Acta Neuropathologica

, Volume 119, Issue 2, pp 235–248

Paclitaxel induces axonal microtubules polar reconfiguration and impaired organelle transport: implications for the pathogenesis of paclitaxel-induced polyneuropathy

Original Paper


In differentiated axons almost all microtubules (MTs) uniformly point their plus ends towards the axonal tip. The uniform polar pattern provides the structural substrate for efficient organelle transport along axons. It is generally believed that the mass and pattern of MTs polar orientation remain unchanged in differentiated neurons. Here we examined long-term effects of the MTs stabilizing reagent paclitaxel (taxol) over MTs polar orientation and organelle transport in cultured Aplysia neurons. Unexpectedly, we found that rather than stabilizing the MTs, paclitaxel leads to their massive polar reconfiguration, accompanied by impaired organelle transport. Washout of paclitaxel does not lead to recovery of the polar orientation indicating that the new pattern is self-maintained. Taken together the data suggest that MTs in differentiated neurons maintain the potential to be reconfigured. Such reconfiguration may serve physiological functions or lead to degeneration. In addition, our observations offer a novel mechanism that could account for the development of peripheral neuropathy in patients receiving paclitaxel as an antitumor drug.


Paclitaxel Microtubules Plus ends binding proteins Axoplasmic transport Peripheral neuropathy Aplysia 

Supplementary material

401_2009_586_MOESM1_ESM.doc (35 kb)
Supplementary material 1 (DOC 35 kb)
401_2009_586_MOESM2_ESM.doc (36 kb)
Supplementary material 2 (DOC 37 kb)

Movie S1. Paclitaxel induced MT polar reconfiguration. (A) EB3-GFP comet tails in control axon (before paclitaxel application), 100-200µm away from cell body. (B) The same axon, at the same location 72h after the application of 100nM paclitaxel into the bathing solution. The video contains 30 images, taken at intervals of 5.7 seconds. The frames are shown at a rate of 10/s. Scalebar: 10 µm (MPG 12.6 MB)

401_2009_586_MOESM4_ESM.mpg (8.5 mb)
Movie S2. Incubation of a neuron with 10nM paclitaxel. (A) EB3-GFP comet tails in a control axon (before application of paclitaxel). (B) The same axon, at the same location 96h after the addition of 10nM paclitaxel to the bathing solution. The video contains 30 images, taken at intervals of 5.7 seconds. The frames are shown at a rate of 10/s. Scalebar: 10 µm (MPG 8.52 MB)
401_2009_586_MOESM5_ESM.mpg (9.8 mb)
Movie S3. EB3-GFP comet tails and retrograde transport in a control neuron. (A) EB3-GFP comet tails in a control neuron (green), 200-300µm from cell body and (B) transport of SR101 labeled organells (red). The videos comprise 30 images, taken at intervals of 4.5 seconds. Scalebar: 10 µm (MPG 9.84 MB)
401_2009_586_MOESM6_ESM.mpg (9.8 mb)
Movie S4. 10nM paclitaxel leads to impaired transport of SR101 labeled organells. (A) EB3-GFP (green) comet tails in a neuron 72h after the onset of 100nM paclitaxel incubation, 200-300µm from cell body. (B) The transport of SR101 labeled pinocytotic organelles (red). The video comprises 30 images, taken at intervals of 5.7 seconds. Scalebar: 10 µm (MPG 9.84 MB)

Movie S5. 100nM paclitaxel leads to severe impairments in SR101 labeled vesicles. (A) The video (green) depicts EB3-GFP comet tails in a neuron 48h after the onset of 100nM paclitaxel incubation, 200-300µm from cell body. And (B), the transport of SR101 labeled pinocytotic vesicles. The video comprises 30 images, taken at intervals of 5.5 seconds. Scalebar: 10 µm (MPG 12.6 MB)

