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Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis

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Abstract

Abnormal TDP-43 aggregation is a prominent feature in the neuropathology of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration. Mutations in TARDBP, the gene encoding TDP-43, cause some cases of ALS. The normal function of TDP-43 remains incompletely understood. To better understand TDP-43 biology, we generated mutant mice carrying a genetrap disruption of Tardbp. Mice homozygous for loss of TDP-43 are not viable. TDP-43 deficient embryos die about day 7.5 of embryonic development thereby demonstrating that TDP-43 protein is essential for normal prenatal development and survival. However, heterozygous Tardbp mutant mice exhibit signs of motor disturbance and muscle weakness. Compared with wild type control littermates, Tardbp +/− animals have significantly decreased forelimb grip strength and display deficits in a standard inverted grid test despite no evidence of pathologic changes in motor neurons. Thus, TDP-43 is essential for viability, and mild reduction in TDP-43 function is sufficient to cause motor deficits without degeneration of motor neurons.

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References

  1. Abhyankar MM, Urekar C, Reddi PP (2007) A novel CpG-free vertebrate insulator silences the testis-specific SP-10 gene in somatic tissues: role for TDP-43 in insulator function. J Biol Chem 282:36143–36154

    Article  CAS  PubMed  Google Scholar 

  2. Amador-Ortiz C, Lin WL, Ahmed Z, Personett D, Davies P, Duara R, Graff-Radford NR, Hutton ML, Dickson DW (2007) TDP-43 immunoreactivity in hippocampal sclerosis and Alzheimer’s disease. Ann Neurol 61:435–445

    Article  CAS  PubMed  Google Scholar 

  3. Aubin JE, Liu F, Candeliere GA (2002) Single-cell PCR methods for studying stem cells and progenitors. Methods Mol Biol 185:403–415

    CAS  PubMed  Google Scholar 

  4. Ayala YM, Misteli T, Baralle FE (2008) TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression. Proc Natl Acad Sci USA 105:3785–3789

    Article  CAS  PubMed  Google Scholar 

  5. Ayala YM, Pantano S, D’Ambrogio A, Buratti E, Brindisi A, Marchetti C, Romano M, Baralle FE (2005) Human, Drosophila, and C. elegans TDP43: nucleic acid binding properties and splicing regulatory function. J Mol Biol 348:575–588

    Article  CAS  PubMed  Google Scholar 

  6. Benajiba L, Le Ber I, Camuzat A, Lacoste M, Thomas-Anterion C, Couratier P, Legallic S, Salachas F, Hannequin D, Decousus M, Lacomblez L, Guedj E, Golfier V, Camu W, Dubois B, Campion D, Meininger V, Brice A (2009) TARDBP mutations in motoneuron disease with frontotemporal lobar degeneration. Ann Neurol 65:470–473

    Article  CAS  PubMed  Google Scholar 

  7. Bose JK, Wang IF, Hung L, Tarn WY, Shen CK (2008) TDP-43 overexpression enhances exon 7 inclusion during the survival of motor neuron pre-mRNA splicing. J Biol Chem 283:28852–28859

    Article  CAS  PubMed  Google Scholar 

  8. Buratti E, Baralle FE (2008) Multiple roles of TDP-43 in gene expression, splicing regulation, and human disease. Front Biosci 13:867–878

    Article  CAS  PubMed  Google Scholar 

  9. Buratti E, Dork T, Zuccato E, Pagani F, Romano M, Baralle FE (2001) Nuclear factor TDP-43 and SR proteins promote in vitro and in vivo CFTR exon 9 skipping. EMBO J 20:1774–1784

    Article  CAS  PubMed  Google Scholar 

  10. Butchbach ME, Edwards JD, Burghes AH (2007) Abnormal motor phenotype in the SMNDelta7 mouse model of spinal muscular atrophy. Neurobiol Dis 27:207–219

