Mechanisms of orexin-induced depolarizations in rat dorsal motor nucleus of vagus neurones in vitro

Ling Ling Hwang, Chiung Tong Chen, Nae J. Dun

Research output: Contribution to journalArticle

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Abstract

1. Whole-cell patch-clamp recordings were made from neurones of the dorsal motor nucleus of the vagus (DMNV), including Fluoro-gold-labelled parasympathetic preganglionic neurones (PPNs), in slices of the rat medulla. In the latter case, rats had received an I.P. injection of Fluoro-gold solution (10 μg) 2-3 days earlier. 2. Superfusion of orexin A or B (10-300 nM) caused a slow depolarization in approximately 30% of the DMNV neurones, including PPNs. Orexin-induced depolarizations, which persisted in TTX (0.5 μM)-containing Krebs solution, were reduced by 70% in a low-Na+ (26 mM) Krebs solution, indicating the involvement of Na+ ions. A significant change in orexin-induced depolarizations was not obtained in either a high-K+ (7 mM) or Cd2+ (100 μM) Krebs solution. 3. Inclusion of the hydrolysis-resistant guanine nucleotide GDP-β-S in the patch solution significantly reduced the orexin A- or B-induced depolarizations. 4. Under whole-cell voltage-clamp conditions, the orexin-induced inward current declined with hyperpolarization, but did not reverse polarity in the potential range between - 120 and 0 mV. In low-Na+ solution, the orexin-induced current was reduced and the I- V curve reversed polarity at about - 105 mV; the response was further reduced and the reversal potential shifted to -90 mV in a low-Na+, high-K+ Krebs solution. 5. It is concluded that the peptides orexin A and B, acting on orexin receptors, which are GTP-binding-protein coupled, are excitatory to DMNV neurones. In addition, more than one conductance, which may include a non-selective cation conductance and a K+ conductance, appears to be involved in the orexin-induced depolarization.

Original languageEnglish
Pages (from-to)511-520
Number of pages10
JournalJournal of Physiology
Volume537
Issue number2
DOIs
Publication statusPublished - Dec 1 2001

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Neurons
Orexin Receptors
Guanine Nucleotides
Motor Neurons
GTP-Binding Proteins
Orexins
In Vitro Techniques
Cations
Hydrolysis
Ions
Injections
Krebs-Ringer solution
methanesulfonate salt 2-hydroxy-4,4'-diamidinostilbene

ASJC Scopus subject areas

  • Physiology

Cite this

Mechanisms of orexin-induced depolarizations in rat dorsal motor nucleus of vagus neurones in vitro. / Hwang, Ling Ling; Chen, Chiung Tong; Dun, Nae J.

In: Journal of Physiology, Vol. 537, No. 2, 01.12.2001, p. 511-520.

Research output: Contribution to journalArticle

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AB - 1. Whole-cell patch-clamp recordings were made from neurones of the dorsal motor nucleus of the vagus (DMNV), including Fluoro-gold-labelled parasympathetic preganglionic neurones (PPNs), in slices of the rat medulla. In the latter case, rats had received an I.P. injection of Fluoro-gold solution (10 μg) 2-3 days earlier. 2. Superfusion of orexin A or B (10-300 nM) caused a slow depolarization in approximately 30% of the DMNV neurones, including PPNs. Orexin-induced depolarizations, which persisted in TTX (0.5 μM)-containing Krebs solution, were reduced by 70% in a low-Na+ (26 mM) Krebs solution, indicating the involvement of Na+ ions. A significant change in orexin-induced depolarizations was not obtained in either a high-K+ (7 mM) or Cd2+ (100 μM) Krebs solution. 3. Inclusion of the hydrolysis-resistant guanine nucleotide GDP-β-S in the patch solution significantly reduced the orexin A- or B-induced depolarizations. 4. Under whole-cell voltage-clamp conditions, the orexin-induced inward current declined with hyperpolarization, but did not reverse polarity in the potential range between - 120 and 0 mV. In low-Na+ solution, the orexin-induced current was reduced and the I- V curve reversed polarity at about - 105 mV; the response was further reduced and the reversal potential shifted to -90 mV in a low-Na+, high-K+ Krebs solution. 5. It is concluded that the peptides orexin A and B, acting on orexin receptors, which are GTP-binding-protein coupled, are excitatory to DMNV neurones. In addition, more than one conductance, which may include a non-selective cation conductance and a K+ conductance, appears to be involved in the orexin-induced depolarization.

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