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Mouse DRG Cell Line with Properties of Nociceptors

In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry...

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Autores principales: Doran, Ciara, Chetrit, Jonathan, Holley, Matthew C., Grundy, David, Nassar, Mohammed A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460156/
https://www.ncbi.nlm.nih.gov/pubmed/26053673
http://dx.doi.org/10.1371/journal.pone.0128670
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author Doran, Ciara
Chetrit, Jonathan
Holley, Matthew C.
Grundy, David
Nassar, Mohammed A.
author_facet Doran, Ciara
Chetrit, Jonathan
Holley, Matthew C.
Grundy, David
Nassar, Mohammed A.
author_sort Doran, Ciara
collection PubMed
description In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry, RT-PCR and calcium microfluorimetry, we demonstrate that the cell line MED17.11 expresses markers of cells committed to the sensory neuron lineage. Within a few hours under differentiating conditions, MED17.11 cells extend processes and following seven days of differentiation, express markers of more mature DRG neurons, such as NaV1.7 and Piezo2. However, at least at this time-point, the nociceptive marker NaV1.8 is not expressed, but the cells respond to compounds known to excite nociceptors, including the TRPV1 agonist capsaicin, the purinergic receptor agonist ATP and the voltage gated sodium channel agonist, veratridine. Robust calcium transients are observed in the presence of the inflammatory mediators bradykinin, histamine and norepinephrine. MED17.11 cells have the potential to replace or reduce the use of primary DRG culture in sensory, pain and developmental research by providing a simple model to study acute nociception, neurite outgrowth and the developmental specification of DRG neurons.
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spelling pubmed-44601562015-06-16 Mouse DRG Cell Line with Properties of Nociceptors Doran, Ciara Chetrit, Jonathan Holley, Matthew C. Grundy, David Nassar, Mohammed A. PLoS One Research Article In vitro cell lines from DRG neurons aid drug discovery because they can be used for early stage, high-throughput screens for drugs targeting pain pathways, with minimal dependence on animals. We have established a conditionally immortal DRG cell line from the Immortomouse. Using immunocytochemistry, RT-PCR and calcium microfluorimetry, we demonstrate that the cell line MED17.11 expresses markers of cells committed to the sensory neuron lineage. Within a few hours under differentiating conditions, MED17.11 cells extend processes and following seven days of differentiation, express markers of more mature DRG neurons, such as NaV1.7 and Piezo2. However, at least at this time-point, the nociceptive marker NaV1.8 is not expressed, but the cells respond to compounds known to excite nociceptors, including the TRPV1 agonist capsaicin, the purinergic receptor agonist ATP and the voltage gated sodium channel agonist, veratridine. Robust calcium transients are observed in the presence of the inflammatory mediators bradykinin, histamine and norepinephrine. MED17.11 cells have the potential to replace or reduce the use of primary DRG culture in sensory, pain and developmental research by providing a simple model to study acute nociception, neurite outgrowth and the developmental specification of DRG neurons. Public Library of Science 2015-06-08 /pmc/articles/PMC4460156/ /pubmed/26053673 http://dx.doi.org/10.1371/journal.pone.0128670 Text en © 2015 Doran et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Doran, Ciara
Chetrit, Jonathan
Holley, Matthew C.
Grundy, David
Nassar, Mohammed A.
Mouse DRG Cell Line with Properties of Nociceptors
title Mouse DRG Cell Line with Properties of Nociceptors
title_full Mouse DRG Cell Line with Properties of Nociceptors
title_fullStr Mouse DRG Cell Line with Properties of Nociceptors
title_full_unstemmed Mouse DRG Cell Line with Properties of Nociceptors
title_short Mouse DRG Cell Line with Properties of Nociceptors
title_sort mouse drg cell line with properties of nociceptors
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4460156/
https://www.ncbi.nlm.nih.gov/pubmed/26053673
http://dx.doi.org/10.1371/journal.pone.0128670
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