Movie S6. 100nM paclitaxel impedes mitochondrial transport. (A) A buccal neuron was injected with hOACTL-GFP mRNA and imaged 24h later. 25 frames of hOACTL-GFP labeled mitochondria were taken 5.3s apart. (B) Thereafter, the neuron was incubated with 100nM paclitaxel and imaged 24h later. 25 frames were taken at an interval of 5.3s. (C) The axon 72h following paclitaxel incubation. 25 frames were taken at an interval of 5.3s. Scalebar: 10µm (MPG 7.64 MB)

Movie S7. Washout of paclitaxel does not lead to recovery of the MTs polar pattern. The video depicts the same neuron presented in Fig. 8, 48h following 100nM paclitaxel washout. (A) EB3-GFP (green) comet tails in a neuron, 200-300µm from cell body. (B) The transport of SR101 labeled organells. The video comprises 30 images, taken at intervals of 5.5 seconds. Scalebar: 10 µm (MPG 12.6 MB)


  1. 1.
    Argyriou AA, Koltzenburg M, Polychronopoulos P, Papapetropoulos S, Kalofonos HP (2008) Peripheral nerve damage associated with administration of taxanes in patients with cancer. Crit Rev Oncol Hematol 66:218–228CrossRefPubMedGoogle Scholar
  2. 2.
    Baas PW, Black MM, Banker GA (1989) Changes in microtubule polarity orientation during the development of hippocampal neurons in culture. J Cell Biol 109:3085–3094CrossRefPubMedGoogle Scholar
  3. 3.
    Baas PW, Karabay A, Qiang L (2005) Microtubules cut and run. Trends Cell Biol 15:518–524CrossRefPubMedGoogle Scholar
  4. 4.
    Burton PR (1988) Dendrites of mitral cell neurons contain microtubules of opposite polarity. Brain Res 473:107–115CrossRefPubMedGoogle Scholar
  5. 5.
    Burton PR, Paige JL (1981) Polarity of axoplasmic microtubules in the olfactory nerve of the frog. Proc Natl Acad Sci USA 78:3269–3273CrossRefPubMedGoogle Scholar
  6. 6.
    Callizot N, Andriambeloson E, Glass J et al (2008) Interleukin-6 protects against paclitaxel, cisplatin and vincristine-induced neuropathies without impairing chemotherapeutic activity. Cancer Chemother Pharmacol 62:995–1007CrossRefPubMedGoogle Scholar
  7. 7.
    Conde C, Caceres A (2009) Microtubule assembly, organization and dynamics in axons and dendrites. Nat Rev Neurosci 10:319–332CrossRefPubMedGoogle Scholar
  8. 8.
    Cytrynbaum EN, Rodionov V, Mogilner A (2004) Computational model of dynein-dependent self-organization of microtubule asters. J Cell Sci 117:1381–1397CrossRefPubMedGoogle Scholar
  9. 9.
    Cytrynbaum EN, Rodionov V, Mogilner A (2006) Nonlocal mechanism of self-organization and centering of microtubule asters. Bull Math Biol 68:1053–1072CrossRefPubMedGoogle Scholar
  10. 10.
    De Brabander M, Geuens G, Nuydens R, Willebrords R, De Mey J (1981) Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci USA 78:5608–5612CrossRefPubMedGoogle Scholar
  11. 11.
    De Vos KJ, Grierson AJ, Ackerley S, Miller CC (2008) Role of axonal transport in neurodegenerative diseases. Annu Rev Neurosci 31:151–173CrossRefPubMedGoogle Scholar
  12. 12.
    Dehmelt L, Nalbant P, Steffen W, Halpain S (2006) A microtubule-based, dynein-dependent force induces local cell protrusions: implications for neurite initiation. Brain Cell Biol 35:39–56CrossRefPubMedGoogle Scholar
  13. 13.
    Erez H, Malkinson G, Prager-Khoutorsky M et al (2007) Formation of microtubule-based traps controls the sorting and concentration of vesicles to restricted sites of regenerating neurons after axotomy. J Cell Biol 176:497–507CrossRefPubMedGoogle Scholar
