Microfluidic Local Perfusion Chambers for the Visualization and Manipulation of Synapses
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 (μLP) 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.
Additional Information
© 2010 Elsevier. Accepted 17 February 2010. Published: April 14, 2010. Available online 14 April 2010. The authors gratefully acknowledge Lin Chen for the hippocampal neuron cultures, Michael Roukes for use of his clean room, Julie Cho for help with preliminary experiments, and Jennifer Hodas for reviewing the manuscript. A.M.T. received a Johnson & Johnson fellowship. We acknowledge support from the Weston Havens Foundation, NIH, and Howard Hughes Medical Institute. A.M.T. is a cofounder of Xona Microfluidics, LLC.Additional details
- Eprint ID
- 18521
- DOI
- 10.1016/j.neuron.2010.03.022
- Resolver ID
- CaltechAUTHORS:20100602-114028118
- Johnson & Johnson fellowship
- Weston Havens Foundation
- NIH
- Howard Hughes Medical Institute
- Created
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2010-06-02Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field