title: 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 (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.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Taylor
authors.forename: Daniela C
authors.initials: DC
authors.lastname: Dieterich
authors.forename: Hiroshi T
authors.initials: HT
authors.lastname: Ito
authors.forename: Sally A
authors.initials: SA
authors.lastname: Kim
authors.forename: Erin M
authors.initials: EM
authors.lastname: Schuman
mesh_terms: 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*
references.citation: Nature. 2009 Mar 19;458(7236):299-304
references.pmid: 19295602
references.citation: Biomaterials. 2005 May;26(15):2549-57
references.pmid: 15585257
references.citation: Annu Rev Biomed Eng. 2001;3:335-73
references.pmid: 11447067
references.citation: Science. 2000 Apr 7;288(5463):113-6
references.pmid: 10753110
references.citation: Neuron. 2008 Aug 14;59(3):475-85
references.pmid: 18701072
references.citation: J Neurosci Methods. 2000 Dec 15;104(1):65-75
references.pmid: 11163412
references.citation: Neuron. 2008 Jul 10;59(1):84-97
references.pmid: 18614031
references.citation: Neuron. 2008 Jul 10;59(1):70-83
references.pmid: 18614030
references.citation: Neuron. 1998 Oct;21(4):741-51
references.pmid: 9808461
references.citation: Nature. 2007 Apr 5;446(7136):633-9
references.pmid: 17410168
references.citation: Neuron. 2001 May;30(2):489-502
references.pmid: 11395009
references.citation: Methods Mol Biol. 2006;321:167-77
references.pmid: 16508072
references.citation: Science. 1972 Jan 28;175(4020):451-4
references.pmid: 17731371
references.citation: Science. 2008 Jul 4;321(5885):136-40
references.pmid: 18556515
references.citation: Langmuir. 2003 Mar 4;19(5):1551-1556
references.pmid: 20725530
references.citation: J Neurosci Methods. 2004 Apr 30;134(2):191-8
references.pmid: 15003385
references.citation: Cell. 2006 May 19;125(4):785-99
references.pmid: 16713568
references.citation: Conf Proc IEEE Eng Med Biol Soc. 2004;2004:2542-5
references.pmid: 17270791
references.citation: Nat Biotechnol. 2002 Aug;20(8):826-30
references.pmid: 12091913
references.citation: Nat Methods. 2007 Sep;4(9):727-31
references.pmid: 17704783
references.citation: Electrophoresis. 2000 Jan;21(1):27-40
references.pmid: 10634468
references.citation: J Neurosci. 1998 Jan 1;18(1):26-35
references.pmid: 9412483
references.citation: Nat Neurosci. 2002 Apr;5(4):348-55
references.pmid: 11889468
references.citation: J Neurosci Methods. 1998 Aug 1;82(2):167-73
references.pmid: 9700689
references.citation: Cell. 1994 Oct 7;79(1):49-58
references.pmid: 7923376
references.citation: J Neurosci. 2009 Apr 15;29(15):4697-707
references.pmid: 19369540
references.citation: Nature. 2006 Jul 27;442(7101):403-11
references.pmid: 16871208
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):12542-7
references.pmid: 12237407
references.citation: Nat Methods. 2005 Aug;2(8):599-605
references.pmid: 16094385
references.citation: Neuron. 2007 Aug 16;55(4):648-61
references.pmid: 17698016
references.citation: Nature. 1997 Jul 17;388(6639):275-9
references.pmid: 9230436
references.citation: Nat Neurosci. 2001 Feb;4(2):151-8
references.pmid: 11175875
references.citation: Conf Proc IEEE Eng Med Biol Soc. 2006;2006:4322-5
references.pmid: 17946237