Skip to main content
SpringerLink
Log in

BDNF Function in Long-Term Synaptic Plasticity in the Dentate Gyrus In Vivo: Methods for Local Drug Delivery and Biochemical Analysis of Translation

  • Protocol
  • First Online:
Brain-Derived Neurotrophic Factor (BDNF)

Part of the book series: Neuromethods ((NM,volume 143))

Abstract

Neurotrophins are essential for multiple aspects of neuronal development and to important functions like synaptic plasticity. Brain-derived neurotrophic factor (BDNF) is a critical activity-dependent modulator of gene expression which regulates both transcription and translation. BDNF is crucial in the maintenance of long-term potentiation (LTP) at synapses and regulates protein synthesis at the dendritic and synaptic level. To elucidate the mechanisms operating in the hippocampal dentate gyrus region, in vivo electrophysiology and pharmacology is combined with analysis of signaling pathways and protein synthesis. Here, we present methods for the analysis of translation initiation, polysome formation, and translational efficiency in the context of LTP consolidation in live rodents.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Bramham CR, Messaoudi E (2005) BDNF function in adult synaptic plasticity: the synaptic consolidation hypothesis. Prog Neurobiol 76:99–125

    Article  CAS  Google Scholar 

  2. Panja D, Bramham CR (2014) BDNF mechanisms in late LTP formation: a synthesis and breakdown. Neuropharmacology 76(Pt C):664–676

    Article  CAS  Google Scholar 

  3. Park H, Poo MM (2013) Neurotrophin regulation of neural circuit development and function. Nat Rev Neurosci 14:7–23

    Article  CAS  Google Scholar 

  4. Berninger B, Poo M (1996) Fast actions of neurotrophic factors. Curr Opin Neurobiol 6:324–330

    Article  CAS  Google Scholar 

  5. Lu B, Figurov A (1997) Role of neurotrophins in synapse development and plasticity. Rev Neurosci 8:1–12

    Article  CAS  Google Scholar 

  6. Thoenen H (1995) Neurotrophins and neuronal plasticity. Science 270:593–598

    Article  CAS  Google Scholar 

  7. Kang H, Schuman EM (1996) A requirement for local protein synthesis in neurotrophin-induced hippocampal synaptic plasticity. Science 273:1402–1406

    Article  CAS  Google Scholar 

  8. Liao L, Pilotte J, Xu T, Wong CC, Edelman GM, Vanderklish P, Yates JR 3rd (2007) BDNF induces widespread changes in synaptic protein content and up-regulates components of the translation machinery: an analysis using high-throughput proteomics. J Proteome Res 6:1059–1071

    Article  CAS  Google Scholar 

  9. Malenka RC, Bear MF (2004) LTP and LTD: an embarrassment of riches. Neuron 44:5–21

    Article  CAS  Google Scholar 

  10. Bliss TV, Collingridge GL (1993) A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361:31–39

    Article  CAS  Google Scholar 

  11. Frey U, Krug M, Reymann KG, Matthies H (1988) Anisomycin, an inhibitor of protein synthesis, blocks late phases of LTP phenomena in the hippocampal CA1 region in vitro. Brain Res 452:57–65

    Article  CAS  Google Scholar 

  12. Lynch MA (2004) Long-term potentiation and memory. Physiol Rev 84:87–136

    Article  CAS  Google Scholar 

  13. Otani S, Marshall CJ, Tate WP, Goddard GV, Abraham WC (1989) Maintenance of long-term potentiation in rat dentate gyrus requires protein synthesis but not messenger RNA synthesis immediately post-tetanization. Neuroscience 28:519–526

    Article  CAS  Google Scholar 

  14. Stanton PK, Sarvey JM (1984) Blockade of long-term potentiation in rat hippocampal CA1 region by inhibitors of protein synthesis. J Neurosci 4:3080–3088

    Article  CAS  Google Scholar 

  15. Lu Y, Christian K, Lu B (2008) BDNF: a key regulator for protein synthesis-dependent LTP and long-term memory? Neurobiol Learn Mem 89:312–323

    Article  CAS  Google Scholar 

  16. Panja D, Kenney JW, D’Andrea L, Zalfa F, Vedeler A, Wibrand K et al (2014) Two-stage translational control of dentate gyrus LTP consolidation is mediated by sustained BDNF-TrkB signaling to MNK. Cell Rep 9:1430–1445