    Article  CAS  PubMed  Google Scholar 

  11. D’Souza I, Poorkaj P, Hong M, Nochlin D, Lee VM-Y, Bird TD, Schellenberg GD (1999) Missense and silent tau gene mutations cause front temporal dementia with parkinsonism—chromosome 17 type by affecting multiple alternative RNA splicing regulatory elements. Proc Natl Acad Sci USA 96:5598–5603

    Article  PubMed  Google Scholar 

  12. Daoud H, Valdmanis PN, Kabashi E, Dion P, Dupre N, Camu W, Meininger V, Rouleau GA (2009) Contribution of TARDBP mutations to sporadic amyotrophic lateral sclerosis. J Med Genet 46:112–114

    Article  CAS  PubMed  Google Scholar 

  13. Feiguin F, Godena VK, Romano G, D’Ambrogio A, Klima R, Baralle FE (2009) Depletion of TDP-43 affects Drosophila motoneurons terminal synapsis and locomotive behavior. FEBS Lett 583:1586–1592

    Article  CAS  PubMed  Google Scholar 

  14. Forman MS, Lal D, Zhang B, Dabir DV, Swanson E, Lee VMY, Trojanowski JQ (2005) Transgenic mouse model of tau pathology in astrocytes leading to nervous system degeneration. J Neurosci 25:3539–3550

    Article  CAS  PubMed  Google Scholar 

  15. Geser F, Martinez-Lage M, Kwong LK, Lee VM, Trojanowski JQ (2009) Amyotrophic lateral sclerosis, frontotemporal dementia and beyond: the TDP-43 diseases. J Neurol 256(8):1205–1214

    Article  PubMed  Google Scholar 

  16. Gitcho MA, Baloh RH, Chakraverty S, Mayo K, Norton JB, Levitch D, Hatanpaa KJ, White CL III, Bigio EH, Caselli R, Baker M, Al-Lozi MT, Morris JC, Pestronk A, Rademakers R, Goate AM, Cairns NJ (2008) TDP-43 A315T mutation in familial motor neuron disease. Ann Neurol 63:535–538

    Article  CAS  PubMed  Google Scholar 

  17. Glynn D, Drew CJ, Reim K, Brose N, Morton AJ (2005) Profound ataxia in complexin I knockout mice masks a complex phenotype that includes exploratory and habituation deficits. Hum Mol Genet 14:2369–2385

    Article  CAS  PubMed  Google Scholar 

  18. Goedert M, Jakes R, Crowther RA (1999) Effects of frontotemporal dementia FTDP-17 mutations on heparin-induced assembly of tau filaments. FEBS Lett 450:306–311

    Article  CAS  PubMed  Google Scholar 

  19. Hutton M, Lendon CL, Rizzu P, Baker M, Froelich S, Houlden H, Pickering-Brown S, Chakraverty S, Isaacs A, Grover A, Hackett J, Adamson J, Lincoln S, Dickson D, Davies P, Petersen RC, Stevens M, de Graaff E, Wauters E, van Baren J, Hillebrand M, Joosse M, Kwon JM, Nowotny P, Che LK, Norton J, Morris JC, Reed LA, Trojanowski J, Basun H, Lannfelt L, Neystat M, Fahn S, Dark F, Tannenberg T, Dodd PR, Hayward N, Kwok JBJ, Schofield PR, Andreadis A, Snowden J, Craufurd D, Neary D, Owen F, Oostra BA, Hardy J, Goate A, van Swieten J, Mann D, Lynch T, Heutink P (1998) Association of missense and 5′-splice-site mutations in tau with the inherited dementia FTDP-17. Nature 393:702–705

    Article  CAS  PubMed  Google Scholar 

  20. Igaz LM, Kwong LK, Xu Y, Truax AC, Uryu K, Neumann M, Clark CM, Elman LB, Miller BL, Grossman M, McCluskey LF, Trojanowski JQ, Lee VM (2008) Enrichment of C-terminal fragments in TAR DNA-binding protein-43 cytoplasmic inclusions in brain but not in spinal cord of frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Am J Pathol 173:182–194

