pp 1-17 | Cite as

A Microfluidic Culture Platform for Neurotrophin Signaling Studies

  • Rui O. Costa
  • Tânia Perestrelo
  • Diogo Tomé
  • Ramiro D. Almeida
Protocol
Part of the Neuromethods book series

Abstract

Brain-derived neurotrophic factor (BDNF) has a crucial role in neuronal development, maintenance, and function. Some of the effects induced by the neurotrophins occur at neuronal sub-domains, which are located at significant distances from the cell body. The study of the mechanisms involved in the axonal responses to BDNF, as well as to other pro- and mature neurotrophins, requires a reliable system, to physically isolate these sub-domains from the soma in order to better evaluate its contribution to the neuronal response. Here we describe how to culture and maintain dorsal root ganglia (DRG) neurons in microfluidic chambers and demonstrate the physical and fluidic isolation of pure axonal populations (distal axons) in this platform. In addition, we describe how to independently manipulate distal axons or the somal compartment and how to retrogradely label the targeted neurons. In this chapter we describe how this powerful tool can be used to study the effect of neurotrophins in spatially isolated axons.

Keywords

Axons BDNF Compartmentalized cultures Dendrites Dorsal root ganglia (DRG) Microfluidic chambers Neurotrophin NGF Retrograde labeling Retrograde signaling 

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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  • Rui O. Costa
    • 1
    • 2
  • Tânia Perestrelo
    • 1
  • Diogo Tomé
    • 1
  • Ramiro D. Almeida
    • 1
    • 2
    • 3
  1. 1.CNC—Center for Neuroscience and Cell BiologyUniversity of CoimbraCoimbraPortugal
  2. 2.Institute for Interdisciplinary ResearchUniversity of CoimbraCoimbraPortugal
  3. 3.Department of Medical Sciences and Institute of Biomedicine—iBiMEDUniversity of AveiroAveiroPortugal

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