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. 1996 Nov 1;135(3):711–724. doi: 10.1083/jcb.135.3.711

Neurofilament subunit NF-H modulates axonal diameter by selectively slowing neurofilament transport

PMCID: PMC2121055  PMID: 8909545

Abstract

To examine the mechanism through which neurofilaments regulate the caliber of myelinated axons and to test how aberrant accumulations of neurofilaments cause motor neuron disease, mice have been constructed that express wild-type mouse NF-H up to 4.5 times the normal level. Small increases in NF-H expression lead to increased total neurofilament content and larger myelinated axons, whereas larger increases in NF-H decrease total neurofilament content and strongly inhibit radial growth. Increasing NF-H expression selectively slow neurofilament transport into and along axons, resulting in severe perikaryal accumulation of neurofilaments and proximal axonal swellings in motor neurons. Unlike the situation in transgenic mice expressing modest levels of human NF-H (Cote, F., J.F. Collard, and J.P. Julien. 1993. Cell. 73:35-46), even 4.5 times the normal level of wild-type mouse NF-H does not result in any overt phenotype or enhanced motor neuron degeneration or loss. Rather, motor neurons are extraordinarily tolerant of wild-type murine NF-H, whereas wild-type human NF-H, which differs from the mouse homolog at > 160 residue positions, mediates motor neuron disease in mice by acting as an aberrant, mutant subunit.

