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Cytoplasmic accumulation of TDP-43 in circulating lymphomonocytes of ALS patients with and without TARDBP mutations

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Abstract

TDP-43, encoded by TARDBP, is a ubiquitously expressed, primarily nuclear protein. In recent years, TDP-43 has been identified as the major pathological protein in ALS due to its mislocalisation in the cytoplasm of motor neurons of patients with and without TARDBP mutations and expression in forms that do not match its predicted molecular weight. In this study, the TDP-43 profile was investigated using western immunoblot analysis in whole lysates, nuclei and cytoplasm of circulating lymphomonocytes from 16 ALS patients, 4 with (ALS/TDP+) and 12 without (ALS/TDP−) TARDBP mutations in the protein C-terminal domain, and thirteen age-matched, healthy donors (controls). Three disease-unaffected first-degree relatives of an ALS/TDP+ patient were also included: one carried the parent mutation (Rel/TDP+) whereas the other two did not (Rel/TDP−). In all ALS patients, relatives and controls, TDP-43 retained the predicted molecular weight in whole cell lysates and nuclei, but in the cytoplasm its molecular weight was slightly smaller than expected. In quantitative terms, TDP-43 was expressed at approximately the same levels in whole cell lysates of ALS patients, relatives and controls. In contrast, TDP-43 accumulated in the cytoplasm with concomitant nuclear depletion in all ALS/TDP+ patients, in about 50% of ALS/TDP− patients and in the Rel/TDP+ subject compared to the controls. In the remaining ALS/TDP− patients and in the two Rel/TDP− subjects, TDP-43 matched the control levels in both subcellular compartments. Were these findings further confirmed, circulating lymphomonocytes could be informative of TDP-43 mislocalisation in nervous tissue and used as a biomarker for future disease risk.

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Acknowledgments

Supported by Compagnia di San Paolo, Turin (Italy), Grant No. 2004.1424, and Regione Piemonte (Italy), Grant No. 204.2009.

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Authors and Affiliations

  1. Department of Medicine and Experimental Oncology, Section of Biochemistry, University of Turin, Via Michelangelo n 27b, 10126, Turin, Italy

    Giovanni De Marco, Elisa Lupino, Barbara Buccinnà, Cristina Ramondetti, Annarosa Lomartire, Maria Teresa Rinaudo & Marco Piccinini

  2. Department of Neuroscience, University of Turin, Via Cherasco n 15, 10126, Turin, Italy

    Giovanni De Marco, Andrea Calvo, Cristina Moglia & Silvia Grifoni

  3. Department of Neuroscience, University of Turin and Neuroscience Institute of Turin (NIT), Via Cherasco n 15, 10126, Turin, Italy

    Maria Teresa Giordana & Adriano Chiò

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  1. Giovanni De Marco
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Correspondence to Maria Teresa Rinaudo.

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De Marco, G., Lupino, E., Calvo, A. et al. Cytoplasmic accumulation of TDP-43 in circulating lymphomonocytes of ALS patients with and without TARDBP mutations. Acta Neuropathol 121, 611–622 (2011). https://doi.org/10.1007/s00401-010-0786-7

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  • DOI: https://doi.org/10.1007/s00401-010-0786-7

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