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Axon Viability and Mitochondrial Function are Dependent on Local Protein Synthesis in Sympathetic Neurons

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Abstract

(1) Axons contain numerous mRNAs and a local protein synthetic system that can be regulated independently of the cell body. (2) In this study, cultured primary sympathetic neurons were employed, to assess the effect of local protein synthesis blockade on axon viability and mitochondrial function. (3) Inhibition of local protein synthesis reduced newly synthesized axonal proteins by 65% and resulted in axon retraction after 6 h. Acute inhibition of local protein synthesis also resulted in a significant decrease in the membrane potential of axonal mitochondria. Likewise, blockade of local protein transport into the mitochondria by transfection of the axons with Hsp90 C-terminal domain decreased the mitochondrial membrane potential by 65%. Moreover, inhibition of the local protein synthetic system also reduced the ability of mitochondria to restore axonal levels of ATP after KCl-induced depolarization. (4) Taken together, these results indicate that the local protein synthetic system plays an important role in mitochondrial function and the maintenance of the axon.

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References

  • Anderson S, Bankier AT, Barrell BG, de Bruijn MHL, Coulson AR, Drouin J, Eperon IC, Nierlich DP, Roe BA, Sanger F, Schreier PH, Smith AJH, Staden R, Young IG (1981) Sequence and organization of the human mitochondrial genome. Nature 290:457–465

    Article  PubMed  CAS  Google Scholar 

  • Attardi G, Schatz G (1988) Biogenesis of mitochondria. Annu Rev Cell Biol 4:289–331

    Article  PubMed  CAS  Google Scholar 

  • Bauer MF, Hofmann S (2006) Import of mitochondrial proteins. In: Schapira AHV (ed) Mitochondrial function and dysfunction. Academic Press, San Diego, CA, pp 57–90

    Google Scholar 

  • Beaumont V, Zhong N, Fletcher R, Froemke RC, Zucker RS (2001) Phosphorylation and local presynaptic protein synthesis in calcium- and calcineurin-dependent induction of crayfish long-term facilitation. Neuron 32:489–501

    Article  PubMed  CAS  Google Scholar 

  • Bleher R, Martin R (2001) Ribosomes in the squid giant axon. Neuroscience 107:527–534

    Article  PubMed  CAS  Google Scholar 

  • Brittis PA, Lu Q, Flanagan JG (2002) Axonal protein synthesis provides a mechanism for localized regulation at an intermediate target. Cell 110:223–235

    Article  PubMed  CAS  Google Scholar 

  • Campbell DS, Holt CE (2001) Chemotropic responses of retinal growth cones mediated by rapid local protein synthesis and degradation. Neuron 32:1013–1026

    Article  PubMed  CAS  Google Scholar 

  • Campenot RB, Martin G (2001) Construction and use of compartmented cultures for studies of cell biology in neurons. In: Federoff S, Richadrson A (ed) Protocols for neural cell culture. Humana Press, Inc., Totowa, NJ, pp 49–57

    Chapter  Google Scholar 

  • Casadio A, Martin KC, Giustetto M, Zhu H, Chen M, Bartsch D, Bailey CH, Kandel ER (1999) A transient, neuron-wide form of CREB-mediated long-term facilitation can be stabilized at specific synapses by local protein synthesis. Cell 99:221–237

    Article  PubMed  CAS  Google Scholar 

  • Cossarizza A, Baccaranicontri M, Kalashnikova G, Franceschi C (1993) A new method for the cytofluorometric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide (JC-1). Biochem Biophys Res Commun 197:40–45

    Article  PubMed  CAS  Google Scholar 

  • Ehrenberg B, Montana V, Wei MD, Wuskell JP, Loew LM (1988) Membrane potential can be determined in individual cells from the nernstian distribution of cationic dyes. Biophys J 53:785–794

    Article  PubMed  CAS  Google Scholar 

  • Eng H, Lund K, Campenot RB (1999) Synthesis of beta-tubulin, actin, and other proteins in axons of sympathetic neurons in compartmented cultures. J Neurosci 19:1–9