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Selected References

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  1. Arbuthnott E. R., Boyd I. A., Kalu K. U. Ultrastructural dimensions of myelinated peripheral nerve fibres in the cat and their relation to conduction velocity. J Physiol. 1980 Nov;308:125–157. doi: 10.1113/jphysiol.1980.sp013465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckman J. S., Carson M., Smith C. D., Koppenol W. H. ALS, SOD and peroxynitrite. Nature. 1993 Aug 12;364(6438):584–584. doi: 10.1038/364584a0. [DOI] [PubMed] [Google Scholar]
  3. Brown A., Bernier G., Mathieu M., Rossant J., Kothary R. The mouse dystonia musculorum gene is a neural isoform of bullous pemphigoid antigen 1. Nat Genet. 1995 Jul;10(3):301–306. doi: 10.1038/ng0795-301. [DOI] [PubMed] [Google Scholar]
  4. Carpenter S. Proximal axonal enlargement in motor neuron disease. Neurology. 1968 Sep;18(9):841–851. doi: 10.1212/wnl.18.9.841. [DOI] [PubMed] [Google Scholar]
  5. Ching G. Y., Liem R. K. Assembly of type IV neuronal intermediate filaments in nonneuronal cells in the absence of preexisting cytoplasmic intermediate filaments. J Cell Biol. 1993 Sep;122(6):1323–1335. doi: 10.1083/jcb.122.6.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark A. W., Griffin J. W., Price D. L. The axonal pathology in chronic IDPN intoxication. J Neuropathol Exp Neurol. 1980 Jan;39(1):42–55. doi: 10.1097/00005072-198001000-00004. [DOI] [PubMed] [Google Scholar]
  7. Cleveland D. W., Monteiro M. J., Wong P. C., Gill S. R., Gearhart J. D., Hoffman P. N. Involvement of neurofilaments in the radial growth of axons. J Cell Sci Suppl. 1991;15:85–95. doi: 10.1242/jcs.1991.supplement_15.12. [DOI] [PubMed] [Google Scholar]
  8. Collard J. F., Côté F., Julien J. P. Defective axonal transport in a transgenic mouse model of amyotrophic lateral sclerosis. Nature. 1995 May 4;375(6526):61–64. doi: 10.1038/375061a0. [DOI] [PubMed] [Google Scholar]
  9. Côté F., Collard J. F., Julien J. P. Progressive neuronopathy in transgenic mice expressing the human neurofilament heavy gene: a mouse model of amyotrophic lateral sclerosis. Cell. 1993 Apr 9;73(1):35–46. doi: 10.1016/0092-8674(93)90158-m. [DOI] [PubMed] [Google Scholar]
  10. Eyer J., Peterson A. Neurofilament-deficient axons and perikaryal aggregates in viable transgenic mice expressing a neurofilament-beta-galactosidase fusion protein. Neuron. 1994 Feb;12(2):389–405. doi: 10.1016/0896-6273(94)90280-1. [DOI] [PubMed] [Google Scholar]
  11. Figlewicz D. A., Krizus A., Martinoli M. G., Meininger V., Dib M., Rouleau G. A., Julien J. P. Variants of the heavy neurofilament subunit are associated with the development of amyotrophic lateral sclerosis. Hum Mol Genet. 1994 Oct;3(10):1757–1761. doi: 10.1093/hmg/3.10.1757. [DOI] [PubMed] [Google Scholar]
  12. Friede R. L., Samorajski T. Axon caliber related to neurofilaments and microtubules in sciatic nerve fibers of rats and mice. Anat Rec. 1970 Aug;167(4):379–387. doi: 10.1002/ar.1091670402. [DOI] [PubMed] [Google Scholar]
  13. Guo L., Degenstein L., Dowling J., Yu Q. C., Wollmann R., Perman B., Fuchs E. Gene targeting of BPAG1: abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degeneration. Cell. 1995 Apr 21;81(2):233–243. doi: 10.1016/0092-8674(95)90333-x. [DOI] [PubMed] [Google Scholar]
  14. Hirano A. Cytopathology of amyotrophic lateral sclerosis. Adv Neurol. 1991;56:91–101. [PubMed] [Google Scholar]
  15. Hirano A., Donnenfeld H., Sasaki S., Nakano I. Fine structural observations of neurofilamentous changes in amyotrophic lateral sclerosis. J Neuropathol Exp Neurol. 1984 Sep;43(5):461–470. doi: 10.1097/00005072-198409000-00001. [DOI] [PubMed] [Google Scholar]
  16. Hirano A., Nakano I., Kurland L. T., Mulder D. W., Holley P. W., Saccomanno G. Fine structural study of neurofibrillary changes in a family with amyotrophic lateral sclerosis. J Neuropathol Exp Neurol. 1984 Sep;43(5):471–480. doi: 10.1097/00005072-198409000-00002. [DOI] [PubMed] [Google Scholar]
