Achieving synaptically relevant pulses of neurotransmitter using PDMS microfluidics.

Botzolakis EJ, Maheshwari A, Feng HJ, Lagrange AH, Shaver JH, Kassebaum NJ, Venkataraman R, Baudenbacher F, Macdonald RL
J Neurosci Methods. 2009 177 (2): 294-302

PMID: 19013195 · PMCID: PMC3786708 · DOI:10.1016/j.jneumeth.2008.10.014

Fast synaptic transmission is mediated by post-synaptic ligand-gated ion channels (LGICs) transiently activated by neurotransmitter released from pre-synaptic vesicles. Although disruption of synaptic transmission has been implicated in numerous neurological and psychiatric disorders, effective and practical methods for studying LGICs in vitro under synaptically relevant conditions are unavailable. Here, we describe a novel microfluidic approach to solution switching that allows for precise temporal control over the neurotransmitter transient while substantially increasing experimental throughput, flexibility, reproducibility, and cost-effectiveness. When this system was used to apply ultra-brief ( approximately 400micros) GABA pulses to recombinant GABA(A) receptors, members of the cys-loop family of LGICs, the resulting currents resembled hippocampal inhibitory post-synaptic currents (IPSCs) and differed from currents evoked by longer, conventional pulses, illustrating the importance of evaluating LGICs on a synaptic timescale. This methodology should therefore allow the effects of disease-causing mutations and allosteric modulators to be evaluated in vitro under physiologically relevant conditions.

MeSH Terms (17)

Allosteric Regulation Cell Line Drug Delivery Systems Electronics, Medical Electrophysiology Humans Inhibitory Postsynaptic Potentials Microfluidic Analytical Techniques Neural Inhibition Neurochemistry Neurotransmitter Agents Patch-Clamp Techniques Presynaptic Terminals Receptors, GABA-A Recombinant Proteins Synaptic Transmission Time Factors

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