    Article  CAS  PubMed  Google Scholar 

  21. Kabashi E, Valdmanis PN, Dion P, Spiegelman D, McConkey BJ, Vande Velde C, Bouchard JP, Lacomblez L, Pochigaeva K, Salachas F, Pradat PF, Camu W, Meininger V, Dupre N, Rouleau GA (2008) TARDBP mutations in individuals with sporadic and familial amyotrophic lateral sclerosis. Nat Genet 40:572–574

    Article  CAS  PubMed  Google Scholar 

  22. Kamada M, Maruyama H, Tanaka E, Morino H, Wate R, Ito H, Kusaka H, Kawano Y, Miki T, Nodera H, Izumi Y, Kaji R, Kawakami H (2009) Screening for TARDBP mutations in Japanese familial amyotrophic lateral sclerosis. J Neurol Sci 284(1–2):69–71

    Google Scholar 

  23. Kwong LK, Uryu K, Trojanowski JQ, Lee VM (2008) TDP-43 proteinopathies: neurodegenerative protein misfolding diseases without amyloidosis. Neurosignals 16:41–51

    Article  CAS  PubMed  Google Scholar 

  24. Lemmens R, Race V, Hersmus N, Matthijs G, Van Den Bosch L, Van Damme P, Dubois B, Boonen S, Goris A, Robberecht W (2009) TDP-43 M311V mutation in familial amyotrophic lateral sclerosis. J Neurol Neurosurg Psychiatry 80:354–355

    Article  CAS  PubMed  Google Scholar 

  25. Lu Y, Ferris J, Gao FB (2009) Frontotemporal dementia and amyotrophic lateral sclerosis-associated disease protein TDP-43 promotes dendritic branching. Mol Brain 2:30

    Article  PubMed  Google Scholar 

  26. Mackenzie IR, Bigio EH, Ince PG, Geser F, Neumann M, Cairns NJ, Kwong LK, Forman MS, Ravits J, Stewart H, Eisen A, McClusky L, Kretzschmar HA, Monoranu CM, Highley JR, Kirby J, Siddique T, Shaw PJ, Lee VM, Trojanowski JQ (2007) Pathological TDP-43 distinguishes sporadic amyotrophic lateral sclerosis from amyotrophic lateral sclerosis with SOD1 mutations. Ann Neurol 61:427–434

    Article  CAS  PubMed  Google Scholar 

  27. Mackenzie IR, Neumann M, Bigio EH, Cairns NJ, Alafuzoff I, Kril J, Kovacs GG, Ghetti B, Halliday G, Holm IE, Ince PG, Kamphorst W, Revesz T, Rozemuller AJ, Kumar-Singh S, Akiyama H, Baborie A, Spina S, Dickson DW, Trojanowski JQ, Mann DM (2009) Nomenclature for neuropathologic subtypes of frontotemporal lobar degeneration: consensus recommendations. Acta Neuropathol 117:15–18

    Article  PubMed  Google Scholar 

  28. Mercado PA, Ayala YM, Romano M, Buratti E, Baralle FE (2005) Depletion of TDP 43 overrides the need for exonic and intronic splicing enhancers in the human apoA-II gene. Nucleic Acids Res 33:6000–6010

    Article  CAS  PubMed  Google Scholar 

  29. Nacharaju P, Lewis J, Easson C, Yen S, Hackett J, Hutton M, Yen SH (1999) Accelerated filament formation from tau protein with specific FTDP-17 missense mutations. FEBS Lett 447:195–199

    Article  CAS  PubMed  Google Scholar 

  30. Nakashima-Yasuda H, Uryu K, Robinson J, Xie SX, Hurtig H, Duda JE, Arnold SE, Siderowf A, Grossman M, Leverenz JB, Woltjer R, Lopez OL, Hamilton R, Tsuang DW, Galasko D, Masliah E, Kaye J, Clark CM, Montine TJ, Lee VM, Trojanowski JQ (2007) Co-morbidity of TDP-43 proteinopathy in Lewy body related diseases. Acta Neuropathol 114:221–229