  14. 14.
    Erez H, Spira ME (2008) Local self-assembly mechanisms underlie the differential transformation of the proximal and distal cut axonal ends into functional and aberrant growth cones. J Comp Neurol 507:spc1Google Scholar
  15. 15.
    Falnikar A, Baas PW (2009) Critical roles for microtubules in axonal development and disease. Results Probl Cell Differ. doi:10.1007/400_2009_2
  16. 16.
    Fazio R, Quattrini A, Bolognesi A et al (1999) Docetaxel neuropathy: a distal axonopathy. Acta Neuropathol 98:651–653CrossRefPubMedGoogle Scholar
  17. 17.
    Flatters SJ, Bennett GJ (2006) Studies of peripheral sensory nerves in paclitaxel-induced painful peripheral neuropathy: evidence for mitochondrial dysfunction. Pain 122:245–257CrossRefPubMedGoogle Scholar
  18. 18.
    Foss M, Wilcox BW, Alsop GB, Zhang D (2008) Taxol crystals can masquerade as stabilized microtubules. PLoS ONE 3:e1476CrossRefPubMedGoogle Scholar
  19. 19.
    Galjart N (2005) CLIPs and CLASPs and cellular dynamics. Nat Rev Mol Cell Biol 6:487–498CrossRefPubMedGoogle Scholar
  20. 20.
    Gitler D, Spira ME (1998) Real time imaging of calcium-induced localized proteolytic activity after axotomy and its relation to growth cone formation. Neuron 20:1123–1135CrossRefPubMedGoogle Scholar
  21. 21.
    Hayashi K, Kawai-Hirai R, Ishikawa K, Takata K (2002) Reversal of neuronal polarity characterized by conversion of dendrites into axons in neonatal rat cortical neurons in vitro. Neuroscience 110:7–17CrossRefPubMedGoogle Scholar
  22. 22.
    Hendricks M, Jesuthasan S (2009) PHR regulates growth cone pausing at intermediate targets through microtubule disassembly. J Neurosci 29:6593–6598CrossRefPubMedGoogle Scholar
  23. 23.
    Jaworski J, Hoogenraad CC, Akhmanova A (2008) Microtubule plus-end tracking proteins in differentiated mammalian cells. Int J Biochem Cell Biol 40:619–637CrossRefPubMedGoogle Scholar
  24. 24.
    Jaworski J, Kapitein LC, Gouveia SM et al (2009) Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron 61:85–100CrossRefPubMedGoogle Scholar
  25. 25.
    Jordan MA, Kamath K (2007) How do microtubule-targeted drugs work? An overview. Curr Cancer Drug Targets 7:730–742CrossRefPubMedGoogle Scholar
  26. 26.
    Jordan MA, Wendell K, Gardiner S et al (1996) Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death. Cancer Res 56:816–825PubMedGoogle Scholar
  27. 27.
    Kar S, Fan J, Smith MJ, Goedert M, Amos LA (2003) Repeat motifs of tau bind to the insides of microtubules in the absence of taxol. Embo J 22:70–77CrossRefPubMedGoogle Scholar
  28. 28.
    Kim T, Chang S (2006) Quantitative evaluation of the mode of microtubule transport in Xenopus neurons. Mol Cells 21:76–81PubMedGoogle Scholar
  29. 29.
    Komarova Y, De Groot CO, Grigoriev I et al (2009) Mammalian end binding proteins control persistent microtubule growth. J Cell Biol 184:691–706CrossRefPubMedGoogle Scholar
  30. 30.
    Lee JJ, Swain SM (2006) Peripheral neuropathy induced by microtubule-stabilizing agents. J Clin Oncol 24:1633–1642CrossRefPubMedGoogle Scholar
  31. 31.
    Letourneau PC, Ressler AH (1984) Inhibition of neurite initiation and growth by taxol. J Cell Biol 98:1355–1362CrossRefPubMedGoogle Scholar
  32. 32.
    Lewcock JW, Genoud N, Lettieri K, Pfaff SL (2007) The ubiquitin ligase Phr1 regulates axon outgrowth through modulation of microtubule dynamics. Neuron 56:604–620CrossRefPubMedGoogle Scholar
