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The Journal of Neuroscience, January 1, 1999, 19(1):1-9

Synthesis of beta -Tubulin, Actin, and Other Proteins in Axons of Sympathetic Neurons in Compartmented Cultures

Hubert Eng, Karen Lund, and Robert B. Campenot

Department of Cell Biology, University of Alberta, Edmonton, Alberta, T6G 2H7 Canada

The proteins needed for growth and maintenance of the axon are generally believed to be synthesized in the cell bodies and delivered to the axons by anterograde transport. However, recent reports suggest that some proteins can also be synthesized within axons. We used [35S]methionine metabolic labeling to investigate axonal protein synthesis in compartmented cultures of sympathetic neurons from newborn rats. Incubation of distal axons for 4 hr with [35S]methionine resulted in a highly specific pattern of labeled axonal proteins on SDS-PAGE, with 4 prominent bands in the 43-55 kDa range. The labeled proteins in axons were not synthesized in the cell bodies, because they were also produced by axons after the cell bodies had been removed. Two of the proteins were identified by immunoprecipitation as actin and beta -tubulin. Axons synthesized <1% of the actin and tubulin synthesized in the cell bodies and transported into the axons, and 75-85% inhibition of axonal protein synthesis by cycloheximide and puromycin failed to inhibit axonal elongation. Nonetheless, the specific production by axons of the major proteins of the axonal cytoskeleton suggests that axonal protein synthesis arises from specific mechanisms and likely has biological significance. One hypothetical scenario involves neurons with long axons in vivo in which losses from turnover during axonal transport may limit the availability of cell body synthesized proteins to the distal axons. In this case, a significant fraction of axonal proteins might be supplied by axonal synthesis, which could, therefore, play important roles in axonal maintenance, regeneration, and sprouting.

Key words: actin; tubulin; axonal protein synthesis; compartmented cultures; [35S]methionine metabolic labeling; axon growth


Copyright © 1999 Society for Neuroscience  0270-6474/99/1911-09$05.00/0


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A. Jovanovska and R. A. Prosser
Translational and Transcriptional Inhibitors Block Serotonergic Phase Advances of the Suprachiasmatic Nucleus Circadian Pacemaker In Vitro
J Biol Rhythms, April 1, 2002; 17(2): 137 - 146.
[Abstract] [PDF]


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J. Physiol.Home page
M. J Nicol and B. Walmsley
Ultrastructural basis of synaptic transmission between endbulbs of Held and bushy cells in the rat cochlear nucleus
J. Physiol., March 15, 2002; 539(3): 713 - 723.
[Abstract] [Full Text] [PDF]


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J. Neurophysiol.Home page
G. Losi, K. Prybylowski, Z. Fu, J. H. Luo, and S. Vicini
Silent Synapses in Developing Cerebellar Granule Neurons
J Neurophysiol, March 1, 2002; 87(3): 1263 - 1270.
[Abstract] [Full Text] [PDF]


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J. Neurophysiol.Home page
D. M. Senseman and K. A. Robbins
High-Speed VSD Imaging of Visually Evoked Cortical Waves: Decomposition Into Intra- and Intercortical Wave Motions
J Neurophysiol, March 1, 2002; 87(3): 1499 - 1514.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
T. Tsunemi, H. Saegusa, K. Ishikawa, S. Nagayama, T. Murakoshi, H. Mizusawa, and T. Tanabe
Novel Cav2.1 Splice Variants Isolated from Purkinje Cells Do Not Generate P-type Ca2+ Current
J. Biol. Chem., February 22, 2002; 277(9): 7214 - 7221.
[Abstract] [Full Text] [PDF]


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Cereb CortexHome page
B. Roerig and B. Chen
Relationships of Local Inhibitory and Excitatory Circuits to Orientation Preference Maps in Ferret Visual Cortex
Cereb Cortex, February 1, 2002; 12(2): 187 - 198.
[Abstract] [Full Text] [PDF]