    Article  CAS  PubMed  Google Scholar 

  31. Neumann M, Kwong LK, Lee EB, Kremmer E, Flatley A, Xu Y, Forman MS, Troost D, Kretzschmar HA, Trojanowski JQ, Lee VM (2009) Phosphorylation of S409/410 of TDP-43 is a consistent feature in all sporadic and familial forms of TDP-43 proteinopathies. Acta Neuropathol 117:137–149

    Article  CAS  PubMed  Google Scholar 

  32. Neumann M, Sampathu DM, Kwong LK, Truax AC, Micsenyi MC, Chou TT, Bruce J, Schuck T, Grossman M, Clark CM, McCluskey LF, Miller BL, Masliah E, Mackenzie IR, Feldman H, Feiden W, Kretzschmar HA, Trojanowski JQ, Lee VM (2006) Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science 314:130–133

    Article  CAS  PubMed  Google Scholar 

  33. Ou SH, Wu F, Harrich D, Garcia-Martinez LF, Gaynor RB (1995) Cloning and characterization of a novel cellular protein, TDP-43, that binds to human immunodeficiency virus type 1 TAR DNA sequence motifs. J Virol 69:3584–3596

    CAS  PubMed  Google Scholar 

  34. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning : a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  35. Sephton CF, Good SK, Atkin S, Dewey CM, Mayer P III, Herz J, Yu G (2010) TDP-43 is a developmentally regulated protein essential for early embryonic development. J Biol Chem 285(9):6826–6834

    Google Scholar 

  36. Shinzawa K, Sumi H, Ikawa M, Matsuoka Y, Okabe M, Sakoda S, Tsujimoto Y (2008) Neuroaxonal dystrophy caused by group VIA phospholipase A2 deficiency in mice: a model of human neurodegenerative disease. J Neurosci 28:2212–2220

    Article  CAS  PubMed  Google Scholar 

  37. Shu Z, Smith S, Wang L, Rice MC, Kmiec EB (1999) Disruption of muREC2/RAD51L1 in mice results in early embryonic lethality which can Be partially rescued in a p53(−/−) background. Mol Cell Biol 19:8686–8693

    CAS  PubMed  Google Scholar 

  38. Sreedharan J, Blair IP, Tripathi VB, Hu X, Vance C, Rogelj B, Ackerley S, Durnall JC, Williams KL, Buratti E, Baralle F, de Belleroche J, Mitchell JD, Leigh PN, Al-Chalabi A, Miller CC, Nicholson G, Shaw CE (2008) TDP-43 mutations in familial and sporadic amyotrophic lateral sclerosis. Science 319:1668–1672

    Article  CAS  PubMed  Google Scholar 

  39. Stack EC, Kubilus JK, Smith K, Cormier K, Del Signore SJ, Guelin E, Ryu H, Hersch SM, Ferrante RJ (2005) Chronology of behavioral symptoms and neuropathological sequela in R6/2 Huntington’s disease transgenic mice. J Comp Neurol 490:354–370

    Article  PubMed  Google Scholar 

  40. Stanford WL, Cohn JB, Cordes SP (2001) Gene-trap mutagenesis: past, present and beyond. Nat Rev Genet 2:756–768

    Article  CAS  PubMed  Google Scholar 

  41. Strong MJ, Volkening K, Hammond R, Yang W, Strong W, Leystra-Lantz C, Shoesmith C (2007) TDP43 is a human low molecular weight neurofilament (hNFL) mRNA-binding protein. Mol Cell Neurosci 35:320–327

    Article  CAS  PubMed  Google Scholar 

  42. Stryke D, Kawamoto M, Huang CC, Johns SJ, King LA, Harper CA, Meng EC, Lee RE, Yee A, L’Italien L, Chuang PT, Young SG, Skarnes WC, Babbitt PC, Ferrin TE (2003) BayGenomics: a resource of insertional mutations in mouse embryonic stem cells. Nucleic Acids Res 31:278–281

