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Acta Neuropathologica

, Volume 132, Issue 1, pp 111–126 | Cite as

ECEL1 mutation implicates impaired axonal arborization of motor nerves in the pathogenesis of distal arthrogryposis

  • Kenichi NagataEmail author
  • Sumiko Kiryu-Seo
  • Hiromi Tamada
  • Fumi Okuyama-Uchimura
  • Hiroshi KiyamaEmail author
  • Takaomi C. SaidoEmail author
Original Paper

Abstract

The membrane-bound metalloprotease endothelin-converting enzyme-like 1 (ECEL1) has been newly identified as a causal gene of a specific type of distal arthrogryposis (DA). In contrast to most causal genes of DA, ECEL1 is predominantly expressed in neuronal cells, suggesting a unique neurogenic pathogenesis in a subset of DA patients with ECEL1 mutation. The present study analyzed developmental motor innervation and neuromuscular junction formation in limbs of the rodent homologue damage-induced neuronal endopeptidase (DINE)-deficient mouse. Whole-mount immunostaining was performed in DINE-deficient limbs expressing motoneuron-specific GFP to visualize motor innervation throughout the limb. Although DINE-deficient motor nerves displayed normal trajectory patterns from the spinal cord to skeletal muscles, they indicated impaired axonal arborization in skeletal muscles in the forelimbs and hindlimbs. Systematic examination of motor innervation in over 10 different hindlimb muscles provided evidence that DINE gene disruption leads to insufficient arborization of motor nerves after arriving at the skeletal muscle. Interestingly, the axonal arborization defect in foot muscles appeared more severe than in other hindlimb muscles, which was partially consistent with the proximal–distal phenotypic discordance observed in DA patients. Additionally, the number of innervated neuromuscular junction was significantly reduced in the severely affected DINE-deficient muscle. Furthermore, we generated a DINE knock-in (KI) mouse model with a pathogenic mutation, which was recently identified in DA patients. Axonal arborization defects were clearly detected in motor nerves of the DINE KI limb, which was identical to the DINE-deficient limb. Given that the encoded sequences, as well as ECEL1 and DINE expression profiles, are highly conserved between mouse and human, abnormal arborization of motor axons and subsequent failure of NMJ formation could be a primary cause of DA with ECEL1 mutation.

Keywords

Distal arthrogryposis DINE ECEL1 Motor nerve Axonal arborization Neuromuscular junction 

Notes

Acknowledgments

We thank Yukiko Nagai for secretarial assistance, Tomohiro Tanaka for his kind technical advice, Artur Kania and Kania lab members (especially Daniel Morales and Chris Law) for thoughtful discussion, Division for Medical Research Engineering, Nagoya University Graduate School of Medicine for the usage of transmission electron microscope, and Saido lab members for assistance. We are grateful to RIKEN BSI-Olympus Collaboration Center for imaging equipment and software, and Tetsuya Tajima and Kaori Higuchi from the center for technical support. Injection of the CRISPR/Cas9 system into mouse zygotes was performed at the RIKEN BSI-Research Resource Center. This work was financially supported by a Grant-in-Aid for Japan Society for the Promotion of Science Fellows; Japan Society for the Promotion of Science KAKENHI Grant No. 26860141; Special Postdoctoral Researchers Program in RIKEN; RIKEN Brain Science Institute.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

401_2016_1554_MOESM1_ESM.pdf (4.4 mb)
Supplementary material 1 (PDF 4487 kb)

