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Achieving synaptically relevant pulses of neurotransmitter using PDMS microfluidics
E J Botzolakis1, A Maheshwari, H J Feng
1Graduate Program in Neuroscience, Vanderbilt University, 465 21st Avenue South, MRBIII, Suite 6140, Nashville, TN 37232, United States. manuel.botzolakis@vanderbilt.edu <manuel.botzolakis@vanderbilt.edu>
Journal of Neuroscience Methods
|November 18, 2008
Summary
Researchers developed a novel microfluidic system for precise, rapid neurotransmitter application. This method enables studying ligand-gated ion channels (LGICs) under conditions that mimic synaptic activity, improving disease research.
Area of Science:
- Neuroscience
- Molecular Biology
- Biophysics
Background:
- Fast synaptic transmission relies on ligand-gated ion channels (LGICs) activated by transient neurotransmitter release.
- Studying LGICs in vitro under physiologically relevant, synaptic timescales remains challenging.
- Disruptions in synaptic transmission are linked to neurological and psychiatric disorders.
Purpose of the Study:
- To introduce a novel microfluidic solution-switching system for precise temporal control of neurotransmitter application.
- To enable the study of LGICs under synaptically relevant conditions with enhanced experimental throughput and reproducibility.
- To investigate the functional properties of GABA(A) receptors using ultra-brief neurotransmitter pulses.
Main Methods:
- Development of a microfluidic device for rapid solution switching, enabling precise temporal control over neurotransmitter application.
- Application of ultra-brief (approximately 400 microseconds) GABA pulses to recombinant GABA(A) receptors.
- Comparison of receptor currents evoked by brief synaptic-like pulses versus conventional longer pulses.
Main Results:
- The microfluidic system achieved precise temporal control over neurotransmitter application, mimicking synaptic transients.
- GABA pulses applied via the system to GABA(A) receptors generated currents resembling hippocampal inhibitory post-synaptic currents (IPSCs).
- Currents evoked by brief pulses differed significantly from those evoked by longer, conventional pulses, highlighting the importance of timescale.
Conclusions:
- The novel microfluidic approach provides a practical and effective method for studying LGICs under synaptically relevant conditions.
- This methodology facilitates the evaluation of disease-causing mutations and allosteric modulators in vitro.
- The findings underscore the critical importance of assessing LGIC function on a synaptic timescale for accurate physiological relevance.