    Article  CAS  Google Scholar 

  17. Schratt GM, Nigh EA, Chen WG, Hu L, Greenberg ME (2004) BDNF regulates the translation of a select group of mRNAs by a mammalian target of rapamycin-phosphatidylinositol 3-kinase-dependent pathway during neuronal development. J Neurosci 24:7366–7377

    Article  CAS  Google Scholar 

  18. Santos AR, Comprido D, Duarte CB (2010) Regulation of local translation at the synapse by BDNF. Prog Neurobiol 92:505–516

    Article  CAS  Google Scholar 

  19. Gingras AC, Raught B, Sonenberg N (1999) eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. Annu Rev Biochem 68:913–963

    Article  CAS  Google Scholar 

  20. Faye MD, Graber TE, Holcik M (2014) Assessment of selective mRNA translation in mammalian cells by polysome profiling. J Vis Exp e52295

    Google Scholar 

  21. Bradshaw KD, Emptage NJ, Bliss TV (2003) A role for dendritic protein synthesis in hippocampal late LTP. Eur J Neurosci 18:3150–3152

    Article  CAS  Google Scholar 

  22. Bramham CR, Jensen KB, Proud CG (2016) Tuning specific translation in cancer metastasis and synaptic memory: control at the MNK-eIF4E Axis. Trends Biochem Sci 41:847–858

    Article  CAS  Google Scholar 

  23. Mader S, Lee H, Pause A, Sonenberg N (1995) The translation initiation factor eIF-4E binds to a common motif shared by the translation factor eIF-4 gamma and the translational repressors 4E-binding proteins. Mol Cell Biol 15:4990–4997

    Article  CAS  Google Scholar 

  24. Nathans D (1964) Puromycin inhibition of protein synthesis: incorporation of Puromycin into peptide chains. Proc Natl Acad Sci U S A 51:585–592

    Article  CAS  Google Scholar 

  25. Nathans D (1964) Inhibition of protein synthesis by puromycin. Fed Proc 23:984–989

    CAS  PubMed  Google Scholar 

  26. Schmidt EK, Clavarino G, Ceppi M, Pierre P (2009) SUnSET, a nonradioactive method to monitor protein synthesis. Nat Methods 6:275–277

    Article  CAS  Google Scholar 

  27. Alvarez-Castelao B, Schuman EM (2015) The regulation of synaptic protein turnover. J Biol Chem 290:28623–28630

    Article  CAS  Google Scholar 

  28. Gal-Ben-Ari S, Kenney JW, Ounalla-Saad H, Taha E, David O, Levitan D et al (2012) Consolidation and translation regulation. Learn Mem 19:410–422

    Article  CAS  Google Scholar 

  29. Ingolia NT (2014) Ribosome profiling: new views of translation, from single codons to genome scale. Nat Rev Genet 15:205–213

    Article  CAS  Google Scholar 

  30. Niranjanakumari S, Lasda E, Brazas R, Garcia-Blanco MA (2002) Reversible cross-linking combined with immunoprecipitation to study RNA-protein interactions in vivo. Methods 26:182–190

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Institute of Mental Health, Bethesda, MD, USA

    Debabrata Panja

  2. Department of Biomedicine and KG Jebsen Center for Research on Neuropsychiatric Disorders, University of Bergen, Bergen, Norway

    Clive R. Bramham

Authors
  1. Debabrata Panja
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Clive R. Bramham
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Debabrata Panja .

Editor information

Editors and Affiliations

  1. Center for Neuroscience and Cell Biology and Department of Life Sciences, University of Coimbra, Coimbra, Portugal

    Carlos B. Duarte

  2. BRAIN Center for Neuroscience, Department of Life Sciences, University of Trieste, Trieste, Italy

    Enrico Tongiorgi

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Panja, D., Bramham, C.R. (2017). BDNF Function in Long-Term Synaptic Plasticity in the Dentate Gyrus In Vivo: Methods for Local Drug Delivery and Biochemical Analysis of Translation. In: Duarte, C., Tongiorgi, E. (eds) Brain-Derived Neurotrophic Factor (BDNF). Neuromethods, vol 143. Humana, New York, NY. https://doi.org/10.1007/7657_2017_7

Download citation

  • DOI: https://doi.org/10.1007/7657_2017_7

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-8969-0

  • Online ISBN: 978-1-4939-8970-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation