Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices

Andrei V. Derbenev; Hanad Duale; Alexander G. Rabchevsky; Bret N. Smith

doi:10.1016/j.neulet.2010.03.036

Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices

Andrei V. Derbenev, Hanad Duale, Alexander G. Rabchevsky, Bret N. Smith

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Abstract

Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was -51±4mV and input resistance was 448±39MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1±1Hz) was approximately 2.5 times that for sIPSCs (1.4±0.3Hz). Application of the glycine and GABA_A receptor-linked Cl^- channel blocker, picrotoxin (100μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.

Original language	English
Pages (from-to)	168-172
Number of pages	5
Journal	Neuroscience Letters
Volume	474
Issue number	3
DOIs	https://doi.org/10.1016/j.neulet.2010.03.036
State	Published - May 2010

Bibliographical note

Funding Information:
We thank Dr. Lynn Enquist (Princeton University) for the PRV-152. Funded by PVA grant # 2561 (H. Duale), NIHLBI , 1R21HL091293 (A.V. Derbenev), NINDS , R01 NS049901 (A.G. Rabchevsky), NIDDK , R01 DK056132 (B.N. Smith), and NINDS , P30 NS051220 .

Keywords

Intermediolateral cell column
Preganglionic neuron
Pseudorabies virus
Sympathetic
Whole-cell patch-clamp

ASJC Scopus subject areas

General Neuroscience

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10.1016/j.neulet.2010.03.036

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title = "Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices",

abstract = "Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was -51±4mV and input resistance was 448±39MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1±1Hz) was approximately 2.5 times that for sIPSCs (1.4±0.3Hz). Application of the glycine and GABAA receptor-linked Cl- channel blocker, picrotoxin (100μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.",

keywords = "Intermediolateral cell column, Preganglionic neuron, Pseudorabies virus, Sympathetic, Whole-cell patch-clamp",

author = "Derbenev, {Andrei V.} and Hanad Duale and Rabchevsky, {Alexander G.} and Smith, {Bret N.}",

note = "Funding Information: We thank Dr. Lynn Enquist (Princeton University) for the PRV-152. Funded by PVA grant # 2561 (H. Duale), NIHLBI , 1R21HL091293 (A.V. Derbenev), NINDS , R01 NS049901 (A.G. Rabchevsky), NIDDK , R01 DK056132 (B.N. Smith), and NINDS , P30 NS051220 .",

year = "2010",

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doi = "10.1016/j.neulet.2010.03.036",

language = "English",

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T1 - Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices

AU - Derbenev, Andrei V.

AU - Duale, Hanad

AU - Rabchevsky, Alexander G.

AU - Smith, Bret N.

N1 - Funding Information: We thank Dr. Lynn Enquist (Princeton University) for the PRV-152. Funded by PVA grant # 2561 (H. Duale), NIHLBI , 1R21HL091293 (A.V. Derbenev), NINDS , R01 NS049901 (A.G. Rabchevsky), NIDDK , R01 DK056132 (B.N. Smith), and NINDS , P30 NS051220 .

PY - 2010/5

Y1 - 2010/5

N2 - Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was -51±4mV and input resistance was 448±39MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1±1Hz) was approximately 2.5 times that for sIPSCs (1.4±0.3Hz). Application of the glycine and GABAA receptor-linked Cl- channel blocker, picrotoxin (100μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.

AB - Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was -51±4mV and input resistance was 448±39MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1±1Hz) was approximately 2.5 times that for sIPSCs (1.4±0.3Hz). Application of the glycine and GABAA receptor-linked Cl- channel blocker, picrotoxin (100μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.

KW - Intermediolateral cell column

KW - Preganglionic neuron

KW - Pseudorabies virus

KW - Sympathetic

KW - Whole-cell patch-clamp

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JO - Neuroscience Letters

JF - Neuroscience Letters

IS - 3

ER -

Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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