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BrainHome page
C. Stadelmann, M. Kerschensteiner, T. Misgeld, W. Bruck, R. Hohlfeld, and H. Lassmann
BDNF and gp145trkB in multiple sclerosis brain lesions: neuroprotective interactions between immune and neuronal cells?
Brain, January 1, 2002; 125(1): 75 - 85.
[Abstract] [Full Text] [PDF]


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J. Neurophysiol.Home page
C. Gonzalez-Islas and J. J. Hablitz
Dopamine Inhibition of Evoked IPSCs in Rat Prefrontal Cortex
J Neurophysiol, December 1, 2001; 86(6): 2911 - 2918.
[Abstract] [Full Text] [PDF]


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NeuroscientistHome page
E. L. Meyer-Bernstein and A. Sehgal
Book Review: Molecular Regulation of Circadian Rhythms in Drosophila and Mammals
Neuroscientist, December 1, 2001; 7(6): 496 - 505.
[Abstract] [PDF]


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J. Neurosci.Home page
J.-Q. Zheng, T. K. Kelly, B. Chang, S. Ryazantsev, A. K. Rajasekaran, K. C. Martin, and J. L. Twiss
A Functional Role for Intra-Axonal Protein Synthesis during Axonal Regeneration from Adult Sensory Neurons
J. Neurosci., December 1, 2001; 21(23): 9291 - 9303.
[Abstract] [Full Text] [PDF]


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HypertensionHome page
K. Matsumura, T. Tsuchihashi, and I. Abe
Central Human Cocaine- and Amphetamine-Regulated Transcript Peptide 55-102 Increases Arterial Pressure in Conscious Rabbits
Hypertension, November 1, 2001; 38(5): 1096 - 1100.
[Abstract] [Full Text] [PDF]


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Genes Dev.Home page
Y.-N. Jan and L. Y. Jan
Dendrites
Genes & Dev., October 15, 2001; 15(20): 2627 - 2641.
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J. Physiol.Home page
J. Reisert and H. R Matthews
Simultaneous recording of receptor current and intraciliary Ca2+ concentration in salamander olfactory receptor cells
J. Physiol., September 15, 2001; 535(3): 637 - 645.
[Abstract] [Full Text] [PDF]


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J. Neurophysiol.Home page
P. A. Gusev and D. L. Alkon
Intracellular Correlates of Spatial Memory Acquisition in Hippocampal Slices: Long-Term Disinhibition of CA1 Pyramidal Cells
J Neurophysiol, August 1, 2001; 86(2): 881 - 899.
[Abstract] [Full Text] [PDF]


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J Biol RhythmsHome page
W. J. Schwartz, H. O. de la Iglesia, P. Zlomanczuk, and H. Illnerova
Encoding Le Quattro Stagioni within the Mammalian Brain: Photoperiodic Orchestration through the Suprachiasmatic Nucleus
J Biol Rhythms, August 1, 2001; 16(4): 302 - 311.
[Abstract] [PDF]


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EndocrinologyHome page
C. R. Abbott, M. Rossi, A. M. Wren, K. G. Murphy, A. R. Kennedy, S. A. Stanley, A. N. Zollner, D. G. A. Morgan, I. Morgan, M. A. Ghatei, et al.
Evidence of an Orexigenic Role for Cocaine- and Amphetamine-Regulated Transcript after Administration into Discrete Hypothalamic Nuclei
Endocrinology, August 1, 2001; 142(8): 3457 - 3463.
[Abstract] [Full Text] [PDF]


