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A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia

BACKGROUND: The cell bodies of sensory neurons, which transmit information from the external environment to the spinal cord, can be found at all levels of the spinal column in paired structures called dorsal root ganglia (DRG). Rodent DRG neurons have long been studied in the laboratory to improve u...

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Autores principales: Sleigh, James N., Weir, Greg A., Schiavo, Giampietro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750296/
https://www.ncbi.nlm.nih.gov/pubmed/26864470
http://dx.doi.org/10.1186/s13104-016-1915-8
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author Sleigh, James N.
Weir, Greg A.
Schiavo, Giampietro
author_facet Sleigh, James N.
Weir, Greg A.
Schiavo, Giampietro
author_sort Sleigh, James N.
collection PubMed
description BACKGROUND: The cell bodies of sensory neurons, which transmit information from the external environment to the spinal cord, can be found at all levels of the spinal column in paired structures called dorsal root ganglia (DRG). Rodent DRG neurons have long been studied in the laboratory to improve understanding of sensory nerve development and function, and have been instrumental in determining mechanisms underlying pain and neurodegeneration in disorders of the peripheral nervous system. Here, we describe a simple, step-by-step protocol for the swift isolation of mouse DRG, which can be enzymatically dissociated to produce fully differentiated primary neuronal cultures, or processed for downstream analyses, such as immunohistochemistry or RNA profiling. FINDINGS: After dissecting out the spinal column, from the base of the skull to the level of the femurs, it can be cut down the mid-line and the spinal cord and meninges removed, before extracting the DRG and detaching unwanted axons. This protocol allows the easy and rapid isolation of DRG with minimal practice and dissection experience. The process is both faster and less technically challenging than extracting the ganglia from the in situ column after performing a dorsal laminectomy. CONCLUSIONS: This approach reduces the time required to collect DRG, thereby improving efficiency, permitting less opportunity for tissue deterioration, and, ultimately, increasing the chances of generating healthy primary DRG cultures or high quality, reproducible experiments using DRG tissue.
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spelling pubmed-47502962016-02-12 A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia Sleigh, James N. Weir, Greg A. Schiavo, Giampietro BMC Res Notes Technical Note BACKGROUND: The cell bodies of sensory neurons, which transmit information from the external environment to the spinal cord, can be found at all levels of the spinal column in paired structures called dorsal root ganglia (DRG). Rodent DRG neurons have long been studied in the laboratory to improve understanding of sensory nerve development and function, and have been instrumental in determining mechanisms underlying pain and neurodegeneration in disorders of the peripheral nervous system. Here, we describe a simple, step-by-step protocol for the swift isolation of mouse DRG, which can be enzymatically dissociated to produce fully differentiated primary neuronal cultures, or processed for downstream analyses, such as immunohistochemistry or RNA profiling. FINDINGS: After dissecting out the spinal column, from the base of the skull to the level of the femurs, it can be cut down the mid-line and the spinal cord and meninges removed, before extracting the DRG and detaching unwanted axons. This protocol allows the easy and rapid isolation of DRG with minimal practice and dissection experience. The process is both faster and less technically challenging than extracting the ganglia from the in situ column after performing a dorsal laminectomy. CONCLUSIONS: This approach reduces the time required to collect DRG, thereby improving efficiency, permitting less opportunity for tissue deterioration, and, ultimately, increasing the chances of generating healthy primary DRG cultures or high quality, reproducible experiments using DRG tissue. BioMed Central 2016-02-11 /pmc/articles/PMC4750296/ /pubmed/26864470 http://dx.doi.org/10.1186/s13104-016-1915-8 Text en © Sleigh et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Technical Note
Sleigh, James N.
Weir, Greg A.
Schiavo, Giampietro
A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title_full A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title_fullStr A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title_full_unstemmed A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title_short A simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
title_sort simple, step-by-step dissection protocol for the rapid isolation of mouse dorsal root ganglia
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750296/
https://www.ncbi.nlm.nih.gov/pubmed/26864470
http://dx.doi.org/10.1186/s13104-016-1915-8
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