References

  1. 1.
    Bamshad M, Jorde LB, Carey JC (1996) A revised and extended classification of the distal arthrogryposes. Am J Med Genet 65:277–281CrossRefPubMedGoogle Scholar
  2. 2.
    Bamshad M, Van Heest AE, Pleasure D (2009) Arthrogryposis: a review and update. J Bone Joint Surg Am 91(Suppl 4):40–46CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Barnett CP, Todd EJ, Ong R, Davis MR, Atkinson V, Allcock R, Laing N, Ravenscroft G (2014) Distal arthrogryposis type 5D with novel clinical features and compound heterozygous mutations in ECEL1. Am J Med Genet A 164:1846–1849CrossRefGoogle Scholar
  4. 4.
    Beals RK, Weleber RG (2004) Distal arthrogryposis 5: a dominant syndrome of peripheral contractures and ophthalmoplegia. Am J Med Genet A 131:67–70CrossRefPubMedGoogle Scholar
  5. 5.
    Benoit A, Vargas MA, Desgroseillers L, Boileau G (2004) Endothelin-converting enzyme-like 1 (ECEL1) is present both in the plasma membrane and in the endoplasmic reticulum. Biochem J 380:881–888. doi: 10.1042/bj20040215 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Burgess RW, Jucius TJ, Ackerman SL (2006) Motor axon guidance of the mammalian trochlear and phrenic nerves: dependence on the netrin receptor Unc5c and modifier loci. J Neurosci 26:5756–5766CrossRefPubMedGoogle Scholar
  7. 7.
    Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA et al (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339:819–823. doi: 10.1126/science.1231143 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    De Marco Garcia NV, Jessell TM (2008) Early motor neuron pool identity and muscle nerve trajectory defined by postmitotic restrictions in Nkx6.1 activity. Neuron 57:217–231CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Dieterich K, Quijano-Roy S, Monnier N, Zhou J, Faure J, Smirnow DA, Carlier R, Laroche C, Marcorelles P, Mercier S et al (2013) The neuronal endopeptidase ECEL1 is associated with a distinct form of recessive distal arthrogryposis. Hum Mol Genet 22:1483–1492CrossRefPubMedGoogle Scholar
  10. 10.
    Gurnett CA, Desruisseau DM, McCall K, Choi R, Meyer ZI, Talerico M, Miller SE, Ju JS, Pestronk A, Connolly AM et al (2010) Myosin binding protein C1: a novel gene for autosomal dominant distal arthrogryposis type 1. Hum Mol Genet 19:1165–1173CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Hall JG, Reed SD, Greene G (1982) The distal arthrogryposes: delineation of new entities–review and nosologic discussion. Am J Med Genet 11:185–239CrossRefPubMedGoogle Scholar
  12. 12.
    Huber AB, Kania A, Tran TS, Gu C, De Marco Garcia N, Lieberam I, Johnson D, Jessell TM, Ginty DD, Kolodkin AL (2005) Distinct roles for secreted semaphorin signaling in spinal motor axon guidance. Neuron 48:949–964CrossRefPubMedGoogle Scholar
  13. 13.
    Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821. doi: 10.1126/science.1225829 CrossRefPubMedGoogle Scholar
  14. 14.
    Kato R, Kiryu-Seo S, Kiyama H (2002) Damage-induced neuronal endopeptidase (DINE/ECEL) expression is regulated by leukemia inhibitory factor and deprivation of nerve growth factor in rat sensory ganglia after nerve injury. J Neurosci 22:9410–9418PubMedGoogle Scholar
  15. 15.
    Keane TM, Goodstadt L, Danecek P, White MA, Wong K, Yalcin B, Heger A, Agam A, Slater G, Goodson M et al (2011) Mouse genomic variation and its effect on phenotypes and gene regulation. Nature 477:289–294CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Khan AO, Shaheen R, Alkuraya FS (2014) The ECEL1-related strabismus phenotype is consistent with congenital cranial dysinnervation disorder. J Aapos 18:362–367CrossRefPubMedGoogle Scholar
  17. 17.
    Kiryu-Seo S, Sasaki M, Yokohama H, Nakagomi S, Hirayama T, Aoki S, Wada K, Kiyama H (2000) Damage-induced neuronal endopeptidase (DINE) is a unique metallopeptidase expressed in response to neuronal damage and activates superoxide scavengers. Proc Natl Acad Sci USA 97:4345–4350CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    MacLeod KJ, Fuller RS, Scholten JD, Ahn K (2001) Conserved cysteine and tryptophan residues of the endothelin-converting enzyme-1 CXAW motif are critical for protein maturation and enzyme activity. J Biol Chem 276:30608–30614. doi: 10.1074/jbc.M103928200 CrossRefPubMedGoogle Scholar
