Deletion of Tsc2 in Nociceptors Reduces Target Innervation, Ion Channel Expression, and Sensitivity to Heat.
The mechanistic target of rapamycin complex 1 (mTORC1) is known to regulate cellular growth pathways, and its genetic activation is sufficient to enhance regenerative axon growth following injury to the central or peripheral nervous systems. However, excess mTORC1 activation may promote innervation defects, and mTORC1 activity mediates injury-induced hypersensitivity, reducing enthusiasm for the pathway as a therapeutic target. While mTORC1 activity is required for full expression of some pain modalities, the effects of pathway activation on nociceptor phenotypes and sensory behaviors are currently unknown. To address this, we genetically activated mTORC1 in mouse peripheral sensory neurons by conditional deletion of its negative regulator Tuberous Sclerosis Complex 2 (Tsc2). Consistent with the well-known role of mTORC1 in regulating cell size, soma size and axon diameter of C-nociceptors were increased in Tsc2-deleted mice. Glabrous skin and spinal cord innervation by C-fiber neurons were also disrupted. Transcriptional profiling of nociceptors enriched by fluorescence-associated cell sorting (FACS) revealed downregulation of multiple classes of ion channels as well as reduced expression of markers for peptidergic nociceptors in Tsc2-deleted mice. In addition to these changes in innervation and gene expression, Tsc2-deleted mice exhibited reduced noxious heat sensitivity and decreased injury-induced cold hypersensitivity, but normal baseline sensitivity to cold and mechanical stimuli. Together, these data show that excess mTORC1 activity in sensory neurons produces changes in gene expression, neuron morphology and sensory behavior.
Pubmed ID: 29766046 RIS Download
Research resources used in this publication
- Fiji (RRID:SCR_002285) (software resource)
- ImageJ (RRID:SCR_003070) (software resource)
- B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J (RRID:IMSR_JAX:007906) (organism)
- Tsc2tm1Djk/Tsc2tm1Djk (RRID:MGI:2174787) (organism)
- Tsc2tm1.1Mjg/Tsc2tm1.1Mjg (RRID:MGI:3712786) (organism)
- Substance P antibody (RRID:AB_297089) (antibody)
- Anti-TrkA (RRID:AB_310180) (antibody)
- GOAT ANTI RAT CALCITONIN GENE-RELATED PEPTIDE (RRID:AB_2290729) (antibody)
- Neuron specific beta III Tubulin antibody (RRID:AB_10899689) (antibody)
- Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb (RRID:AB_2269803) (antibody)
- Rabbit Anti-p70 S6 Kinase Monoclonal Antibody, Unconjugated, Clone 49D7 (RRID:AB_390722) (antibody)
- Anti-alpha Tubulin antibody - Microtubule Marker (RRID:AB_2210057) (antibody)
- B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J (RRID:IMSR_JAX:007906) (organism)
- Tsc2tm1Djk/Tsc2tm1Djk (RRID:MGI:2174787) (organism)
- Tsc2tm1.1Mjg/Tsc2tm1.1Mjg (RRID:MGI:3712786) (organism)
- Anti-Neurofilament 200 kDa, clone N52 (RRID:AB_2149763) (antibody)
- Rabbit Anti-Tubulin, beta, Class III, Neuronal Polyclonal Antibody, Unconjugated (RRID:AB_291637) (antibody)
Additional research tools detected in this publication
Antibodies used in this publication
- Anti-Neurofilament 200 kDa, clone N52 (RRID:AB_2149763)
- Purified anti-Tubulin beta-3 (TUBB3) (RRID:AB_2564645)
- Substance P antibody (RRID:AB_297089)
- Anti-TrkA (RRID:AB_310180)
- GOAT ANTI RAT CALCITONIN GENE-RELATED PEPTIDE (RRID:AB_2290729)
- Neuron specific beta III Tubulin antibody (RRID:AB_10899689)
- Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb (RRID:AB_2269803)
- Rabbit Anti-p70 S6 Kinase Monoclonal Antibody, Unconjugated, Clone 49D7 (RRID:AB_390722)
- Anti-alpha Tubulin antibody - Microtubule Marker (RRID:AB_2210057)
- Rabbit Anti-Tubulin, beta, Class III, Neuronal Polyclonal Antibody, Unconjugated (RRID:AB_291637)
Associated grants
- Agency: NIDCR NIH HHS, United States
Id: R01 DE022000 - Agency: NINDS NIH HHS, United States
Id: R21 NS099603
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BioLegend (tool)
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Commercial antibody supplier and developer for biomedical research. These products are compatible with use in flow cytometry and mass cytometry, immunoprecipitation and chip, western blotting, immunofluorescence microscopy, and quantitative multiplexing.