  33. 33.
    Lodish H, Berk A, Zipursky L et al (2000) Molecular cell biology, 4th edn. W. H. Freeman, New YorkGoogle Scholar
  34. 34.
    Ma Y, Shakiryanova D, Vardya I, Popov SV (2004) Quantitative analysis of microtubule transport in growing nerve processes. Curr Biol 14:725–730CrossRefPubMedGoogle Scholar
  35. 35.
    Masurovsky EB, Peterson ER, Crain SM, Horwitz SB (1981) Microtubule arrays in taxol-treated mouse dorsal root ganglion–spinal cord cultures. Brain Res 217:392–398CrossRefPubMedGoogle Scholar
  36. 36.
    Mielke S, Sparreboom A, Mross K (2006) Peripheral neuropathy: a persisting challenge in paclitaxel-based regimes. Eur J Cancer 42:24–30CrossRefPubMedGoogle Scholar
  37. 37.
    Nakagawa H, Koyama K, Murata Y et al (2000) EB3, a novel member of the EB1 family preferentially expressed in the central nervous system, binds to a CNS-specific APC homologue. Oncogene 19:210–216CrossRefPubMedGoogle Scholar
  38. 38.
    Nakata T, Yorifuji H (1999) Morphological evidence of the inhibitory effect of taxol on the fast axonal transport. Neurosci Res 35:113–122CrossRefPubMedGoogle Scholar
  39. 39.
    Perez F, Diamantopoulos GS, Stalder R, Kreis TE (1999) CLIP-170 highlights growing microtubule ends in vivo. Cell 96:517–527CrossRefPubMedGoogle Scholar
  40. 40.
    Pignata S, De Placido S, Biamonte R et al (2006) Residual neurotoxicity in ovarian cancer patients in clinical remission after first-line chemotherapy with carboplatin and paclitaxel: the Multicenter Italian Trial in Ovarian cancer (MITO-4) retrospective study. BMC Cancer 6:5CrossRefPubMedGoogle Scholar
  41. 41.
    Postma TJ, Vermorken JB, Liefting AJ, Pinedo HM, Heimans JJ (1995) Paclitaxel-induced neuropathy. Ann Oncol 6:489–494PubMedGoogle Scholar
  42. 42.
    Rajnicek AM, Foubister LE, McCaig CD (2006) Growth cone steering by a physiological electric field requires dynamic microtubules, microfilaments and Rac-mediated filopodial asymmetry. J Cell Sci 119:1736–1745CrossRefPubMedGoogle Scholar
  43. 43.
    Ruthel G, Hollenbeck PJ (2003) Response of mitochondrial traffic to axon determination and differential branch growth. J Neurosci 23:8618–8624PubMedGoogle Scholar
  44. 44.
    Sahenk Z, Barohn R, New P, Mendell JR (1994) Taxol neuropathy. Electrodiagnostic and sural nerve biopsy findings. Arch Neurol 51:726–729PubMedGoogle Scholar
  45. 45.
    Sahly I, Erez H, Khoutorsky A, Shapira E, Spira ME (2003) Effective expression of the green fluorescent fusion proteins in cultured Aplysia neurons. J Neurosci Methods 126:111–117CrossRefPubMedGoogle Scholar
  46. 46.
    Sahly I, Khoutorsky A, Erez H, Prager-Khoutorsky M, Spira ME (2006) On-line confocal imaging of the events leading to structural dedifferentiation of an axonal segment into a growth cone after axotomy. J Comp Neurol 494:705–720CrossRefPubMedGoogle Scholar
  47. 47.
    Scuteri A, Nicolini G, Miloso M et al (2006) Paclitaxel toxicity in post-mitotic dorsal root ganglion (DRG) cells. Anticancer Res 26:1065–1070PubMedGoogle Scholar
  48. 48.
    Seitz A, Kojima H, Oiwa K et al (2002) Single-molecule investigation of the interference between kinesin, tau and MAP2c. Embo J 21:4896–4905CrossRefPubMedGoogle Scholar
  49. 49.
    Shemesh OA, Erez H, Ginzburg I, Spira ME (2008) Tau-induced traffic jams reflect organelles accumulation at points of microtubule polar mismatching. Traffic 9:458–471CrossRefPubMedGoogle Scholar