    Article  CAS  PubMed  Google Scholar 

  43. Tan CF, Eguchi H, Tagawa A, Onodera O, Iwasaki T, Tsujino A, Nishizawa M, Kakita A, Takahashi H (2007) TDP-43 immunoreactivity in neuronal inclusions in familial amyotrophic lateral sclerosis with or without SOD1 gene mutation. Acta Neuropathol 113:535–542

    Article  CAS  PubMed  Google Scholar 

  44. Tomiyama H, Kokubo Y, Sasaki R, Li Y, Imamichi Y, Funayama M, Mizuno Y, Hattori N, Kuzuhara S (2008) Mutation analyses in amyotrophic lateral sclerosis/parkinsonism-dementia complex of the Kii peninsula, Japan. Mov Disord 23:2344–2348

    Article  PubMed  Google Scholar 

  45. Van Deerlin VM, Leverenz JB, Bekris LM, Bird TD, Yuan W, Elman LB, Clay D, Wood EM, Chen-Plotkin AS, Martinez-Lage M, Steinbart E, McCluskey L, Grossman M, Neumann M, Wu IL, Yang WS, Kalb R, Galasko DR, Montine TJ, Trojanowski JQ, Lee VM, Schellenberg GD, Yu CE (2008) TARDBP mutations in amyotrophic lateral sclerosis with TDP-43 neuropathology: a genetic and histopathological analysis. Lancet Neurol 7:409–416

    Article  PubMed  Google Scholar 

  46. Wang HY, Wang IF, Bose J, Shen CK (2004) Structural diversity and functional implications of the eukaryotic TDP gene family. Genomics 83:130–139

    Article  CAS  PubMed  Google Scholar 

  47. Wang IF, Reddy NM, Shen CK (2002) Higher order arrangement of the eukaryotic nuclear bodies. Proc Natl Acad Sci USA 99:13583–13588

    Article  CAS  PubMed  Google Scholar 

  48. Weydt P, Hong SY, Kliot M, Moller T (2003) Assessing disease onset and progression in the SOD1 mouse model of ALS. Neuroreport 14:1051–1054

    Article  PubMed  Google Scholar 

  49. Winton MJ, Igaz LM, Wong MM, Kwong LK, Trojanowski JQ, Lee VM (2008) Disturbance of nuclear and cytoplasmic TAR DNA-binding protein (TDP-43) induces disease-like redistribution, sequestration, and aggregate formation. J Biol Chem 283(19):13302–13309

    Google Scholar 

  50. Wu LS, Cheng WC, Hou SC, Yan YT, Jiang ST, Shen CK (2010) TDP-43, a neuro-pathosignature factor, is essential for early mouse embryogenesis. Genesis 48:56–62

    CAS  PubMed  Google Scholar 

  51. Yokoseki A, Shiga A, Tan CF, Tagawa A, Kaneko H, Koyama A, Eguchi H, Tsujino A, Ikeuchi T, Kakita A, Okamoto K, Nishizawa M, Takahashi H, Onodera O (2008) TDP-43 mutation in familial amyotrophic lateral sclerosis. Ann Neurol 63:538–542

    Article  CAS  PubMed  Google Scholar 

  52. Yoshiyama Y, Higuchi M, Zhang B, Huang SM, Iwata N, Saido TC, Maeda J, Suhara T, Trojanowski JQ, Lee VM (2007) Synapse loss and microglial activation precede tangles in a P301S tauopathy mouse model. Neuron 53:337–351

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by NIH grant AG17586 to GDS, VMYL, and JQT. This work was also supported by grants to BCK from the Office of Research and Development Medical Research Service, Department of Veterans Affairs and NIH (NS064131). This work utilized facilities at the VA Puget Sound Health Care System, Seattle, Washington. We thank Eddie Lee for outstanding assistance examining neuropathology of Tardbp+/− animals and Elaine Loomis, Leojean Anderson, and Harmony Danner for excellent technical assistance. The lacZ monoclonal antibody was obtained from the Developmental Studies Hybridoma Bank maintained by the University of Iowa.

