Regulation of neurofilament dynamics by phosphorylation

Neurofilament (NF) phosphorylation has long been considered to regulate axonal transport rate and in doing so to provide stability to mature axons. Studies utilizing mice in which the C‐terminal region of NF subunits (which contains the vast majority of phosphorylation sites) has been deleted has prompted an ongoing challenge to this hypothesis. We evaluate the collective evidence to date for and against a role for NF C‐terminal phosphorylation in regulation of axonal transport and in providing structural support for axons, including some novel studies from our laboratory. We present a few suggestions for further experimentation in this area, and expand upon previous models for axonal NF dynamics. Finally, we address how C‐terminal phosphorylation is regionally and temporally regulated by a balance of kinase and phosphatase activities, and how misregulation of this balance can contribute to motor neuron disease.

[1]  N. Calcutt,et al.  Gene replacement in mice reveals that the heavily phosphorylated tail of neurofilament heavy subunit does not affect axonal caliber or the transit of cargoes in slow axonal transport , 2002, The Journal of cell biology.

[2]  T B Shea,et al.  Phospho-dependent association of neurofilament proteins with kinesin in situ. , 2000, Cell motility and the cytoskeleton.

[3]  Veeranna,et al.  Phosphorylation of the Head Domain of Neurofilament Protein (NF-M) , 2003, Journal of Biological Chemistry.

[4]  T. Shea,et al.  C-terminal phosphorylation of the high molecular weight neurofilament subunit correlates with decreased neurofilament axonal transport velocity , 2000, Brain Research.

[5]  Lei Wang,et al.  Stochastic simulation of neurofilament transport in axons: the "stop-and-go" hypothesis. , 2005, Molecular biology of the cell.

[6]  P. Janmey,et al.  Bidirectional translocation of neurofilaments along microtubules mediated in part by dynein/dynactin. , 2000, Molecular biology of the cell.

[7]  C. Shaw,et al.  Neurofilament heavy chain side arm phosphorylation regulates axonal transport of neurofilaments , 2003, The Journal of cell biology.

[8]  C. Theiss,et al.  Impairment of anterograde and retrograde neurofilament transport after anti-kinesin and anti-dynein antibody microinjection in chicken dorsal root ganglia. , 2005, European journal of cell biology.

[9]  M. Strong,et al.  Phosphorylation state of the native high‐molecular‐weight neurofilament subunit protein from cervical spinal cord in sporadic amyotrophic lateral sclerosis , 2001, Journal of neurochemistry.

[10]  M. Black,et al.  Role of cytoplasmic dynein in the axonal transport of microtubules and neurofilaments , 2005, The Journal of cell biology.

[11]  R. Nixon,et al.  The slow axonal transport of cytoskeletal proteins. , 1998, Current opinion in cell biology.

[12]  Xinran Liu,et al.  Abnormal neurofilament transport caused by targeted disruption of neuronal kinesin heavy chain KIF5A , 2003, The Journal of cell biology.

[13]  H. Pant,et al.  Mitogen-activated protein kinase regulates neurofilament axonal transport , 2004, Journal of Cell Science.

[14]  Kazuko Hasegawa,et al.  Kinesin and cytoplasmic dynein in spinal spheroids with motor neuron disease , 1998, Journal of the Neurological Sciences.

[15]  R. Goldman,et al.  Cdk5 regulates axonal transport and phosphorylation of neurofilaments in cultured neurons , 2004, Journal of Cell Science.

[16]  E. Wheeler,et al.  Aluminum inhibits neurofilament assembly, cytoskeletal incorporation, and axonal transport. Dynamic nature of aluminum-induced perikaryal neurofilament accumulations as revealed by subunit turnover. , 1997, Molecular and chemical neuropathology.

[17]  Jean-Pierre Julien,et al.  Neurofilaments in health and disease. , 1998, Progress in nucleic acid research and molecular biology.

[18]  William J. Beaty,et al.  Traffic Jams: Dynamic Models for Neurofilament Accumulation in Motor Neuron Disease , 2007, Traffic.

[19]  R. Nixon,et al.  The Regulation of Neurofilament Protein Dynamics by Phosphorylation: Clues to Neurofibrillary Pathobiology , 1993, Brain pathology.

[20]  D. Price,et al.  Slowing of the axonal transport of neurofilament proteins during development , 1983, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[21]  T. Shea,et al.  Dynein mediates retrograde neurofilament transport within axons and anterograde delivery of NFs from perikarya into axons: regulation by multiple phosphorylation events. , 2006, Cell motility and the cytoskeleton.

[22]  P. Leigh,et al.  Cyclin dependent kinase-5 (CDK-5) phosphorylates neurofilament heavy (NF-H) chain to generate epitopes for antibodies that label neurofilament accumulations in amyotrophic lateral sclerosis (ALS) and is present in affected motor neurones in ALS , 1999, Progress in Neuro-Psychopharmacology and Biological Psychiatry.