    PubMed  CAS  Google Scholar 

  • Gioio AE, Eyman M, Zhang H, Lavina ZS, Giuditta A, Kaplan BB (2001) Local synthesis of nuclear-encoded mitochondrial proteins in the presynaptic nerve terminal. J Neurosci Res 64:447–453

    Article  PubMed  CAS  Google Scholar 

  • Gioio AE, Lavina ZS, Jurkovicova D, Zhang H, Eyman M, Giuditta A, Kaplan BB (2004) Nerve terminals of squid photoreceptor neurons contain a heterogeneous population of mRNAs and translate a transfected reporter mRNA. Eur J Neurosci 20:865–872

    Article  PubMed  Google Scholar 

  • Giuditta A, Kaplan BB, van Minnen J, Alvarez J, Koenig E (2002) Axonal and presynaptic protein synthesis: new insights into the biology of the neuron. Trends Neurosci 25:400–404

    Article  PubMed  CAS  Google Scholar 

  • Giuditta A, Menichini E, Perrone Capano C, Langella M, Martin R, Castigli E, Kaplan BB (1991) Active polysomes in the axoplasm of the squid giant axon. J Neurosci Res 28:18–28

    Article  PubMed  CAS  Google Scholar 

  • Hanz S, Perlson E, Willis D, Zheng JQ, Massarwa R, Huerta JJ, Koltzenburg M, Kohler M, van-Minnen J, Twiss JL, Fainzilber M (2003) Axoplasmic importins enable retrograde injury signaling in lesioned nerve. Neuron 40:1095–1104

    Article  PubMed  CAS  Google Scholar 

  • Heales SJR, Gegg ME, Clark JB (2006) Oxidative phosporylation: structure, function, and intermediary metabolism. In: Schapira AHV (ed) Mitochondria function and dysfuntion. Academic Press, San Diego, CA, pp 25–56

    Google Scholar 

  • Hollenbeck PJ, Saxton WM (2005) The axonal transport of mitochondria. J Cell Sci 118:5411–5419

    Article  PubMed  CAS  Google Scholar 

  • Johnson MI (2001) Primary cultures of sympathetic ganglia. In: Federoff S, Richadrson A (eds) Protocols for neural cell culture. Humana Press, Inc., Totowa, NJ, pp 71–94

    Chapter  Google Scholar 

  • Kaplan BB, Gioio AE, Perrone Capano C, Crispino M, Giuditta A (1992) β-Actin and β-Tubulin are components of a heterogeneous mRNA population present in the squid giant axon. Mol Cell Neurosci 3:133–144

    Article  CAS  Google Scholar 

  • Lee SK, Hollenbeck PJ (2003) Organization and translation of mRNA in sympathetic axons. J Cell Sci 116:4467–4478

    Article  PubMed  CAS  Google Scholar 

  • Li C, Sasaki Y, Takei K, Yamamoto H, Shouji M, Sugiyama Y, Kawakami T, Nakamura F, Yagi T, Ohshima T, Goshima Y (2004) Correlation between Semaphorin3A-induced facilitation of axonal transport and local activation of a translation initiation factor eukaryotic translation initiation factor 4E. J Neurosci 24:6161–6170

    Article  PubMed  CAS  Google Scholar 

  • Li J-Y, Volknandt W, Dahlstrom A, Herrmann C, Blasi J, Das B, Zimmermann H (1999) Axonal transport of ribonucleoprotein particles (Vaults). Neuroscience 91:1055–1065

    Article  PubMed  CAS  Google Scholar 

  • Liu K, Hu J-Y, Wang D, Schacher S (2003) Protein synthesis at synapse versus cell body: enhanced but transient expression of long-term facilitation at isolated synapses. J Neurobiol 56:275–286

    Article  PubMed  CAS  Google Scholar 

  • Liu S, Wong-Riley M (1994) Nuclear-encoded mitochondrial precursor protein: intramitochondrial delivery to dendrites and axon terminals of neurons and regulation by neuronal activity. J Neurosci 14:5338–5351

    PubMed  CAS  Google Scholar 

  • Margeot A, Garcia M, Wang W, Tetaud E, di Rago JP, Jacq C (2005) Why are many mRNAs translated to the vicinity of mitochondria: a role in protein complex assembly? Gene 354:64–71

