A Microfluidic Culture Platform for Neurotrophin Signaling Studies
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.
KeywordsAxons BDNF Compartmentalized cultures Dendrites Dorsal root ganglia (DRG) Microfluidic chambers Neurotrophin NGF Retrograde labeling Retrograde signaling
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