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Defined Medium Organotypic Cultures of Spinal Cord Put ‘Pain in a Dish’

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Isolated Central Nervous System Circuits

Part of the book series: Neuromethods ((NM,volume 73))

Abstract

Nerve injury and/or disease are known to provoke the release of various neurotrophic and inflammatory agents from spinal microglia and astrocytes. Prolonged exposure of spinal neurons to these ‘nociceptive mediators’ invokes pathophysiological changes that lead to the ‘central sensitization’ which characterizes chronic pain. In order to investigate their actions, it is necessary to expose spinal neural networks to individual putative mediators for prolonged periods as would occur following injury or in the presence of disease. Because this is not feasible using acutely isolated spinal cord slices, we developed a defined medium organotypic spinal cord culture model which retains the structural and synaptic organization of the spinal cord in vivo. This ‘pain in a dish’ system represents an exciting new methodology for the investigation of the pathophysiological processes that underlie chronic pain.

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Abbreviations

ACSF:

Artificial cerebrospinal fluid

AdGFP:

Adenovirus containing green fluorescent protein

AM:

AcetoxyMethyl

aMCM:

Activated microglia-conditioned medium

ANOVA:

Analysis of variance assay

AraC:

Cytosine-b-D-arabinofuranoside

AUC:

Area under curve

BDNF:

Brain-derived neurotrophic factor

CCI:

Chronic constriction injury

CMV:

Cytomegalovirus

DMEM:

Dulbecco’s modified Eagle’s medium

DMOTC:

Defined medium organotypic cultures

DRG:

Dorsal root ganglion

ELISA:

Enzyme-linked immunosorbent assay

FBS:

Fetal bovine serum

GAD:

Glutamic acid decarboxylase

GcaMP:

GFP-calmodulin-protein

GFP:

Green fluorescent protein

HBSS:

Hanks buffered saline solution

IR-DIC:

Infrared differential interference contrast

Im :

Membrane current

LPS:

Lipopolysaccharide

NGF:

Nerve growth factor

OTC:

Organotypic

PBS:

Phosphate-buffered saline

ROI:

Region of interest

siRNA:

Small interfering ribonucleic acid

vGlut:

Glutamate vesicular transporter

Vm :

Membrane potential

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Acknowledgements

Supported by grants from the Canadian Institutes of Health Research, Alberta Heritage Foundation for Medical Research, Canadian Institutes for Innovation, Advanced Education Alberta and the Pfizer Canada Neuropathic Pain Research Awards Program.

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Authors and Affiliations

  1. Faculty of Medicine & Dentistry, Department of Pharmacology and Centre for Neuroscience, University of Alberta, Edmonton, AB, Canada

    James E. Biggs,  Van B. Lu, Helena J. Kim, Aaron Lai, William F. Colmers & Peter A. Smith

  2. Faculty of Medicine & Dentistry, Department of Psychiatry and Centre for Neuroscience, University of Alberta, Edmonton, AB, Canada

    Kathryn G. Todd

  3. Faculty of Medicine & Dentistry, Department of Physiology and Centre for Neuroscience, University of Alberta, Edmonton, AB, Canada

    Klaus Ballanyi

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  1. James E. Biggs
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  2. Van B. Lu
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  3. Helena J. Kim
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  4. Aaron Lai
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  5. Kathryn G. Todd
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  6. Klaus Ballanyi
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  7. William F. Colmers
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Correspondence to James E. Biggs .

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Editors and Affiliations

  1. Faculty of Medicine & Dentistry, Department of Physiology and Centre for, University of Alberta, N/A 232D, Edmonton, T6G 2S2, Alberta, Canada

    Klaus Ballanyi

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Biggs, J.E. et al. (2012). Defined Medium Organotypic Cultures of Spinal Cord Put ‘Pain in a Dish’. In: Ballanyi, K. (eds) Isolated Central Nervous System Circuits. Neuromethods, vol 73. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-020-5_14

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  • DOI: https://doi.org/10.1007/978-1-62703-020-5_14

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