Generation of Functional Inhibitory Synapses Incorporating Defined Combinations of GABA(A) or Glycine Receptor Subunits

Frontiers in Molecular Neuroscience
Christine L DixonJoseph W Lynch

Abstract

Fast inhibitory neurotransmission in the brain is mediated by wide range of GABAA receptor (GABAAR) and glycine receptor (GlyR) isoforms, each with different physiological and pharmacological properties. Because multiple isoforms are expressed simultaneously in most neurons, it is difficult to define the properties of individual isoforms under synaptic stimulation conditions in vivo. Although recombinant expression systems permit the expression of individual isoforms in isolation, they require exogenous agonist application which cannot mimic the dynamic neurotransmitter profile characteristic of native synapses. We describe a neuron-HEK293 cell co-culture technique for generating inhibitory synapses incorporating defined combinations of GABAAR or GlyR subunits. Primary neuronal cultures, prepared from embryonic rat cerebral cortex or spinal cord, are used to provide presynaptic GABAergic and glycinergic terminals, respectively. When the cultures are mature, HEK293 cells expressing the subunits of interest plus neuroligin 2A are plated onto the neurons, which rapidly form synapses onto HEK293 cells. Patch clamp electrophysiology is then used to analyze the physiological and pharmacological properties of the inhibitory postsynapt...Continue Reading

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Citations

Oct 8, 2016·Frontiers in Molecular Neuroscience·Yan ZhangJoseph W Lynch
Aug 2, 2017·Epilepsia·Mayara Vendramin PasquettiMaria Elisa Calcagnotto
Jun 8, 2017·Frontiers in Molecular Neuroscience·Yan ZhangAngelo Keramidas
May 31, 2018·Proceedings of the National Academy of Sciences of the United States of America·Géza BereckiSteven Petrou
Oct 27, 2020·Epilepsia·Eleonora LugaràMatthew C Walker
Dec 18, 2020·Frontiers in Molecular Neuroscience·Parnayan SyedJoseph W Lynch
Mar 24, 2021·The Journal of General Physiology·Chung-Wei ChiangMeyer B Jackson

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