Strategies for cellular identification in nucleus tractus solitarius slices

Mark W. Doyle, Timothy W. Bailey, Young Ho Jin, Suzanne M. Appleyard, Malcolm J. Low, Michael Andresen

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The indistinct regional anatomy and intermixing of second order neurons with projection and interneurons make cellular studies more difficult within the nucleus tractus solitarius (NTS). Here, we outline experimental strategies to join in vitro electrophysiological with neuroanatomical protocols to discriminate specific subpopulations of NTS neurons. Horizontally cutting the brain stem produces slices in which electrical activation of the solitary tract (ST) is free of local interneuron contamination. Such ST excitatory synaptic currents (EPSCs) functionally identify second order NTS neurons by their minimal variation of latency (jitter). Sapphire blades, cold cutting temperatures and a mechanically stable microtome were critical to consistently obtain viable slices that were optimized for infrared and fluorescence microscopy. Anterogradely transported carbocyanine dye implanted on the aortic depressor nerve anatomically identified second order NTS neurons and their ST synaptic performance conformed to the minimal jitter signature of second order neurons. Retrograde tracers and green fluorescent protein labeled neurons afford two additional promising approaches for discriminating NTS neuron phenotypes in broader system contexts. Detailed methods and troubleshooting are described. Coupling tracing techniques with electrophysiology adds important new dimensions to NTS studies and such strategies provide bridging information between cellular mechanisms, neuroanatomy and systems integration.

Original languageEnglish (US)
Pages (from-to)37-48
Number of pages12
JournalJournal of Neuroscience Methods
Volume137
Issue number1
DOIs
StatePublished - Aug 15 2004

Fingerprint

Solitary Nucleus
Neurons
Interneurons
Regional Anatomy
Carbocyanines
Systems Integration
Neuroanatomy
Aluminum Oxide
Electrophysiology
Green Fluorescent Proteins
Fluorescence Microscopy
Brain Stem
Coloring Agents
Phenotype

Keywords

  • Autonomic
  • Brainslice
  • Carbocyanine
  • Electrophysiology
  • Fluorescence
  • Retrograde tracers
  • Sensory
  • Visceral

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Strategies for cellular identification in nucleus tractus solitarius slices. / Doyle, Mark W.; Bailey, Timothy W.; Jin, Young Ho; Appleyard, Suzanne M.; Low, Malcolm J.; Andresen, Michael.

In: Journal of Neuroscience Methods, Vol. 137, No. 1, 15.08.2004, p. 37-48.

Research output: Contribution to journalArticle

Doyle, Mark W. ; Bailey, Timothy W. ; Jin, Young Ho ; Appleyard, Suzanne M. ; Low, Malcolm J. ; Andresen, Michael. / Strategies for cellular identification in nucleus tractus solitarius slices. In: Journal of Neuroscience Methods. 2004 ; Vol. 137, No. 1. pp. 37-48.
@article{8d45a6b4be844ff9a6444e257483e501,
title = "Strategies for cellular identification in nucleus tractus solitarius slices",
abstract = "The indistinct regional anatomy and intermixing of second order neurons with projection and interneurons make cellular studies more difficult within the nucleus tractus solitarius (NTS). Here, we outline experimental strategies to join in vitro electrophysiological with neuroanatomical protocols to discriminate specific subpopulations of NTS neurons. Horizontally cutting the brain stem produces slices in which electrical activation of the solitary tract (ST) is free of local interneuron contamination. Such ST excitatory synaptic currents (EPSCs) functionally identify second order NTS neurons by their minimal variation of latency (jitter). Sapphire blades, cold cutting temperatures and a mechanically stable microtome were critical to consistently obtain viable slices that were optimized for infrared and fluorescence microscopy. Anterogradely transported carbocyanine dye implanted on the aortic depressor nerve anatomically identified second order NTS neurons and their ST synaptic performance conformed to the minimal jitter signature of second order neurons. Retrograde tracers and green fluorescent protein labeled neurons afford two additional promising approaches for discriminating NTS neuron phenotypes in broader system contexts. Detailed methods and troubleshooting are described. Coupling tracing techniques with electrophysiology adds important new dimensions to NTS studies and such strategies provide bridging information between cellular mechanisms, neuroanatomy and systems integration.",
keywords = "Autonomic, Brainslice, Carbocyanine, Electrophysiology, Fluorescence, Retrograde tracers, Sensory, Visceral",
author = "Doyle, {Mark W.} and Bailey, {Timothy W.} and Jin, {Young Ho} and Appleyard, {Suzanne M.} and Low, {Malcolm J.} and Michael Andresen",
year = "2004",
month = "8",
day = "15",
doi = "10.1016/j.jneumeth.2004.02.007",
language = "English (US)",
volume = "137",
pages = "37--48",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Strategies for cellular identification in nucleus tractus solitarius slices