  17. Hirokawa N., Glicksman M. A., Willard M. B. Organization of mammalian neurofilament polypeptides within the neuronal cytoskeleton. J Cell Biol. 1984 Apr;98(4):1523–1536. doi: 10.1083/jcb.98.4.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoffman P. N., Cleveland D. W., Griffin J. W., Landes P. W., Cowan N. J., Price D. L. Neurofilament gene expression: a major determinant of axonal caliber. Proc Natl Acad Sci U S A. 1987 May;84(10):3472–3476. doi: 10.1073/pnas.84.10.3472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hoffman P. N., Griffin J. W., Price D. L. Control of axonal caliber by neurofilament transport. J Cell Biol. 1984 Aug;99(2):705–714. doi: 10.1083/jcb.99.2.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoffman P. N., Lasek R. J., Griffin J. W., Price D. L. Slowing of the axonal transport of neurofilament proteins during development. J Neurosci. 1983 Aug;3(8):1694–1700. doi: 10.1523/JNEUROSCI.03-08-01694.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hoffman P. N., Lasek R. J. The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons. J Cell Biol. 1975 Aug;66(2):351–366. doi: 10.1083/jcb.66.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hsieh S. T., Kidd G. J., Crawford T. O., Xu Z., Lin W. M., Trapp B. D., Cleveland D. W., Griffin J. W. Regional modulation of neurofilament organization by myelination in normal axons. J Neurosci. 1994 Nov;14(11 Pt 1):6392–6401. doi: 10.1523/JNEUROSCI.14-11-06392.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Julien J. P., Côté F., Beaudet L., Sidky M., Flavell D., Grosveld F., Mushynski W. Sequence and structure of the mouse gene coding for the largest neurofilament subunit. Gene. 1988 Sep 7;68(2):307–314. doi: 10.1016/0378-1119(88)90033-9. [DOI] [PubMed] [Google Scholar]
  24. Lasek R. J., Paggi P., Katz M. J. Slow axonal transport mechanisms move neurofilaments relentlessly in mouse optic axons. J Cell Biol. 1992 May;117(3):607–616. doi: 10.1083/jcb.117.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lee M. K., Cleveland D. W. Neurofilament function and dysfunction: involvement in axonal growth and neuronal disease. Curr Opin Cell Biol. 1994 Feb;6(1):34–40. doi: 10.1016/0955-0674(94)90113-9. [DOI] [PubMed] [Google Scholar]
  26. Lee M. K., Marszalek J. R., Cleveland D. W. A mutant neurofilament subunit causes massive, selective motor neuron death: implications for the pathogenesis of human motor neuron disease. Neuron. 1994 Oct;13(4):975–988. doi: 10.1016/0896-6273(94)90263-1. [DOI] [PubMed] [Google Scholar]
  27. Lee M. K., Tuttle J. B., Rebhun L. I., Cleveland D. W., Frankfurter A. The expression and posttranslational modification of a neuron-specific beta-tubulin isotype during chick embryogenesis. Cell Motil Cytoskeleton. 1990;17(2):118–132. doi: 10.1002/cm.970170207. [DOI] [PubMed] [Google Scholar]
  28. Lee M. K., Xu Z., Wong P. C., Cleveland D. W. Neurofilaments are obligate heteropolymers in vivo. J Cell Biol. 1993 Sep;122(6):1337–1350. doi: 10.1083/jcb.122.6.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lee V. M., Carden M. J., Schlaepfer W. W., Trojanowski J. Q. Monoclonal antibodies distinguish several differentially phosphorylated states of the two largest rat neurofilament subunits (NF-H and NF-M) and demonstrate their existence in the normal nervous system of adult rats. J Neurosci. 1987 Nov;7(11):3474–3488. doi: 10.1523/JNEUROSCI.07-11-03474.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lees J. F., Shneidman P. S., Skuntz S. F., Carden M. J., Lazzarini R. A. The structure and organization of the human heavy neurofilament subunit (NF-H) and the gene encoding it. EMBO J. 1988 Jul;7(7):1947–1955. doi: 10.1002/j.1460-2075.1988.tb03032.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lopata M. A., Cleveland D. W. In vivo microtubules are copolymers of available beta-tubulin isotypes: localization of each of six vertebrate beta-tubulin isotypes using polyclonal antibodies elicited by synthetic peptide antigens. J Cell Biol. 1987 Oct;105(4):1707–1720. doi: 10.1083/jcb.105.4.1707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Monteiro M. J., Hoffman P. N., Gearhart J. D., Cleveland D. W. Expression of NF-L in both neuronal and nonneuronal cells of transgenic mice: increased neurofilament density in axons without affecting caliber. J Cell Biol. 1990 Oct;111(4):1543–1557. doi: 10.1083/jcb.111.4.1543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nixon R. A., Logvinenko K. B. Multiple fates of newly synthesized neurofilament proteins: evidence for a stationary neurofilament network distributed nonuniformly along axons of retinal ganglion cell neurons. J Cell Biol. 1986 Feb;102(2):647–659. doi: 10.1083/jcb.102.2.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nixon R. A., Paskevich P. A., Sihag R. K., Thayer C. Y. Phosphorylation on carboxyl terminus domains of neurofilament proteins in retinal ganglion cell neurons in vivo: influences on regional neurofilament accumulation, interneurofilament spacing, and axon caliber. J Cell Biol. 1994 Aug;126(4):1031–1046. doi: 10.1083/jcb.126.4.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ohara O., Gahara Y., Miyake T., Teraoka H., Kitamura T. Neurofilament deficiency in quail caused by nonsense mutation in neurofilament-L gene. J Cell Biol. 1993 Apr;121(2):387–395. doi: 10.1083/jcb.121.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rooke K., Figlewicz D. A., Han F. Y., Rouleau G. A. Analysis of the KSP repeat of the neurofilament heavy subunit in familiar amyotrophic lateral sclerosis. Neurology. 1996 Mar;46(3):789–790. doi: 10.1212/wnl.46.3.789. [DOI] [PubMed] [Google Scholar]
