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ScRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals maturational differences in gene expression

ABSTRACT: Cultured dorsal root ganglia neurons of 4 and 12 weeks old C57Bl/6 mice were first characterized as mechanosensitive (showed rapidly adapting mechanically actived currents in whole-cell votage clamp) and as mechanoreceptors (evoked action potential waveform in whole-cell current clamp). Next, cells were subjected to RNA-seq and sequences were analyzed bioinformatically. We found maturatinal changes comprising age-dependent differential gene expression in these somatosensory mechanoreceptors. The found differences call for tight age-matching of young mice in research and underline found electrophysiological and behavioural maturational changes in somatosensation, see Michel, Niklas and Narayanan, Pratibha and Shomroni, Orr and Schmidt, Manuela, Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood. CELL-REPORTS-D-20-00425. Available at SSRN: https://ssrn.com/abstract=3536401 or http://dx.doi.org/10.2139/ssrn.3536401

ORGANISM(S): Mus musculus

PROVIDER: GSE154159 | GEO | 2020/07/21

REPOSITORIES: GEO

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scRNA-seq of electrophysiologically characterized DRG neurons reveals subtype-specific and maturational differences in gene expression

Project description:Cultured dorsal root ganglia neurons of 4 and 12 weeks old C57Bl/6 mice were first characterized as mechanosensitive (showed rapidly adapting mechanically actived currents in whole-cell votage clamp or not (non-responders)) and as either mechanoreceptors or nociceptors (evoked action potential waveform in whole-cell current clamp). Next, cells were subjected to RNA-seq and sequences were analyzed bioinformatically. We found maturatinal changes comprising subtype-specific and age-dependent differential gene expression in these somatosensory neurons. See Michel, Niklas and Narayanan, Pratibha and Shomroni, Orr and Schmidt, Manuela, Cutaneous Touch and Piezo2-Mediated Mechanotransduction Decline Throughout Mouse Maturation Until Adulthood. CELL-REPORTS-D-20-00425. Available at SSRN: https://ssrn.com/abstract=3536401 or http://dx.doi.org/10.2139/ssrn.3536401

2020-07-21 | GSE154160 | GEO

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MicroRNA-183 cluster continuously scales mechanical pain sensitivity by regulating basal and neuropathic pain gene pathways.

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Project description:Cutaneous C-unmyelinated free nerve endings and hair follicles-innervating C-LTMRs convey two opposite aspects of touch sensation: a sensation of pain and a sensation of pleasant touch. The molecular mechanisms underlying these diametrically opposite functions are unknown. Here we used a mouse model that genetically marks C-LTMRs and the free nerve endings MRGPRD+ neurons in combination with fluorescent cell surface labeling, flow cytometry and RNA deep-sequencing technology. Cluster analysis of the RNA-Seq profiles of the purified neuronal subsets revealed 156 and 184 genes differentially expressed in MRGPRD-expressing neurons and C-LTMRs, respectively. 48 MRGPD- and 67 C-LTMRs-enriched genes were validated using a triple staining experiment approach and the Cav3.3 channel, found to be exclusively expressed in C-LTMRs, was validated using electrophysiology. Furthermore, our study greatly expands the molecular characterization of C-LTMRs and suggests that this particular population of neurons shares common transcriptional signatures with A and A low threshold mechanoreceptors. RNA profiles of FACS-sorted subsets of sensory neurons, namely Mrgprd+ neurons (DP, double-positive=IB4+GINIP+), C-LTMRs (IB4-GINIP+) and remaining cells (DN, double-negative=IB4-GINIP-) were generated by deep sequencing in duplicate using Illumina HiSeq 2000.

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Transcriptional profiling of cutaneous Mrgprd free nerve endings and C-LTMRs

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