  19. 19.
    McMillin MJ, Below JE, Shively KM, Beck AE, Gildersleeve HI, Pinner J, Gogola GR, Hecht JT, Grange DK, Harris DJ et al (2013) Mutations in ECEL1 cause distal arthrogryposis type 5D. Am J Hum Genet 92:150–156CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Nagata K, Kiryu-Seo S, Kiyama H (2006) Localization and ontogeny of damage-induced neuronal endopeptidase mRNA-expressing neurons in the rat nervous system. Neuroscience 141:299–310CrossRefPubMedGoogle Scholar
  21. 21.
    Nagata K, Kiryu-Seo S, Maeda M, Yoshida K, Morita T, Kiyama H (2010) Damage-induced neuronal endopeptidase is critical for presynaptic formation of neuromuscular junctions. J Neurosci 30:6954–6962CrossRefPubMedGoogle Scholar
  22. 22.
    Ohba N, Kiryu-Seo S, Maeda M, Muraoka M, Ishii M, Kiyama H (2004) Expression of damage-induced neuronal endopeptidase (DINE) mRNA in peri-infarct cortical and thalamic neurons following middle cerebral artery occlusion. J Neurochem 91:956–964CrossRefPubMedGoogle Scholar
  23. 23.
    Patil SJ, Rai GK, Bhat V, Ramesh VA, Nagarajaram HA, Matalia J, Phadke SR (2014) Distal arthrogryposis type 5D with a novel ECEL1 gene mutation. Am J Med Genet A 164:2857–2862CrossRefGoogle Scholar
  24. 24.
    Pun S, Sigrist M, Santos AF, Ruegg MA, Sanes JR, Jessell TM, Arber S, Caroni P (2002) An intrinsic distinction in neuromuscular junction assembly and maintenance in different skeletal muscles. Neuron 34:357–370CrossRefPubMedGoogle Scholar
  25. 25.
    Qiu P, Shandilya H, D’Alessio JM, O’Connor K, Durocher J, Gerard GF (2004) Mutation detection using surveyor nuclease. Biotechniques 36:702–707PubMedGoogle Scholar
  26. 26.
    Schweizer A, Valdenaire O, Koster A, Lang Y, Schmitt G, Lenz B, Bluethmann H, Rohrer J (1999) Neonatal lethality in mice deficient in XCE, a novel member of the endothelin-converting enzyme and neutral endopeptidase family. J Biol Chem 274:20450–20456CrossRefPubMedGoogle Scholar
  27. 27.
    Shaaban S, Duzcan F, Yildirim C, Chan WM, Andrews C, Akarsu NA, Engle EC (2014) Expanding the phenotypic spectrum of ECEL1-related congenital contracture syndromes. Clin Genet 85:562–567CrossRefPubMedGoogle Scholar
  28. 28.
    Shaheen R, Al-Owain M, Khan AO, Zaki MS, Hossni HA, Al-Tassan R, Eyaid W, Alkuraya FS (2014) Identification of three novel ECEL1 mutations in three families with distal arthrogryposis type 5D. Clin Genet 85:568–572CrossRefPubMedGoogle Scholar
  29. 29.
    Sung SS, Brassington AM, Krakowiak PA, Carey JC, Jorde LB, Bamshad M (2003) Mutations in TNNT3 cause multiple congenital contractures: a second locus for distal arthrogryposis type 2B. Am J Hum Genet 73:212–214CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Toydemir RM, Rutherford A, Whitby FG, Jorde LB, Carey JC, Bamshad MJ (2006) Mutations in embryonic myosin heavy chain (MYH3) cause Freeman-Sheldon syndrome and Sheldon-Hall syndrome. Nat Genet 38:561–565CrossRefPubMedGoogle Scholar
  31. 31.
    Valdenaire O, Richards JG, Faull RL, Schweizer A (1999) XCE, a new member of the endothelin-converting enzyme and neutral endopeptidase family, is preferentially expressed in the CNS. Brain Res Mol Brain Res 64:211–221CrossRefPubMedGoogle Scholar
  32. 32.
    Wichterle H, Lieberam I, Porter JA, Jessell TM (2002) Directed differentiation of embryonic stem cells into motor neurons. Cell 110:385–397CrossRefPubMedGoogle Scholar
  33. 33.
    Williams MS, Elliott CG, Bamshad MJ (2007) Pulmonary disease is a component of distal arthrogryposis type 5. Am J Med Genet A 143A:752–756CrossRefPubMedGoogle Scholar
  34. 34.
    Yang H, Wang H, Jaenisch R (2014) Generating genetically modified mice using CRISPR/Cas-mediated genome engineering. Nat Protoc 9:1956–1968. doi: 10.1038/nprot.2014.134 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Laboratory for Proteolytic NeuroscienceRIKEN Brain Science InstituteSaitamaJapan
  2. 2.Department of Functional Anatomy and Neuroscience, Graduate School of MedicineNagoya UniversityNagoyaJapan

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