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.

[1]  S. Quake,et al.  Monolithic microfabricated valves and pumps by multilayer soft lithography. , 2000, Science.

[2]  K. Jensen,et al.  Cells on chips , 2006, Nature.

[3]  Y.-C. Tai,et al.  Development of biocompatible parylene neurocages , 2004, The 26th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[4]  Satoshi Kida,et al.  CREB required for the stability of new and reactivated fear memories , 2002, Nature Neuroscience.

[5]  Mu-ming Poo,et al.  cAMP-induced switching in turning direction of nerve growth cones , 1997, Nature.

[6]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[7]  Richard L. Huganir,et al.  Elongation Factor 2 and Fragile X Mental Retardation Protein Control the Dynamic Translation of Arc/Arg3.1 Essential for mGluR-LTD , 2008, Neuron.

[8]  Karl Deisseroth,et al.  Spaced stimuli stabilize MAPK pathway activation and its effects on dendritic morphology , 2001, Nature Neuroscience.

[9]  Eric R. Kandel,et al.  Long-Term Habituation of a Defensive Withdrawal Reflex in Aplysia , 1972, Science.

[10]  Carl W. Cotman,et al.  Microlithographic determination of axonal/dendritic polarity in cultured hippocampal neurons , 1998, Journal of Neuroscience Methods.

[11]  Feng Zhang,et al.  Multimodal fast optical interrogation of neural circuitry , 2007, Nature.

[12]  P. Worley,et al.  Differential Intracellular Sorting of Immediate Early Gene mRNAs Depends on Signals in the mRNA Sequence , 1998, The Journal of Neuroscience.

[13]  Yu-Chong Tai,et al.  Parylene Neurocages for Electrical Stimulation on Silicon and Glass Substrates , 2006, 2006 International Conference of the IEEE Engineering in Medicine and Biology Society.

[14]  C. Cotman,et al.  A microfluidic culture platform for CNS axonal injury, regeneration and transport , 2005, Nature Methods.

[15]  Brad E. Pfeiffer,et al.  Rapid Translation of Arc/Arg3.1 Selectively Mediates mGluR-Dependent LTD through Persistent Increases in AMPAR Endocytosis Rate , 2008, Neuron.

[16]  Noo Li Jeon,et al.  Microfluidic chambers for cell migration and neuroscience research. , 2006, Methods in molecular biology.

[17]  Oswald Steward,et al.  Synaptic Activation Causes the mRNA for the IEG Arc to Localize Selectively near Activated Postsynaptic Sites on Dendrites , 1998, Neuron.

[18]  Erin M. Schuman,et al.  Dynamic Visualization of Local Protein Synthesis in Hippocampal Neurons , 2001, Neuron.

[19]  M Krause,et al.  Ordered networks of rat hippocampal neurons attached to silicon oxide surfaces , 2000, Journal of Neuroscience Methods.

[20]  Karel Svoboda,et al.  The Spread of Ras Activity Triggered by Activation of a Single Dendritic Spine , 2008, Science.

[21]  W. Quinn,et al.  Induction of a dominant negative CREB transgene specifically blocks long-term memory in Drosophila , 1994, Cell.

[22]  Andreas Offenhäusser,et al.  Impact of micropatterned surfaces on neuronal polarity , 2004, Journal of Neuroscience Methods.

[23]  G. Whitesides,et al.  Fabrication of microfluidic systems in poly(dimethylsiloxane) , 2000, Electrophoresis.

[24]  G. Whitesides,et al.  Gradients of substrate-bound laminin orient axonal specification of neurons , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  E. Schuman,et al.  Postsynaptic Decoding of Neural Activity: eEF2 as a Biochemical Sensor Coupling Miniature Synaptic Transmission to Local Protein Synthesis , 2007, Neuron.

[26]  M. Poo,et al.  Erratum: CAMP-induced switching in turning direction of nerve growth cones (Nature (1997) 388 (275-279)) , 1997 .

[27]  Andreas Offenhäusser,et al.  Synaptic plasticity in micropatterned neuronal networks. , 2005, Biomaterials.

[28]  Carl W. Cotman,et al.  Axonal mRNA in Uninjured and Regenerating Cortical Mammalian Axons , 2009, The Journal of Neuroscience.

[29]  G. Whitesides,et al.  Neutrophil chemotaxis in linear and complex gradients of interleukin-8 formed in a microfabricated device , 2002, Nature Biotechnology.

[30]  Seok-Jin R. Lee,et al.  Activation of CaMKII in single dendritic spines during long-term potentiation , 2009, Nature.

[31]  T. Branco,et al.  Local Dendritic Activity Sets Release Probability at Hippocampal Synapses , 2008, Neuron.

[32]  Cori Bargmann,et al.  Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans , 2007, Nature Methods.

[33]  E. Schuman,et al.  Miniature Neurotransmission Stabilizes Synaptic Function via Tonic Suppression of Local Dendritic Protein Synthesis , 2006, Cell.

[34]  Noo Li Jeon,et al.  Microfluidic Multicompartment Device for Neuroscience Research. , 2003, Langmuir : the ACS journal of surfaces and colloids.