  50. 50.
    Siau C, Xiao W, Bennett GJ (2006) Paclitaxel- and vincristine-evoked painful peripheral neuropathies: loss of epidermal innervation and activation of Langerhans cells. Exp Neurol 201:507–514CrossRefPubMedGoogle Scholar
  51. 51.
    Spira ME, Dormann A, Ashery U et al (1996) Use of Aplysia neurons for the study of cellular alterations and the resealing of transected axons in vitro. J Neurosci Methods 69:91–102CrossRefPubMedGoogle Scholar
  52. 52.
    Stepanova T, Slemmer J, Hoogenraad CC et al (2003) Visualization of microtubule growth in cultured neurons via the use of EB3-GFP (end-binding protein 3-green fluorescent protein). J Neurosci 23:2655–2664PubMedGoogle Scholar
  53. 53.
    Takahashi D, Yu W, Baas PW, Kawai-Hirai R, Hayashi K (2007) Rearrangement of microtubule polarity orientation during conversion of dendrites to axons in cultured pyramidal neurons. Cell Motil Cytoskeleton 64:347–359CrossRefPubMedGoogle Scholar
  54. 54.
    Theiss C, Meller K (2000) Taxol impairs anterograde axonal transport of microinjected horseradish peroxidase in dorsal root ganglia neurons in vitro. Cell Tissue Res 299:213–224CrossRefPubMedGoogle Scholar
  55. 55.
    Verstreken P, Ly CV, Venken KJ et al (2005) Synaptic mitochondria are critical for mobilization of reserve pool vesicles at Drosophila neuromuscular junctions. Neuron 47:365–378CrossRefPubMedGoogle Scholar
  56. 56.
    Williamson T, Gordon-Weeks PR, Schachner M, Taylor J (1996) Microtubule reorganization is obligatory for growth cone turning. Proc Natl Acad Sci USA 93:15221–15226CrossRefPubMedGoogle Scholar
  57. 57.
    Winer EP, Berry DA, Woolf S et al (2004) Failure of higher-dose paclitaxel to improve outcome in patients with metastatic breast cancer: cancer and leukemia group B trial 9342. J Clin Oncol 22:2061–2068CrossRefPubMedGoogle Scholar
  58. 58.
    Xiao WH, Bennett GJ (2008) C-fiber spontaneous discharge evoked by chronic inflammation is suppressed by a long-term infusion of lidocaine yielding nanogram per milliliter plasma levels. Pain 137:218–228CrossRefPubMedGoogle Scholar
  59. 59.
    Xiao WH, Bennett GJ (2008) Chemotherapy-evoked neuropathic pain: abnormal spontaneous discharge in A-fiber and C-fiber primary afferent neurons and its suppression by acetyl-l-carnitine. Pain 135:262–270CrossRefPubMedGoogle Scholar
  60. 60.
    Yu W, Cook C, Sauter C et al (2000) Depletion of a microtubule-associated motor protein induces the loss of dendritic identity. J Neurosci 20:5782–5791PubMedGoogle Scholar
  61. 61.
    Zhang C, Sriratana A, Minamikawa T, Nagley P (1998) Photosensitisation properties of mitochondrially localised green fluorescent protein. Biochem Biophys Res Commun 242:390–395CrossRefPubMedGoogle Scholar
  62. 62.
    Zheng Y, Wildonger J, Ye B et al (2008) Dynein is required for polarized dendritic transport and uniform microtubule orientation in axons. Nat Cell Biol 10:1172–1180CrossRefPubMedGoogle Scholar
  63. 63.
    Ziv NE, Spira ME (1997) Localized and transient elevations of intracellular Ca2+ induce the dedifferentiation of axonal segments into growth cones. J Neurosci 17:3568–3579PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of Neurobiology, Institute of Life ScienceThe Hebrew University of JerusalemJerusalemIsrael

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