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ScRNA-seq of electrophysiologically characterized DRG neurons reveals subtype-specific and 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 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

ORGANISM(S): Mus musculus

PROVIDER: GSE154160 | GEO | 2020/07/21

REPOSITORIES: GEO

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scRNA-seq of electrophysiologically characterized mechanosensetive DRG neurons reveals 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) 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

2020-07-21 | GSE154159 | GEO

RNASeq of trigeminal ganglia of chicken and wild duck species

Project description:A major challenge in biology is to link cellular and molecular variations with behavioral phenotypes. Here, we approached this by studying somatosensory neurons from a panel of bird species from the family Anatidae, known for their tactile-based foraging behavior. We found that tactile specialists exhibit a proportional expansion of neuronal mechanoreceptors in trigeminal ganglia. The expansion of mechanoreceptors occurs via neurons with intermediately and slowly inactivating mechano-current. Such neurons contain the Piezo2 ion channel, whose expression positively correlates with the expression of factors responsible for the development and function of touch receptors. Conversely, Piezo2 expression negatively correlates with expression of molecules mediating the detection of temperature and pain, suggesting that the expansion of Piezo2-containing mechanoreceptors with prolonged mechano-current occurs at the expense of other types of sensory neurons. Our study reveals a general mechanism of tactile specialization in vertebrates at the level of somatosensory system.

2019-01-29 | GSE125754 | GEO

Maturational changes in mouse cutaneous touch and Piezo2-mediated mechanotransduction

Project description:Maturational changes in mouse cutaneous touch and Piezo2-mediated mechanotransduction

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Project description:Nociception is protective and prevents tissue damage but can also facilitate chronic pain. If a general principle governs these two types of pain is unknown. Here, we show that both basal mechanical and neuropathic pain are controlled by microRNA-183 cluster in mice. This single cluster controls more than 80% of neuropathic pain-regulated genes and scales basal mechanical sensitivity and mechanical allodynia by regulating auxiliary voltage-gated calcium channel subunits a2d. Basal sensitivity is controlled in nociceptors and allodynia involves TrkB+ light-touch mechanoreceptors. These light-touch sensitive neurons that normally do not elicit pain produce pain during neuropathy that is reversed by gabapentin. Thus, a single miRNA cluster continuously scales acute noxious mechanical sensitivity in nociceptive neurons and suppresses neuropathic pain transduction in a specific, light-touch sensitive neuronal type recruited during mechanical allodynia.

2017-05-25 | GSE99265 | GEO

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

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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Project description:Mechanosensory neurons innervating the skin underlie our sense of touch. Fast-conducting, rapidly adapting mechanoreceptors innervating glabrous (non-hairy) skin form Meissner corpuscles, while in hairy skin, they associate with hair follicles, forming longitudinal lanceolate endings. How mechanoreceptors develop axonal endings appropriate for their skin targets is unknown. We report that mechanoreceptor morphologies across different skin regions are indistinguishable during early development but diverge post-natally, in parallel with skin maturation. Neurons terminating along the glabrous and hairy skin border exhibit hybrid morphologies, forming both Meissner corpuscles and lanceolate endings. Additionally, molecular profiles of neonatal glabrous and hairy skin-innervating neurons largely overlap. In mouse mutants with ectopic glabrous skin, mechanosensory neurons form end-organs appropriate for the altered skin type. Finally, BMP5 and BMP7 are enriched in glabrous skin, and signaling through type I bone morphogenetic protein (BMP) receptors in neurons is critical for Meissner corpuscle morphology. Thus, mechanoreceptor morphogenesis is flexibly instructed by target tissues.

2023-07-27 | GSE229111 | GEO

Genome-wide analysis of P15 dorsal root ganglion gene expression of control and Isl1-Cre//c-Maf mutant mice

Project description:The sense of touch relies on the detection of mechanical stimuli by specialized sensory neurons. Mechanosensory neurons are generated from neural crest cells, but the scarcity of molecular data has made it difficult to analyze their development and to define the basis of their diversity and function. The transcription factor c-Maf is crucial for mechanosensory function in mice and humans. In particular, c-Maf is required for the development and function of mechanosensory neurons terminating in lanceolate endings, Meissner corpuscles and Pacinian corpuscles. c-Maf is a key transcription factor directing the development and function of rapidly-adapting mechanoreceptors and their end organs.

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

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A vagal reflex evoked by airway closure

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Project description:In vitro generation of human peripheral sensory neurons may provide a framework for novel drug screening platforms and disease models of touch and pain. However, derivation of a functionally pure sensory neuron population remains a major unmet challenge. We discovered that, by expressing the transcription factors NGN2 and BRN3A, human pluripotent stem cells can be induced to differentiate into a surprisingly homogenous culture of cold- and mechano-sensing neurons. Although such a neuronal subtype has not been reported in mice, we found molecular evidence of its existence in adult human sensory ganglia. Combining NGN2 and BRN3A programming with neural crest patterning, we produced two additional populations of sensory neurons, including a more specialized mechanosensory neuron subtype. Finally, we applied this system to model a rare inherited sensory disorder, characterized by profound impairment of touch sensation and proprioception, caused by inactivating mutations in PIEZO2. Together these findings establish an approach to specify distinct sensory neuron subtypes in vitro, underscoring the utility of stem cell technology to capture human-specific features of physiology and disease.

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