[23]  Ram K. Sihag,et al.  Local Control of Neurofilament Accumulation during Radial Growth of Myelinating Axons in Vivo , 2000, The Journal of cell biology.

[24]  D. Cleveland,et al.  Altered axonal architecture by removal of the heavily phosphorylated neurofilament tail domains strongly slows superoxide dismutase 1 mutant-mediated ALS. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[25]  T. Shea,et al.  Occam's Razor Slices Through the Mysteries of Neurofilament Axonal Transport: Can it Really be so Simple? , 2000, Traffic.

[26]  J. Trojanowski,et al.  Overexpression of the human NFM subunit in transgenic mice modifies the level of endogenous NFL and the phosphorylation state of NFH subunits , 1995, The Journal of cell biology.

[27]  M. Black,et al.  Phosphorylation of neurofilament proteins in intact neurons: demonstration of phosphorylation in cell bodies and axons , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[28]  J J Blum,et al.  A model for slow axonal transport and its application to neurofilamentous neuropathies. , 1989, Cell motility and the cytoskeleton.

[29]  J. Julien,et al.  Disruption of the NF-H Gene Increases Axonal Microtubule Content and Velocity of Neurofilament Transport: Relief of Axonopathy Resulting from the Toxin β,β′-Iminodipropionitrile , 1998, The Journal of cell biology.

[30]  R. Lasek,et al.  Slow axonal transport mechanisms move neurofilaments relentlessly in mouse optic axons , 1992, The Journal of cell biology.

[31]  T. Shea,et al.  The protein phosphatase inhibitor okadaic acid increases axonal neurofilaments and neurite caliber, and decreases axonal microtubules in NB2a/d1 Cells , 1993, Journal of neuroscience research.

[32]  Torsten Wittmann,et al.  Motor proteins regulate force interactions between microtubules and microfilaments in the axon , 2000, Nature Cell Biology.

[33]  Lisa A Flanagan,et al.  Kinesin, dynein and neurofilament transport , 2001, Trends in Neurosciences.

[34]  T B Shea,et al.  Kinesin-mediated transport of neurofilament protein oligomers in growing axons. , 1999, Journal of cell science.

[35]  Avner Friedman,et al.  A dynamical system model of neurofilament transport in axons. , 2005, Journal of theoretical biology.

[36]  R. Nixon,et al.  Deleting the phosphorylated tail domain of the neurofilament heavy subunit does not alter neurofilament transport rate in vivo , 2006, Neuroscience Letters.

[37]  T. Shea,et al.  Neurofilament subunits undergo more rapid translocation within retinas than in optic axons. , 2004, Brain research. Molecular brain research.

[38]  R. Lasek,et al.  The maximum rate of neurofilament transport in axons: a view of molecular transport mechanisms continuously engaged , 1993, Brain Research.

[39]  T. Shea,et al.  Neurofilaments Consist of Distinct Populations That Can Be Distinguished by C-Terminal Phosphorylation, Bundling, and Axonal Transport Rate in Growing Axonal Neurites , 2001, The Journal of Neuroscience.

[40]  W. Saxton,et al.  Cytoplasmic dynein, the dynactin complex, and kinesin are interdependent and essential for fast axonal transport. , 1999, Molecular biology of the cell.

[41]  P. Baas,et al.  Axonal Transport of Microtubules: the Long and Short of It , 2006, Traffic.

[42]  T. Gotow,et al.  The neurofilament middle molecular mass subunit carboxyl-terminal tail domains is essential for the radial growth and cytoskeletal architecture of axons but not for regulating neurofilament transport rate , 2003, The Journal of cell biology.

[43]  M. Sacher,et al.  Phosphorylation of Neurofilament Proteins , 1995 .

[44]  R. Nixon,et al.  Defective Neurofilament Transport in Mouse Models of Amyotrophic Lateral Sclerosis: A Review , 2003, Neurochemical Research.

[45]  C. Shaw,et al.  p38alpha stress-activated protein kinase phosphorylates neurofilaments and is associated with neurofilament pathology in amyotrophic lateral sclerosis. , 2004, Molecular and cellular neurosciences.

[46]  T. Shea,et al.  Regulation of neurofilament axonal transport by phosphorylation in optic axons in situ. , 1999, Cell motility and the cytoskeleton.

[47]  Veeranna,et al.  α-Internexin Is Structurally and Functionally Associated with the Neurofilament Triplet Proteins in the Mature CNS , 2006, The Journal of Neuroscience.

[48]  J. Julien,et al.  Defective axonal transport in a transgenic mouse model of amyotrophic lateral sclerosis , 1995, Nature.

