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BDNF-Induced Intracellular Signaling

  • João R. GomesEmail author
  • Andrea Lobo
  • Carlos B. Duarte
  • Mário Grãos
Protocol
Part of the Neuromethods book series (NM, volume 143)

Abstract

The neurotrophin BDNF plays important roles in neuronal survival, growth, and differentiation during development. Furthermore, it has been shown to mediate long-term changes in the synaptic activity in the hippocampus and in other brain regions, which are thought to underlie learning and memory formation. Cultured hippocampal neurons express TrkB receptors and, therefore, constitute a valuable experimental model to study in vitro BDNF-induced intracellular signaling pathways. In this chapter, we describe (1) the methodology used to prepare cultured hippocampal neurons from mice (wild-type-WT or transgenic animals) and rats and (2) three different approaches to investigate BDNF signaling: Western blot and Bio-Plex for overall signaling activity and immunocytochemistry to analyze where signaling activity takes place in neurons. The Bio-Plex approach allows the simultaneous characterization of different pathways using small sample volumes and within a short period of time.

Keywords

Akt Bio-Plex Cultured hippocampal neurons ERK Immunocytochemistry Phospholipase C Western blot 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • João R. Gomes
    • 1
    • 2
    Email author
  • Andrea Lobo
    • 1
    • 3
  • Carlos B. Duarte
    • 4
    • 5
  • Mário Grãos
    • 4
  1. 1.Instituto de Investigação e Inovação em Saúde (I3S)University of PortoPortoPortugal
  2. 2.Molecular Neurobiology, IBMC-Institute for Molecular and Cell BiologyUniversity of PortoPortoPortugal
  3. 3.Addiction Biology, IBMC-Institute for Molecular and Cell BiologyUniversity of PortoPortoPortugal
  4. 4.CNC-Center for Neuroscience and Cell BiologyUniversity of CoimbraCoimbraPortugal
  5. 5.Department of Life SciencesUniversity of CoimbraCoimbraPortugal

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