Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.

Sensory neurons in dorsal root ganglia (DRG) are highly heterogeneous in terms of cell size, protein expression, and signaling activity. To analyze their heterogeneity, threshold-based methods are commonly used, which often yield highly variable results due to the subjectivity of the individual inve...

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Autores principales:	Christine Andres, Jan Hasenauer, Frank Allgower, Tim Hucho
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4e6dacbdf08048eab28aed3b285642e2
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spelling	oai:doaj.org-article:4e6dacbdf08048eab28aed3b285642e22021-11-18T07:24:11ZThreshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.1932-620310.1371/journal.pone.0034257https://doaj.org/article/4e6dacbdf08048eab28aed3b285642e22012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22479579/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Sensory neurons in dorsal root ganglia (DRG) are highly heterogeneous in terms of cell size, protein expression, and signaling activity. To analyze their heterogeneity, threshold-based methods are commonly used, which often yield highly variable results due to the subjectivity of the individual investigator. In this work, we introduce a threshold-free analysis approach for sparse and highly heterogeneous datasets obtained from cultures of sensory neurons. This approach is based on population estimates and completely free of investigator-set parameters. With a quantitative automated microscope we measured the signaling state of single DRG neurons by immunofluorescently labeling phosphorylated, i.e., activated Erk1/2. The population density of sensory neurons with and without pain-sensitizing nerve growth factor (NGF) treatment was estimated using a kernel density estimator (KDE). By subtraction of both densities and integration of the positive part, a robust estimate for the size of the responsive subpopulations was obtained. To assure sufficiently large datasets, we determined the number of cells required for reliable estimates using a bootstrapping approach. The proposed methods were employed to analyze response kinetics and response amplitude of DRG neurons after NGF stimulation. We thereby determined the portion of NGF responsive cells on a true population basis. The analysis of the dose dependent NGF response unraveled a biphasic behavior, while the study of its time dependence showed a rapid response, which approached a steady state after less than five minutes. Analyzing two parameter correlations, we found that not only the number of responsive small-sized neurons exceeds the number of responsive large-sized neurons--which is commonly reported and could be explained by the excess of small-sized cells--but also the probability that small-sized cells respond to NGF is higher. In contrast, medium-sized and large-sized neurons showed a larger response amplitude in their mean Erk1/2 activity.Christine AndresJan HasenauerFrank AllgowerTim HuchoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 3, p e34257 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christine Andres Jan Hasenauer Frank Allgower Tim Hucho Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
description	Sensory neurons in dorsal root ganglia (DRG) are highly heterogeneous in terms of cell size, protein expression, and signaling activity. To analyze their heterogeneity, threshold-based methods are commonly used, which often yield highly variable results due to the subjectivity of the individual investigator. In this work, we introduce a threshold-free analysis approach for sparse and highly heterogeneous datasets obtained from cultures of sensory neurons. This approach is based on population estimates and completely free of investigator-set parameters. With a quantitative automated microscope we measured the signaling state of single DRG neurons by immunofluorescently labeling phosphorylated, i.e., activated Erk1/2. The population density of sensory neurons with and without pain-sensitizing nerve growth factor (NGF) treatment was estimated using a kernel density estimator (KDE). By subtraction of both densities and integration of the positive part, a robust estimate for the size of the responsive subpopulations was obtained. To assure sufficiently large datasets, we determined the number of cells required for reliable estimates using a bootstrapping approach. The proposed methods were employed to analyze response kinetics and response amplitude of DRG neurons after NGF stimulation. We thereby determined the portion of NGF responsive cells on a true population basis. The analysis of the dose dependent NGF response unraveled a biphasic behavior, while the study of its time dependence showed a rapid response, which approached a steady state after less than five minutes. Analyzing two parameter correlations, we found that not only the number of responsive small-sized neurons exceeds the number of responsive large-sized neurons--which is commonly reported and could be explained by the excess of small-sized cells--but also the probability that small-sized cells respond to NGF is higher. In contrast, medium-sized and large-sized neurons showed a larger response amplitude in their mean Erk1/2 activity.
format	article
author	Christine Andres Jan Hasenauer Frank Allgower Tim Hucho
author_facet	Christine Andres Jan Hasenauer Frank Allgower Tim Hucho
author_sort	Christine Andres
title	Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
title_short	Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
title_full	Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
title_fullStr	Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
title_full_unstemmed	Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.
title_sort	threshold-free population analysis identifies larger drg neurons to respond stronger to ngf stimulation.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/4e6dacbdf08048eab28aed3b285642e2
work_keys_str_mv	AT christineandres thresholdfreepopulationanalysisidentifieslargerdrgneuronstorespondstrongertongfstimulation AT janhasenauer thresholdfreepopulationanalysisidentifieslargerdrgneuronstorespondstrongertongfstimulation AT frankallgower thresholdfreepopulationanalysisidentifieslargerdrgneuronstorespondstrongertongfstimulation AT timhucho thresholdfreepopulationanalysisidentifieslargerdrgneuronstorespondstrongertongfstimulation
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Threshold-free population analysis identifies larger DRG neurons to respond stronger to NGF stimulation.

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