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Authors and Affiliations

  1. Geriatrics Research Education and Clinical Center, Veterans Affairs Puget Sound Health Care System, Seattle, WA, 98108, USA

    Brian C. Kraemer, Jeanna M. Wheeler & Linda C. Robinson

  2. Division of Gerontology and Geriatric Medicine, Department of Medicine, University of Washington, Seattle, WA, 98104, USA

    Brian C. Kraemer & Jeanna M. Wheeler

  3. Department of Pathology and Laboratory Medicine, Center for Neurodegenerative Disease Research and Institute on Aging, University of Pennsylvania School of Medicine, Philadelphia, PA, 19104-6100, USA

    Theresa Schuck, John Q. Trojanowski, Virginia M. Y. Lee & Gerard D. Schellenberg

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  1. Brian C. Kraemer
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Correspondence to Brian C. Kraemer.

Electronic supplementary material

401_2010_659_MOESM1_ESM.tif

Fig. S1 Analysis of genetrap targeted TDP-43 locus. Southern blotting was conducted as described (35) using random primed TDP-43 exon 1 and beta-geo coding sequence specific 32P-labeled probes and conditions as described for northern blotting. 20 micrograms of genomic DNA isolated from liver was digested with AflIII and PstI for TDP-43 and beta-geo specific probes respectively. Bands of ~3.6 kb corresponding to no genetrap and ~4.5 kb corresponding to a genetrap insertion in the TDP-43 locus were observed for the TDP-43 exon 1 probe. A single genetrap specific band was observed for the exon 1 probe corresponding to the 7.5 kb beta-geo containing genetrap inserted within the TDP-43 locus (TIFF 5,547 kb)

401_2010_659_MOESM2_ESM.tif

Fig. S2 Analysis of amino and carboxy terminal TDP-43 specific fragments inTardbp +/−mice. Immunoblotting of brain, heart, kidney, liver, lung, muscle, and spleentissue protein extracts. Duplicate blots were probed with a. N-terminal TDP-43 specificantibody. b. C-terminal specific TDP-43 antibody (TIFF 11,345 kb)

401_2010_659_MOESM3_ESM.tif

Fig. S3 Young Tardbp +/− animals do not display deficits in grip strength.a. Thirty day oldTardbp +/− animals (n = 16) hung on an inverted wire grid for the same lengthof time as control animals (n = 23) and b. also exhibit similar maximum forelimb gripstrength. Individual scores are the average of three trials, and the error bars representstandard deviation (TIFF 9,928 kb)

401_2010_659_MOESM4_ESM.tif

Fig. S4 Aged Tardbp +/− spinal cord is similar to wild type littermates.Shown here are images of sacral spinal cord of 28 month old mice. a. Tardbp +/+ and b.Tardbp +/− . As described in the text, there are no differences in cord these tissues betweenTardbp +/− and wild type mice. The bar in each panel = 100 um (TIFF 53,387 kb)

401_2010_659_MOESM5_ESM.tif

Fig. S5 Aged Tardbp +/− muscle is similar to wild type littermates.Shown here are images of gastronemius muscle of 28 month old mice. a. Tardbp +/+ and b.Tardbp +/−. As described in the text, there are no differences in cord these tissues betweenthe Tardbp +/− and wild type mice. The bar in each panel = 100 um (TIFF 53,517 kb)

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Kraemer, B.C., Schuck, T., Wheeler, J.M. et al. Loss of murine TDP-43 disrupts motor function and plays an essential role in embryogenesis. Acta Neuropathol 119, 409–419 (2010). https://doi.org/10.1007/s00401-010-0659-0

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