Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS

Abstract

The nuclear transactive response DNA-binding protein 43 (TDP-43) undergoes relocalization to the cytoplasm with formation of cytoplasmic deposits in neurons in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Pathogenic mutations in the TDP-43-encoding TARDBP gene in familial ALS as well as non-mutant human TDP-43 have been utilized to model FTD/ALS in cell culture and animals, including mice. Here, we report novel A315T mutant TDP-43 transgenic mice, iTDP-43A315T, with controlled neuronal over-expression. Constitutive expression of human TDP-43A315T resulted in pronounced early-onset and progressive neurodegeneration, which was associated with compromised motor performance, spatial memory and disinhibition. Muscle atrophy resulted in reduced grip strength. Cortical degeneration presented with pronounced astrocyte activation. Using differential protein extraction from iTDP-43A315T brains, we found cytoplasmic localization, fragmentation, phosphorylation and ubiquitination and insolubility of TDP-43. Surprisingly, suppression of human TDP-43A315T expression in mice with overt neurodegeneration for only 1 week was sufficient to significantly improve motor and behavioral deficits, and reduce astrogliosis. Our data suggest that functional deficits in iTDP-43A315T mice are at least in part a direct and transient effect of the presence of TDP-43A315T. Furthermore, it illustrates the compensatory capacity of compromised neurons once transgenic TDP-43 is removed, with implications for future treatments.

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Acknowledgments

The authors like to thank the staff of the Biological Resources Centre Wallace Wurth for animal care. We also thank Prof Virginia Lee for anti mouse TDP-43 antibodies. This work has been supported by the National Health & Medical Research Council (NHMRC; #1081916, #1020562), the NHMRC Forefront Program grant (#1037746) and the Australian Research Council (ARC; #DP1096674, #DP130102027), Motor Neuron Disease Australia and the University of New South Wales. Y.D.K. is an ARC DECRA fellow (DE130101591). G.M.H. is an NHMRC Senior Principal Research Fellow (#1079679). L.M.I. is an NHMRC Senior Research Fellow (#1003083).

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Correspondence to Lars M. Ittner.

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Y. D. Ke, A. van Hummel and C. H. Stevens contributed equally to this work.

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Pole test deficits in 3 month-old iTDP-43A315T mice. Vertical pole test: Both 3 month-old mThy1.2-tTA(6)/pTRE-TDP-43A315T(13) and mThy1.2-tTA(15)/pTRE-TDP-43A315T(13) mice required significantly more time to reach the bottom when placed at the top of the pole, as compared to single transgenic mThy1.2-tTA(15), mThy1.2-tTA(6) and pTRE-TDP-43A315T(13), and non-transgenic (non-tg) control mice (***, P < 0.001, n = 14 (non-tg), n = 14 (pTRE-TDP-43A315T(13)), n = 6 (pTRE-TDP-43A315T(6)), n = 9 (mThy1.2-tTA(15)), n = 7 (mThy1.2-tTA(6)/pTRE-TDP-43A315T(13)), n = 10 (mThy1.2-tTA(15)/pTRE-TDP-43A315T(13)))

No overt microglial activation in iTDP-43A315T mice. Numbers and appearance of IBA-1-positive (green) microglia was indistinguishable in the cortex and hippocampus of mThy1.2-tTA(6)/pTRE-TDP-43A315T(13) iTDP-43A315T and single transgenic pTRE-TDP-43A315T(13) control (ctr) mice at 4.5 months of age. Human TDP-43 (hTDP-43) staining showed transgene expression in iTDP-43A315T mice. Scale bars, 100 µm

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Ke, Y.D., van Hummel, A., Stevens, C.H. et al. Short-term suppression of A315T mutant human TDP-43 expression improves functional deficits in a novel inducible transgenic mouse model of FTLD-TDP and ALS. Acta Neuropathol 130, 661–678 (2015). https://doi.org/10.1007/s00401-015-1486-0

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Keywords

  • TDP-43
  • Frontotemporal lobar degeneration
  • Amyotrophic lateral sclerosis
  • Mouse model
  • Pathogenic mutation