Protocol to correlate electron microscopy with electrophysiology in single-cell autaptic microcultures.

Martínez San Segundo P., Pérez González APL., Velasco CD., Llobet A.

Single-cell microcultures (SCMs) form a monosynaptic circuit that allows stimulation and recording of postsynaptic responses using a single electrode. Here, we present a protocol to establish autaptic cultures from rat superior cervical ganglion neurons. We describe the steps for preparing SCMs, recording synaptic currents, and identifying and processing the recorded neurons for electron microscopy. We then detail procedures for visualizing synapses. This protocol is illustrated by correlating evoked and spontaneous neurotransmitter release with the ultrastructural features of synapses recorded. For complete details on the use and execution of this protocol, please refer to Velasco et al.1.

DOI

10.1016/j.xpro.2024.103003

Type

Journal article

Publication Date

2024-06-21T00:00:00+00:00

Volume

5

Keywords

Cell Biology, Cell culture, Neuroscience, Single Cell, Animals, Rats, Neurons, Microscopy, Electron, Synapses, Electrophysiology, Cell Culture Techniques, Superior Cervical Ganglion, Cells, Cultured, Electrophysiological Phenomena, Single-Cell Analysis

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