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Cereb CortexHome page
Z. F. Kisvarday, P. Buzas, and U. T. Eysel
Calculating Direction Maps from Intrinsic Signals revealed by Optical Imaging
Cereb Cortex, July 1, 2001; 11(7): 636 - 647.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
G. S. Pollock, E. Vernon, M. E. Forbes, Q. Yan, Y.-T. Ma, T. Hsieh, R. Robichon, D. O. Frost, and J. E. Johnson
Effects of Early Visual Experience and Diurnal Rhythms on BDNF mRNA and Protein Levels in the Visual System, Hippocampus, and Cerebellum
J. Neurosci., June 1, 2001; 21(11): 3923 - 3931.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
S. Aja, G. J. Schwartz, M. J. Kuhar, and T. H. Moran
Intracerebroventricular CART peptide reduces rat ingestive behavior and alters licking microstructure
Am J Physiol Regulatory Integrative Comp Physiol, June 1, 2001; 280(6): R1613 - R1619.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
D. Pfaff
Precision in mouse behavior genetics
PNAS, May 3, 2001; (2001) 101128598.
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Chem SensesHome page
K. A. Osborne, J. S. de Belle, and M. B. Sokolowski
Foraging Behaviour in Drosophila Larvae: Mushroom Body Ablation
Chem Senses, February 1, 2001; 26(2): 223 - 230.
[Abstract] [Full Text] [PDF]


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Mol. Pharmacol.Home page
R. D. Mayfield and N. R. Zahniser
Dopamine D2 Receptor Regulation of the Dopamine Transporter Expressed in Xenopus laevis Oocytes Is Voltage-Independent
Mol. Pharmacol., January 1, 2001; 59(1): 113 - 121.
[Abstract] [Full Text]


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J. Physiol.Home page
J. Reisert and H. R Matthews
Response properties of isolated mouse olfactory receptor cells
J. Physiol., January 1, 2001; 530(1): 113 - 122.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
T. R. Cummins, J. A. Black, S. D. Dib-Hajj, and S. G. Waxman
Glial-Derived Neurotrophic Factor Upregulates Expression of Functional SNS and NaN Sodium Channels and Their Currents in Axotomized Dorsal Root Ganglion Neurons
J. Neurosci., December 1, 2000; 20(23): 8754 - 8761.
[Abstract] [Full Text] [PDF]


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J. Physiol.Home page
D. Ragozzino, A. Giovannelli, V. Degasperi, F. Eusebi, and F. Grassi
Zinc permeates mouse muscle ACh receptor channels expressed in BOSC 23 cells and affects channel function
J. Physiol., November 15, 2000; 529(1): 83 - 91.
[Abstract] [Full Text] [PDF]


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JGPHome page
N. Zilberberg, N. Ilan, R. Gonzalez-Colaso, and S. A.N. Goldstein
Opening and Closing of KcnkO Potassium Leak Channels Is Tightly Regulated
J. Gen. Physiol., November 1, 2000; 116(5): 721 - 734.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
T. Tamaru, Y. Isojima, T. Yamada, M. Okada, K. Nagai, and K. Takamatsu
Light and Glutamate-Induced Degradation of the Circadian Oscillating Protein BMAL1 during the Mammalian Clock Resetting
J. Neurosci., October 15, 2000; 20(20): 7525 - 7530.
[Abstract] [Full Text] [PDF]


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J. Neurosci.Home page
V. Suri, J. C. Hall, and M. Rosbash
Two Novel doubletime Mutants Alter Circadian Properties and Eliminate the Delay between RNA and Protein in Drosophila
J. Neurosci., October 15, 2000; 20(20): 7547 - 7555.
[Abstract] [Full Text] [PDF]


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J. Physiol.Home page
R A Shiells and G Falk
Activation of Ca2+-calmodulin kinase II induces desensitization by background light in dogfish retinal 'on' bipolar cells
J. Physiol., October 15, 2000; 528(2): 327 - 338.
[Abstract] [Full Text] [PDF]


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J Biol RhythmsHome page
S. M. Reppert and D. R. Weaver
Comparing Clockworks: Mouse versus Fly
J Biol Rhythms, October 1, 2000; 15(5): 357 - 364.
[PDF]


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J. Physiol.Home page
G. A Kerchner, L. M T Canzoniero, S. P. Yu, C. Ling, and D. W Choi
Zn2+ current is mediated by voltage-gated Ca2+ channels and enhanced by extracellular acidity in mouse cortical neurones
J. Physiol., October 1, 2000; 528(1): 39 - 52.
[Abstract] [Full Text] [PDF]



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