathies. In order to get more insight into the role of abnormal phosphorylation in the disease process, the identification of specific phosphorylation sites and the generation of phosphorylation-specific antibodies are mandatory. Here, we developed and characterized novel rat monoclonal antibodies (1D3 and 7A9) raised against phosphorylated S409/410 of TDP-43. These antibodies were used to study the presence of S409/410 phosphorylation by immunohistochemistry and biochemical analysis in a large series of 64 FTLD-U cases with or without motor neuron disease including familial cases with mutations in progranulin (n = 5), valosin-containing protein (n = 4) and linkage to chromosome 9p (n = 4), 18 ALS cases as well as other neurodegenerative diseases with concomitant TDP-43 pathology (n = 5). Our data demonstrate that phosphorylation of S409/410 of TDP-43 is a highly consistent feature in pathologic inclusions in the whole spectrum of sporadic and familial forms of TDP-43 proteinopathies. Physiological nuclear TDP-43 was not detectable with these mAbs by immunohistochemistry and by immunoblot analyses. While the accumulation of phosphorylated C-terminal fragments was a robust finding in the cortical brain regions of FTLD-U and ALS, usually being much more abundant than the phosphorylated full-length TDP-43 band, spinal cord samples revealed a predominance of full-length TDP-43 over C-terminal fragments. This argues for a distinct TDP-43 species composition in inclusions in cortical versus spinal cord cells. Overall, these mAbs are powerful tools for the highly specific detection of disease-associated abnormal TDP-43 species and will be extremely useful for the neuropathological routine diagnostics of TDP-43 proteinopathies and for the investigation of emerging cellular and animal models for TDP-43 proteinopathies.

Supplementary material

401_2008_477_MOESM1_ESM.fdf (69 kb)
Additional phosphopeptides used for immunization (FDF 69 kb)

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Manuela Neumann
    • 1
    • 2
  • Linda K. Kwong
    • 3
  • Edward B. Lee
    • 3
  • Elisabeth Kremmer
    • 4
  • Andrew Flatley
    • 4
  • Yan Xu
    • 3
  • Mark S. Forman
    • 5
  • Dirk Troost
    • 6
  • Hans A. Kretzschmar
    • 2
  • John Q. Trojanowski
    • 3
  • Virginia M.-Y. Lee
    • 3
  1. 1.Institute of NeuropathologyUniversity Hospital of ZürichZurichSwitzerland
  2. 2.Center for Neuropathology and Prion ResearchLudwig-Maximilians UniversityMunichGermany
  3. 3.Department of Pathology and Laboratory Medicine, Center for Neurodegenerative Disease Research, Institute on AgingUniversity of Pennsylvania School of MedicinePhiladelphiaUSA
  4. 4.Helmholtz Center Munich, German Research Center for Environmental Health (GmbH)Institute of Molecular ImmunologyMunichGermany
  5. 5.Department of Experimental MedicineMerck Research LaboratoriesNorth WalesUSA
  6. 6.Department of Neuropathology, Academic Medical CentreUniversity of AmsterdamAmsterdamThe Netherlands