AU - Doyle, Mark W.

AU - Bailey, Timothy W.

AU - Jin, Young Ho

AU - Appleyard, Suzanne M.

AU - Low, Malcolm J.

AU - Andresen, Michael

PY - 2004/8/15

Y1 - 2004/8/15

N2 - The indistinct regional anatomy and intermixing of second order neurons with projection and interneurons make cellular studies more difficult within the nucleus tractus solitarius (NTS). Here, we outline experimental strategies to join in vitro electrophysiological with neuroanatomical protocols to discriminate specific subpopulations of NTS neurons. Horizontally cutting the brain stem produces slices in which electrical activation of the solitary tract (ST) is free of local interneuron contamination. Such ST excitatory synaptic currents (EPSCs) functionally identify second order NTS neurons by their minimal variation of latency (jitter). Sapphire blades, cold cutting temperatures and a mechanically stable microtome were critical to consistently obtain viable slices that were optimized for infrared and fluorescence microscopy. Anterogradely transported carbocyanine dye implanted on the aortic depressor nerve anatomically identified second order NTS neurons and their ST synaptic performance conformed to the minimal jitter signature of second order neurons. Retrograde tracers and green fluorescent protein labeled neurons afford two additional promising approaches for discriminating NTS neuron phenotypes in broader system contexts. Detailed methods and troubleshooting are described. Coupling tracing techniques with electrophysiology adds important new dimensions to NTS studies and such strategies provide bridging information between cellular mechanisms, neuroanatomy and systems integration.

AB - The indistinct regional anatomy and intermixing of second order neurons with projection and interneurons make cellular studies more difficult within the nucleus tractus solitarius (NTS). Here, we outline experimental strategies to join in vitro electrophysiological with neuroanatomical protocols to discriminate specific subpopulations of NTS neurons. Horizontally cutting the brain stem produces slices in which electrical activation of the solitary tract (ST) is free of local interneuron contamination. Such ST excitatory synaptic currents (EPSCs) functionally identify second order NTS neurons by their minimal variation of latency (jitter). Sapphire blades, cold cutting temperatures and a mechanically stable microtome were critical to consistently obtain viable slices that were optimized for infrared and fluorescence microscopy. Anterogradely transported carbocyanine dye implanted on the aortic depressor nerve anatomically identified second order NTS neurons and their ST synaptic performance conformed to the minimal jitter signature of second order neurons. Retrograde tracers and green fluorescent protein labeled neurons afford two additional promising approaches for discriminating NTS neuron phenotypes in broader system contexts. Detailed methods and troubleshooting are described. Coupling tracing techniques with electrophysiology adds important new dimensions to NTS studies and such strategies provide bridging information between cellular mechanisms, neuroanatomy and systems integration.

KW - Autonomic

KW - Brainslice

KW - Carbocyanine

KW - Electrophysiology

KW - Fluorescence

KW - Retrograde tracers

KW - Sensory

KW - Visceral

UR - http://www.scopus.com/inward/record.url?scp=2442707205&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442707205&partnerID=8YFLogxK

U2 - 10.1016/j.jneumeth.2004.02.007

DO - 10.1016/j.jneumeth.2004.02.007

M3 - Article

C2 - 15196825

AN - SCOPUS:2442707205

VL - 137

SP - 37

EP - 48

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 1

ER -