Tools for Analyzing the Role of Local Protein Synthesis in Synaptic Plasticity

Protocol
Part of the Neuromethods book series (NM, volume 105)

Abstract

Numerous mRNA transcripts are transported to dendrites for local protein translation, which may provide a mechanism for selectively and rapidly modulating synapses in response to stimulation. Indeed, highly specific activity-dependent neuronal responses are critical for modulating synapses in order to facilitate learning and memory. Brain-derived neurotrophic factor (BDNF) is a small, secreted protein that has been shown to play critical roles in synaptic plasticity. The gene for BDNF produces two pools of mRNA, with either a short or a long 3′ untranslated region (3′ UTR). Short 3′ UTR Bdnf mRNA is restricted to the soma, while long 3′ UTR Bdnf mRNA is localized to both the soma as well as dendrites for local translation. Recent in vivo and in vitro studies have demonstrated that dendritically synthesized BDNF, derived from long 3′ UTR Bdnf mRNA, is required for spine maturation and spine pruning. In this chapter, we describe methodology used to understand the molecular mechanisms through which dendritically synthesized proteins (i.e., BDNF) modulate synaptic plasticity.

Key words

Brain-derived neurotrophic factor (BDNF) 3′ UTR mRNA trafficking Local translation Dendritic spine In situ hybridization 
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References

