Microfluidic Local Perfusion Chambers for the Visualization and Manipulation of Synapses

Overview

Journal Neuron

Publisher Cell Press

Specialty Neurology

Date 2010 Apr 20

PMID 20399729

Citations 130

Authors

Anne M Taylor

Daniela C Dieterich

Hiroshi T Ito

Sally A Kim

Erin M Schuman

Affiliations

Soon will be listed here.

Abstract

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.

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