Recordings of cultured neurons and synaptic activity using patch-clamp chips

Area of Science:

• Neuroscience
• Biotechnology
• Pharmacology

Background:

• Planar patch-clamp chip technology advances drug efficacy and safety assessments.
• Current limitations restrict recordings to isolated cells, hindering synaptic transmission studies.
• Previous work demonstrated planar patch-clamp chips for recording from cultured neurons.

Purpose of the Study:

• To establish and record from a reconstructed synaptic circuit using planar patch-clamp chips.
• To investigate synaptic phenomena in synaptically connected neurons cultured on-chip.
• To demonstrate the utility of this technology for studying synaptic communication and pharmacology.

Main Methods:

• Development of single- and dual-recording site planar patch-clamp chips.
• Reconstruction of a pre-synaptic and post-synaptic neuronal circuit (visceral dorsal 4 and left pedal dorsal 1 neurons) from Lymnaea stagnalis.
• On-chip cytoplasmic perfusion using subterranean microfluidics for individual neuron manipulation.
• Pharmacological characterization of the identified synapse.

Main Results:

• First successful planar patch-clamp chip recordings of synaptic phenomena from paired cultured neurons.
• Demonstration of cholinergic neurotransmission at the reconstructed synapse.
• Simultaneous dual-site recordings from paired neurons.
• Successful dedicated cytoplasmic perfusion of individual neurons.

Conclusions:

• Planar patch-clamp chip technology is now applicable to studying synaptic communication.
• This technology provides a novel platform for investigating synaptic transmission and pharmacology in complex neural circuits.
• The developed on-chip system enables detailed analysis of synaptic function and drug interactions.