title: Targeted electrophysiological analysis of viscerofugal neurons in the myenteric plexus of guinea-pig colon.
abstract: Enteric viscerofugal neurons are mechanosensory interneurons that form the afferent limb of intestino-intestinal reflexes involving prevertebral sympathetic neurons. Fast synaptic inputs to viscerofugal neurons arise from other enteric neurons, but their sources are unknown. We aimed to describe the origins of synaptic inputs to viscerofugal neurons by mapping the locations of their cell bodies within the myenteric plexus. Viscerofugal neuron somata were retrogradely traced with 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) from colonic nerve trunks and impaled with microelectrodes, in longitudinal muscle/myenteric plexus preparations of the guinea-pig distal colon (39 impalements, n=14). Thirty-eight viscerofugal neurons were uni-axonal and had the electrophysiological characteristics of myenteric S-neurons; one neuron was multipolar with AH-neuron electrophysiological characteristics. Depolarizing current pulses evoked either single- or multiple action potentials in viscerofugal neurons (range 1-25 spikes, 500 ms, 100-900 pA, 21 cells). Electrical stimulation of internodal strands circumferential to viscerofugal neurons evoked fast excitatory postsynaptic potentials (EPSPs) in 19/24 cells. Focal pressure-ejection of the nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP, 10 μm) directly onto viscerofugal nerve cell bodies evoked large depolarizations and action potentials (23 ± 10 mV, latency 350 ± 230 ms, 21/22 cells). DMPP was then focally applied to multiple sites, up to 3mm from the recorded viscerofugal neuron, to activate other myenteric S-neurons. In a few sites in myenteric ganglia, DMPP evoked repeatable fast EPSPs in viscerofugal neurons (latency 300 ± 316 ms, 38/394 sites, 10 cells). The cellular sources of synaptic inputs to viscerofugal neurons were located both orally and aborally (19 oral, 19 aboral), but the amplitude of oral inputs was consistently greater than aboral inputs (13.1 ± 4.3 mV vs. 10.1 ± 4.8 mV, respectively, p<0.05, paired t-test, n=6). Most impaled viscerofugal neurons were nitric oxide synthase (NOS) immunoreactive (20/27 cells tested). Thus, the synaptic connections onto viscerofugal neurons within the myenteric plexus suggest that multiple enteric neural pathways feed into intestino-intestinal reflexes, involving sympathetic prevertebral ganglia.
authors.affiliation: Human Physiology, Flinders Medical Science and Technology, School of Medicine and Centre for Neuroscience, Flinders University, Bedford Park, South Australia 5042, Australia.
authors.forename: T J
authors.initials: TJ
authors.lastname: Hibberd
authors.affiliation: Human Physiology, Flinders Medical Science and Technology, School of Medicine and Centre for Neuroscience, Flinders University, Bedford Park, South Australia 5042, Australia.
authors.forename: N J
authors.initials: NJ
authors.lastname: Spencer
authors.affiliation: Human Physiology, Flinders Medical Science and Technology, School of Medicine and Centre for Neuroscience, Flinders University, Bedford Park, South Australia 5042, Australia.
authors.forename: V P
authors.initials: VP
authors.lastname: Zagorodnyuk
authors.affiliation: Human Physiology, Flinders Medical Science and Technology, School of Medicine and Centre for Neuroscience, Flinders University, Bedford Park, South Australia 5042, Australia.
authors.forename: B N
authors.initials: BN
authors.lastname: Chen
authors.affiliation: Human Physiology, Flinders Medical Science and Technology, School of Medicine and Centre for Neuroscience, Flinders University, Bedford Park, South Australia 5042, Australia. Electronic address: simon.brookes@flinders.edu.au.
authors.forename: S J H
authors.initials: SJ
authors.lastname: Brookes
mesh_terms: D000818:Animals; D003106:Colon / Q000294:innervation*; D004558:Electric Stimulation; D055724:Electrophysiological Phenomena; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D005728:Ganglia, Sympathetic / Q000502:physiology; D006168:Guinea Pigs; D007150:Immunohistochemistry; D009197:Myenteric Plexus / Q000502:physiology*; D009434:Neural Pathways / Q000502:physiology; D009474:Neurons / Q000502:physiology*
keywords: cholinergic; enteric nervous system; sensory neurons; sympathetic nervous system; synaptic potentials