Isolation and Electrophysiology of Murine Sympathetic Postganglionic Neurons in the Thoracic Paravertebral Ganglia

• Type of Material: Article
• Authors: Mallika Halder, Emory University; Michael Lee McKinnon, Emory University; Yaqing Li, Emory University; Peter Wenner, Emory University; Shawn Hochman, Emory University
• Language: English
• Date: 2021-10-20
• Publisher: Bio-protocol
• Publication Version: Final Published Version
• Copyright Statement: © 2021 The Authors; exclusive licensee Bio-protocol LLC.
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.21769/BioProtoc.4189
• Title of Journal or Parent Work: Bio-protocol
• Volume: 11
• Issue: 20
• Start Page: e4189
• End Page: e4189
• Grant/Funding Information: This work was supported by the National Institutes of Health Grant 5R01NS102871 and the Department of Defense Grant SCI-30225.
• Keywords: Sympathetic chain; Postganglionic neuron; Preganglionic neuron; Extracellular electrode; Paravertebral ganglia; Optogenetic; Mouse
• Research Categories: Health Sciences, Medicine and Surgery

Abstract

[Abstract] The thoracic paravertebral sympathetic chain postganglionic neurons (tSPNs) represent the predominant sympathetic control of vascular function in the trunk and upper extremities. tSPNs cluster to form ganglia linked by an interganglionic nerve and receive multisegmental convergent and divergent synaptic input from cholinergic sympathetic preganglionic neurons of the spinal cord (Blackman and Purves, 1969; Lichtman et al., 1980). Studies in the past have focused on cervical and lumbar chain ganglia in multiple species, but few have examined the thoracic chain ganglia, whose location and diminutive size make them less conducive to experimentation. Seminal studies on the integrative properties of preganglionic axonal projections onto tSPNs were performed in guinea pig (Blackman and Purves, 1969; Lichtman et al., 1980), but as mice have become the accepted mammalian genetic model organism, there is need to reproduce and expand on these studies in this smaller model. We describe an ex vivo approach that enables electrophysiological, calcium imaging, and optogenetic assessment of convergence, divergence, and studies on pre- to postganglionic synaptic transmission, as well as whole-cell recordings from individual tSPNs. Preganglionic axonal connections from intact ventral roots and interganglionic nerves across multiple segments can be stimulated to evoke compound action potential responses in individual thoracic ganglia as recorded with suction electrodes. Chemical block of synaptic transmission differentiates spiking of preganglionic axons from synaptically-recruited tSPNs. Further dissection, including removal of the sympathetic chain, enables whole-cell patch clamp recordings from individual tSPNs for characterization of cellular and synaptic properties.