  37. Rosen D. R., Siddique T., Patterson D., Figlewicz D. A., Sapp P., Hentati A., Donaldson D., Goto J., O'Regan J. P., Deng H. X. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature. 1993 Mar 4;362(6415):59–62. doi: 10.1038/362059a0. [DOI] [PubMed] [Google Scholar]
  38. Rouleau G. A., Clark A. W., Rooke K., Pramatarova A., Krizus A., Suchowersky O., Julien J. P., Figlewicz D. SOD1 mutation is associated with accumulation of neurofilaments in amyotrophic lateral sclerosis. Ann Neurol. 1996 Jan;39(1):128–131. doi: 10.1002/ana.410390119. [DOI] [PubMed] [Google Scholar]
  39. Sakaguchi T., Okada M., Kitamura T., Kawasaki K. Reduced diameter and conduction velocity of myelinated fibers in the sciatic nerve of a neurofilament-deficient mutant quail. Neurosci Lett. 1993 Apr 16;153(1):65–68. doi: 10.1016/0304-3940(93)90078-y. [DOI] [PubMed] [Google Scholar]
  40. Schlaepfer W. W., Bruce J. Neurofilament proteins are distributed in a diminishing proximodistal gradient along rat sciatic nerve. J Neurochem. 1990 Aug;55(2):453–460. doi: 10.1111/j.1471-4159.1990.tb04157.x. [DOI] [PubMed] [Google Scholar]
  41. Shibata N., Hirano A., Kobayashi M., Siddique T., Deng H. X., Hung W. Y., Kato T., Asayama K. Intense superoxide dismutase-1 immunoreactivity in intracytoplasmic hyaline inclusions of familial amyotrophic lateral sclerosis with posterior column involvement. J Neuropathol Exp Neurol. 1996 Apr;55(4):481–490. doi: 10.1097/00005072-199604000-00011. [DOI] [PubMed] [Google Scholar]
  42. Troncoso J. C., March J. L., Häner M., Aebi U. Effect of aluminum and other multivalent cations on neurofilaments in vitro: an electron microscopic study. J Struct Biol. 1990 Mar;103(1):2–12. doi: 10.1016/1047-8477(90)90080-v. [DOI] [PubMed] [Google Scholar]
  43. Wiedau-Pazos M., Goto J. J., Rabizadeh S., Gralla E. B., Roe J. A., Lee M. K., Valentine J. S., Bredesen D. E. Altered reactivity of superoxide dismutase in familial amyotrophic lateral sclerosis. Science. 1996 Jan 26;271(5248):515–518. doi: 10.1126/science.271.5248.515. [DOI] [PubMed] [Google Scholar]
  44. Willard M., Simon C. Modulations of neurofilament axonal transport during the development of rabbit retinal ganglion cells. Cell. 1983 Dec;35(2 Pt 1):551–559. doi: 10.1016/0092-8674(83)90189-7. [DOI] [PubMed] [Google Scholar]
  45. Wong P. C., Marszalek J., Crawford T. O., Xu Z., Hsieh S. T., Griffin J. W., Cleveland D. W. Increasing neurofilament subunit NF-M expression reduces axonal NF-H, inhibits radial growth, and results in neurofilamentous accumulation in motor neurons. J Cell Biol. 1995 Sep;130(6):1413–1422. doi: 10.1083/jcb.130.6.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wong P. C., Pardo C. A., Borchelt D. R., Lee M. K., Copeland N. G., Jenkins N. A., Sisodia S. S., Cleveland D. W., Price D. L. An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria. Neuron. 1995 Jun;14(6):1105–1116. doi: 10.1016/0896-6273(95)90259-7. [DOI] [PubMed] [Google Scholar]
  47. Xu Z., Cork L. C., Griffin J. W., Cleveland D. W. Increased expression of neurofilament subunit NF-L produces morphological alterations that resemble the pathology of human motor neuron disease. Cell. 1993 Apr 9;73(1):23–33. doi: 10.1016/0092-8674(93)90157-l. [DOI] [PubMed] [Google Scholar]
  48. Xu Z., Marszalek J. R., Lee M. K., Wong P. C., Folmer J., Crawford T. O., Hsieh S. T., Griffin J. W., Cleveland D. W. Subunit composition of neurofilaments specifies axonal diameter. J Cell Biol. 1996 Jun;133(5):1061–1069. doi: 10.1083/jcb.133.5.1061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Yamasaki H., Bennett G. S., Itakura C., Mizutani M. Defective expression of neurofilament protein subunits in hereditary hypotrophic axonopathy of quail. Lab Invest. 1992 Jun;66(6):734–743. [PubMed] [Google Scholar]
  50. Yang Y., Dowling J., Yu Q. C., Kouklis P., Cleveland D. W., Fuchs E. An essential cytoskeletal linker protein connecting actin microfilaments to intermediate filaments. Cell. 1996 Aug 23;86(4):655–665. doi: 10.1016/s0092-8674(00)80138-5. [DOI] [PubMed] [Google Scholar]
  51. de Waegh S. M., Lee V. M., Brady S. T. Local modulation of neurofilament phosphorylation, axonal caliber, and slow axonal transport by myelinating Schwann cells. Cell. 1992 Feb 7;68(3):451–463. doi: 10.1016/0092-8674(92)90183-d. [DOI] [PubMed] [Google Scholar]

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