  1. 1.
    Harris KM (1999) Structure, development, and plasticity of dendritic spines. Curr Opin Neurobiol 9:343–348CrossRefPubMedGoogle Scholar
  2. 2.
    Hering H, Sheng M (2001) Dendritic spines: structure, dynamics and regulation. Nat Rev Neurosci 2:880–888CrossRefPubMedGoogle Scholar
  3. 3.
    Calabrese B, Wilson MS, Halpain S (2006) Development and regulation of dendritic spine synapses. Physiology (Bethesda) 21:38–47CrossRefGoogle Scholar
  4. 4.
    Holtmaat A, Svoboda K (2009) Experience-dependent structural synaptic plasticity in the mammalian brain. Nat Rev Neurosci 10:647–658CrossRefPubMedGoogle Scholar
  5. 5.
    Yoshihara Y, De Roo M, Muller D (2009) Dendritic spine formation and stabilization. Curr Opin Neurobiol 19:146–153CrossRefPubMedGoogle Scholar
  6. 6.
    Bramham CR, Wells DG (2007) Dendritic mRNA: transport, translation and function. Nat Rev Neurosci 8:776–789CrossRefPubMedGoogle Scholar
  7. 7.
    Burgin KE, Waxham MN, Rickling S et al (1990) In situ hybridization histochemistry of Ca2+/calmodulin-dependent protein kinase in developing rat brain. J Neurosci 10:1788–1798PubMedGoogle Scholar
  8. 8.
    Ju W, Morishita W, Tsui J et al (2004) Activity-dependent regulation of dendritic synthesis and trafficking of AMPA receptors. Nat Neurosci 7:244–253CrossRefPubMedGoogle Scholar
  9. 9.
    Steward O, Levy WB (1982) Preferential localization of polyribosomes under the base of dendritic spines in granule cells of the dentate gyrus. J Neurosci 2:284–291PubMedGoogle Scholar
  10. 10.
    Sutton MA, Ito HT, Cressy P et al (2006) Miniature neurotransmission stabilizes synaptic function via tonic suppression of local dendritic protein synthesis. Cell 125:785–799CrossRefPubMedGoogle Scholar
  11. 11.
    Kang H, Schuman EM (1996) A requirement for local protein synthesis in neurotrophin-induced hippocampal synaptic plasticity. Science 273:1402–1406CrossRefPubMedGoogle Scholar
  12. 12.
    Huber KM, Kayser MS, Bear MF (2000) Role for rapid dendritic protein synthesis in hippocampal mGluR-dependent long-term depression. Science 288:1254–1257CrossRefPubMedGoogle Scholar
  13. 13.
    Martin KC, Casadio A, Zhu H et al (1997) Synapse-specific, long-term facilitation of aplysia sensory to motor synapses: a function for local protein synthesis in memory storage. Cell 91:927–938CrossRefPubMedGoogle Scholar
  14. 14.
    Wang DO, Kim SM, Zhao Y et al (2009) Synapse- and stimulus-specific local translation during long-term neuronal plasticity. Science 324:1536–1540PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Soule J, Messaoudi E, Bramham CR (2006) Brain-derived neurotrophic factor and control of synaptic consolidation in the adult brain. Biochem Soc Trans 34:600–604CrossRefPubMedGoogle Scholar
  16. 16.
    Lu B, Martinowich K (2008) Cell biology of BDNF and its relevance to schizophrenia. Novartis Found Symp 289:119–129, discussion 129–35, 193–5PubMedCentralCrossRefPubMedGoogle Scholar
  17. 17.
    Waterhouse EG, Xu B (2009) New insights into the role of brain-derived neurotrophic factor in synaptic plasticity. Mol Cell Neurosci 42:81–89PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Gooney M, Shaw K, Kelly A et al (2002) Long-term potentiation and spatial learning are associated with increased phosphorylation of TrkB and extracellular signal-regulated kinase (ERK) in the dentate gyrus: evidence for a role for brain-derived neurotrophic factor. Behav Neurosci 116:455–463CrossRefPubMedGoogle Scholar
  19. 19.
    Tang SJ, Reis G, Kang H et al (2002) A rapamycin-sensitive signaling pathway contributes to long-term synaptic plasticity in the hippocampus. Proc Natl Acad Sci U S A 99:467–472PubMedCentralCrossRefPubMedGoogle Scholar
  20. 20.
    Kanhema T, Dagestad G, Panja D et al (2006) Dual regulation of translation initiation and peptide chain elongation during BDNF-induced LTP in vivo: evidence for compartment-specific translation control. J Neurochem 99:1328–1337CrossRefPubMedGoogle Scholar
  21. 21.
    Gomez-Palacio-Schjetnan A, Escobar ML (2008) In vivo BDNF modulation of adult functional and morphological synaptic plasticity at hippocampal mossy fibers. Neurosci Lett 445:62CrossRefPubMedGoogle Scholar
  22. 22.
    Lou H, Kim SK, Zaitsev E et al (2005) Sorting and activity-dependent secretion of BDNF require interaction of a specific motif with the sorting receptor carboxypeptidase e. Neuron 45:245–255CrossRefPubMedGoogle Scholar
  23. 23.
    An JJ, Gharami K, Liao GY et al (2008) Distinct role of long 3′ UTR BDNF mRNA in spine morphology and synaptic plasticity in hippocampal neurons. Cell 134:175–187PubMedCentralCrossRefPubMedGoogle Scholar
  24. 24.
    Waterhouse EG, An JJ, Orefice LL et al (2012) BDNF promotes differentiation and maturation of adult-born neurons through GABAergic transmission. J Neurosci 32:14318–14330PubMedCentralCrossRefPubMedGoogle Scholar
  25. 25.
    Orefice LL, Waterhouse EG, Partridge JG et al (2013) Distinct roles for somatically and dendritically synthesized brain-derived neurotrophic factor in morphogenesis of dendritic spines. J Neurosci 33:11618–11632PubMedCentralCrossRefPubMedGoogle Scholar
  26. 26.
    Timmusk T, Palm K, Metsis M et al (1993) Multiple promoters direct tissue-specific expression of the rat BDNF gene. Neuron 10:475–489CrossRefPubMedGoogle Scholar
  27. 27.
    Weickert CS, Hyde TM, Lipska BK et al (2003) Reduced brain-derived neurotrophic factor in prefrontal cortex of patients with schizophrenia. Mol Psychiatry 8:592–610CrossRefPubMedGoogle Scholar
  28. 28.
    Fischer M, Kaech S, Knutti D et al (1998) Rapid actin-based plasticity in dendritic spines. Neuron 20:847–854CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of NeuroscienceThe Scripps Research Institute FloridaJupiterUSA
  2. 2.Department of BiologyGeorgetown UniversityWashingtonUSA

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