[49]  Lei Wang,et al.  Rapid movement of axonal neurofilaments interrupted by prolonged pauses , 2000, Nature Cell Biology.

[50]  J. Trojanowski,et al.  Neurofilaments and Orthograde Transport Are Reduced in Ventral Root Axons of Transgenic Mice that Express Human SOD1 with a G93A Mutation , 1997, The Journal of cell biology.

[51]  P. Janmey,et al.  Regulation of neurofilament interactionsin vitro by natural and synthetic polypeptides sharing Lys-Ser-Pro sequences with the heavy neurofilament subunit NF-H: Neurofilament crossbridging by antiparallel sidearm overlapping , 1998, Medical and Biological Engineering and Computing.

[52]  Jonathan D Cooper,et al.  p38α stress-activated protein kinase phosphorylates neurofilaments and is associated with neurofilament pathology in amyotrophic lateral sclerosis , 2004, Molecular and Cellular Neuroscience.

[53]  R. Nixon,et al.  Multiple fates of newly synthesized neurofilament proteins: evidence for a stationary neurofilament network distributed nonuniformly along axons of retinal ganglion cell neurons , 1986, The Journal of cell biology.

[54]  H. Pant,et al.  Does neurofilament phosphorylation regulate axonal transport? , 2003, Trends in Neurosciences.

[55]  Anthony Brown,et al.  Contiguous phosphorylated and non-phosphorylated domains along axonal neurofilaments. , 1998, Journal of cell science.

[56]  R. H. Brown,et al.  A transgenic-mouse model of amyotrophic lateral sclerosis. , 1994, The New England journal of medicine.

[57]  Scott T. Brady,et al.  Local modulation of neurofilament phosphorylation, axonal caliber, and slow axonal transport by myelinating Schwann cells , 1992, Cell.

[58]  M. Inagaki,et al.  Role of phosphorylation on the structural dynamics and function of types III and IV intermediate filaments. , 2007, Experimental cell research.

[59]  Scott T. Brady,et al.  Neurofilaments Are Transported Rapidly But Intermittently in Axons: Implications for Slow Axonal Transport , 2000, The Journal of Neuroscience.

[60]  T. Crawford,et al.  Neurofilament subunit NF-H modulates axonal diameter by selectively slowing neurofilament transport , 1996, The Journal of cell biology.

[61]  T. Shea,et al.  Neurofilaments can undergo axonal transport and cytoskeletal incorporation in a discontinuous manner. , 2005, Cell motility and the cytoskeleton.

[62]  J. Griffin,et al.  Phosphorylation‐Dependent Immunoreactivity of Neurofilaments and the Rate of Slow Axonal Transport in the Central and Peripheral Axons of the Rat Dorsal Root Ganglion , 1994, Journal of neurochemistry.

[63]  J. Glass,et al.  Redistribution of cytoskeletal proteins in mammalian axons disconnected from their cell bodies , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[64]  Sangmook Lee,et al.  Hypophosphorylated neurofilament subunits undergo axonal transport more rapidly than more extensively phosphorylated subunits in situ. , 2000, Cell motility and the cytoskeleton.

[65]  R. Nixon,et al.  Multiple phosphorylated variants of the high molecular mass subunit of neurofilaments in axons of retinal cell neurons: characterization and evidence for their differential association with stationary and moving neurofilaments , 1988, The Journal of cell biology.

[66]  Veeranna,et al.  Neurofilament phosphorylation. , 1995, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[67]  T. Shea,et al.  Selective accumulation of the high molecular weight neurofilament subunit within the distal region of growing axonal neurites. , 2001, Cell motility and the cytoskeleton.

[68]  B. Helfand,et al.  Fast transport of neurofilament protein along microtubules in squid axoplasm. , 2000, Journal of cell science.

[69]  Anthony Brown,et al.  Slow Axonal Transport of Neurofilament Protein in Cultured Neurons , 1999, The Journal of cell biology.

[70]  Sangmook Lee,et al.  The high and middle molecular weight neurofilament subunits regulate the association of neurofilaments with kinesin: inhibition by phosphorylation of the high molecular weight subunit. , 2005, Brain research. Molecular brain research.

[71]  Peter Jung,et al.  Neurofilaments Switch between Distinct Mobile and Stationary States during Their Transport along Axons , 2007, The Journal of Neuroscience.

[72]  Po-Wen Chen,et al.  Divergent effects of the MEKK-1/JNK pathway on NB2a/d1 differentiation: Some activity is required for outgrowth and stabilization of neurites but overactivation inhibits both phenomena , 2006, Brain Research.

[73]  M. Sugawara,et al.  Kinesin accumulation in chick spinal axonal swellings with β,β′-iminodipropionitrile (IDPN) intoxication , 1998, Neuroscience Letters.