    Article  PubMed  CAS  Google Scholar 

  • Martin KC, Barad M, Kandel ER (2000) Local protein synthesis and its role in synapse-specific plasticity. Curr Opin Neurobiol 10:587–592

    Article  PubMed  CAS  Google Scholar 

  • Martin KC, Casadio A, Zhu H, E Y, Rose JC, Chen M, Bailey CH, Kandel ER (1997) Synapse-specific, long-term facilitation of Aplysia sensory to motor synapses: a function for local protein synthesis in memory storage. Cell 91:927–938

    Article  PubMed  CAS  Google Scholar 

  • Ming GL, Wong ST, Henley J, Yuan XB, Song HJ, Spitzer NC, Poo M (2002) Adaptation in the chemotactic guidance of nerve growth cones. Nature 417:411–418

    Article  PubMed  CAS  Google Scholar 

  • Moccia R, Chen D, Lyles V, Kapuya E, E Y, Kalachikov S, Spahn CMT, Frank J, Kandel ER, Barad M, Martin KC (2003) An unbiased cDNA library prepared from isolated Aplysia sensory neuron processes is enriched for cytoskeletal and translational mRNAs. J Neurosci 23:9409–9417

    PubMed  CAS  Google Scholar 

  • Olink-Coux M, Hollenbeck PJ (1996) Localization and active transport of mRNA in axons of sympathetic neurons in culture. J Neurosci 16:1346–1358

    PubMed  CAS  Google Scholar 

  • Perrone Capano C, Giuditta A, Castigli E, Kaplan BB (1987) Occurrence and sequence complexity of polyadenylated RNA in squid axoplasm. J Neurochem 49:698–704

    Article  CAS  Google Scholar 

  • Piper M, Holt C (2004) RNA translations in axons. Annu Rev Cell Dev Biol 20:505–523

    Article  PubMed  CAS  Google Scholar 

  • Schacher S, Wu F (2002) Synapse formation in the absence of cell bodies requires protein synthesis. J Neurosci 22:1831–1839

    PubMed  CAS  Google Scholar 

  • Schuman EM, Dynes JL, Steward O (2006) Synaptic regulation of translation of dendritic mRNAs. J Neurosci 26:7143–7146

    Article  PubMed  CAS  Google Scholar 

  • Si K, Giustetto M, Etkin A, Hsu R, Janisiewicz AM, Miniaci MC, Kim JH, Zhu H, Kandel ER (2003) A neuronal isoform of CPEB regulates local protein synthesis and stabilizes synapse-specific long-term facilitation in Aplysia. Cell 115:893–904

    Article  PubMed  CAS  Google Scholar 

  • Truscott KN, Brandner K, Pfanner N (2003) Mechanisms of protein import into mitochondria. Curr Biol 13:R326–R337

    Article  PubMed  CAS  Google Scholar 

  • Young JC, Hoogenraad NJ, Hartl FU (2003) Molecular chaperones Hsp90 and Hsp70 deliver preproteins to the mitochondrial import receptor Tom70. Cell 112:41–50

    Article  PubMed  CAS  Google Scholar 

  • Young JC, Obermann WM, Hartl FU (1998) Specific binding of tetratricopeptide repeat proteins to the C-terminal 12-kDa domain of Hsp90. J Biol Chem 273:18007–18010

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Poo M (2002) Localized synaptic potentiation by BDNF requires local protein synthesis in the developing axon. Neuron 36:675–688

    Article  PubMed  CAS  Google Scholar 

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Acknowdgement

We thank Dr. J. Young (McGill Faculty of Medicine, Montreal, Quebec, Canada) for providing the C90 plasmid.

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Correspondence to Barry B. Kaplan.

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Hillefors, M., Gioio, A.E., Mameza, M.G. et al. Axon Viability and Mitochondrial Function are Dependent on Local Protein Synthesis in Sympathetic Neurons. Cell Mol Neurobiol 27, 701–716 (2007). https://doi.org/10.1007/s10571-007-9148-y

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  • DOI: https://doi.org/10.1007/s10571-007-9148-y

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