Microfluidic local perfusion chambers for the visualization and manipulation of synapses.
The polarized nature of neurons and the size and density of synapses complicates the manipulation and visualization of cell biological processes that control synaptic function. Here we developed a microfluidic local perfusion (microLP) chamber to access and manipulate synaptic regions and presynaptic and postsynaptic compartments in vitro. This chamber directs the formation of synapses in >100 parallel rows connecting separate neuron populations. A perfusion channel transects the parallel rows, allowing access with high spatial and temporal resolution to synaptic regions. We used this chamber to investigate synapse-to-nucleus signaling. Using the calcium indicator dye Fluo-4 NW, we measured changes in calcium at dendrites and somata, following local perfusion of glutamate. Exploiting the high temporal resolution of the chamber, we exposed synapses to "spaced" or "massed" application of glutamate and then examined levels of pCREB in somata. Lastly, we applied the metabotropic receptor agonist DHPG to dendrites and observed increases in Arc transcription and Arc transcript localization.
Taylor
Anne M
AM
Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
Dieterich
Daniela C
DC
Ito
Hiroshi T
HT
Kim
Sally A
SA
Schuman
Erin M
EM
D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation; D003712:Dendrites / Q000502:physiology*; D004867:Equipment Design; D005110:Extracellular Space; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D009446:Neurobiology / Q000295:instrumentation* / Q000379:methods; D010477:Perfusion / Q000295:instrumentation; D017729:Presynaptic Terminals / Q000502:physiology; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D055366:Synaptic Potentials / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology*
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