Universität zu Köln

Isensee, Joerg ORCID: 0000-0002-3390-0051, Wenzel, Carsten, Buschow, Rene ORCID: 0000-0002-9800-2578, Weissmann, Robert, Kuss, Andreas W. and Hucho, Tim ORCID: 0000-0002-4147-9308 (2014). Subgroup-Elimination Transcriptomics Identifies Signaling Proteins that Define Subclasses of TRPV1-Positive Neurons and a Novel Paracrine Circuit. PLoS One, 9 (12). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

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Abstract

Normal and painful stimuli are detected by specialized subgroups of peripheral sensory neurons. The understanding of the functional differences of each neuronal subgroup would be strongly enhanced by knowledge of the respective subgroup transcriptome. The separation of the subgroup of interest, however, has proven challenging as they can hardly be enriched. Instead of enriching, we now rapidly eliminated the subgroup of neurons expressing the heat-gated cation channel TRPV1 from dissociated rat sensory ganglia. Elimination was accomplished by brief treatment with TRPV1 agonists followed by the removal of compromised TRPV1(+) neurons using density centrifugation. By differential microarray and sequencing (RNA-Seq) based expression profiling we compared the transcriptome of all cells within sensory ganglia versus the same cells lacking TRPV1 expressing neurons, which revealed 240 differentially expressed genes (adj. p<.05, fold-change >1.5). Corroborating the specificity of the approach, many of these genes have been reported to be involved in noxious heat or pain sensitization. Beyond the expected enrichment of ion channels, we found the TRPV1 transcriptome to be enriched for GPCRs and other signaling proteins involved in adenosine, calcium, and phosphatidylinositol signaling. Quantitative population analysis using a recent High Content Screening (HCS) microscopy approach identified substantial heterogeneity of expressed target proteins even within TRPV1-positive neurons. Signaling components defined distinct further subgroups within the population of TRPV1-positive neurons. Analysis of one such signaling system showed that the pain sensitizing prostaglandin PGD2 activates DP1 receptors expressed predominantly on TRPV1(+) neurons. In contrast, we found the PGD2 producing prostaglandin D synthase to be expressed exclusively in myelinated large-diameter neurons lacking TRPV1, which suggests a novel paracrine neuron-neuron communication. Thus, subgroup analysis based on the elimination rather than enrichment of the subgroup of interest revealed proteins that define subclasses of TRPV1-positive neurons and suggests a novel paracrine circuit.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Isensee, Joerg UNSPECIFIED orcid.org/0000-0002-3390-0051 UNSPECIFIED Wenzel, Carsten UNSPECIFIED UNSPECIFIED UNSPECIFIED Buschow, Rene UNSPECIFIED orcid.org/0000-0002-9800-2578 UNSPECIFIED Weissmann, Robert UNSPECIFIED UNSPECIFIED UNSPECIFIED Kuss, Andreas W. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hucho, Tim UNSPECIFIED orcid.org/0000-0002-4147-9308 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-419910
DOI:	10.1371/journal.pone.0115731
Journal or Publication Title:	PLoS One
Volume:	9
Number:	12
Date:	2014
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DORSAL-ROOT GANGLION; GENE-RELATED PEPTIDE; PROSTATIC ACID-PHOSPHATASE; PERIPHERAL SENSORY NEURONS; PRIMARY AFFERENT NEURONS; CAPSAICIN RECEPTOR; INFLAMMATORY PAIN; KINASE-II; VANILLOID RECEPTOR-1; MOLECULAR-MECHANISMS Multiple languages Multidisciplinary Sciences Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/41991