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View all literature mentionsPrimerBank (tool)
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Database of human and mouse primer pairs for gene expression analysis by polymerase chain reaction (PCR) and quantitative PCR (qPCR). A total of 306,800 primers covering most known human and mouse genes can be accessed from the PrimerBank database, together with information on these primers such as T(m), location on the transcript and amplicon size. For each gene, at least one primer pair has been designed and in many cases alternative primer pairs exist. Primers have been designed to work under the same PCR conditions, thus facilitating high-throughput QPCR. All primers in PrimerBank were carefully designed to ensure gene specificity. All experimental validation data for mouse primers are available from PrimerBank. You can submit your primers. They will be added to the database once they are properly QCd.
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THIS RESOURCE IS NO LONGER IN SERVICE. Documented on March 17, 2022. An integrated software suite for functional analysis of experimental data. The scope of data types includes microarray and SAGE gene expression, SNPs and CGH arrays, proteomics, metabolomics, pathway analysis, Y2H and other custom interactions. MetaCore is based on a proprietary manually curated database of human protein-protein, protein-DNA and protein compound interactions, metabolic and signaling pathways and the effects of bioactive molecules in gene expression.
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Script distributed with the HT-Seq Python framework for processing RNA-seq or DNA-seq data.
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Quality control software that perform checks on raw sequence data coming from high throughput sequencing pipelines. This software also provides a modular set of analyses which can give a quick impression of the quality of the data prior to further analysis.
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Software package for differential gene expression analysis based on the negative binomial distribution. Used for analyzing RNA-seq data for differential analysis of count data, using shrinkage estimation for dispersions and fold changes to improve stability and interpretability of estimates.
View all literature mentionsAnti-Neurofilament 200 kDa, clone N52 (antibody)
RRID:AB_2149763
This monoclonal targets Neurofilament 200 kD
View all literature mentionsPurified anti-Tubulin beta-3 (TUBB3) (antibody)
RRID:AB_2564645
This unknown targets Tubulin beta-3
View all literature mentionsSubstance P antibody (antibody)
RRID:AB_297089
This polyclonal targets Substance P
View all literature mentionsGOAT ANTI RAT CALCITONIN GENE-RELATED PEPTIDE (antibody)
RRID:AB_2290729
This polyclonal targets GOAT ANTI RAT CALCITONIN GENE-RELATED PEPTIDE
View all literature mentionsNeuron specific beta III Tubulin antibody (antibody)
RRID:AB_10899689
This polyclonal targets Neuron specific beta III Tubulin antibody
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RRID:AB_2269803
This monoclonal targets p70 S6 Kinase, phospho (Thr389)
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RRID:AB_390722
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This polyclonal targets alpha Tubulin
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RRID:AB_291637
This polyclonal targets Tubulin, beta, Class III, Neuronal
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Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.
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Open source Java based image processing software program designed for scientific multidimensional images. ImageJ has been transformed to ImageJ2 application to improve data engine to be sufficient to analyze modern datasets.
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RRID:IMSR_JAX:007906
Mus musculus with name B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J from IMSR.
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RRID:MGI:2174787
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RRID:MGI:3712786
Allele Detail: Targeted This is a legacy resource.
View all literature mentionsSubstance P antibody (antibody)
RRID:AB_297089
This polyclonal targets Substance P
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RRID:AB_2290729
This polyclonal targets GOAT ANTI RAT CALCITONIN GENE-RELATED PEPTIDE
View all literature mentionsNeuron specific beta III Tubulin antibody (antibody)
RRID:AB_10899689
This polyclonal targets Neuron specific beta III Tubulin antibody
View all literature mentionsPhospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb (antibody)
RRID:AB_2269803
This monoclonal targets p70 S6 Kinase, phospho (Thr389)
View all literature mentionsRabbit Anti-p70 S6 Kinase Monoclonal Antibody, Unconjugated, Clone 49D7 (antibody)
RRID:AB_390722
This monoclonal targets p70 S6 Kinase
View all literature mentionsAnti-alpha Tubulin antibody - Microtubule Marker (antibody)
RRID:AB_2210057
This polyclonal targets alpha Tubulin
View all literature mentionsB6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J (organism)
RRID:IMSR_JAX:007906
Mus musculus with name B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J from IMSR.
View all literature mentionsTsc2tm1Djk/Tsc2tm1Djk (organism)
RRID:MGI:2174787
Allele Detail: Targeted This is a legacy resource.
View all literature mentionsTsc2tm1.1Mjg/Tsc2tm1.1Mjg (organism)
RRID:MGI:3712786
Allele Detail: Targeted This is a legacy resource.
View all literature mentionsAnti-Neurofilament 200 kDa, clone N52 (antibody)
RRID:AB_2149763
This monoclonal targets Neurofilament 200 kD
View all literature mentionsRabbit Anti-Tubulin, beta, Class III, Neuronal Polyclonal Antibody, Unconjugated (antibody)
RRID:AB_291637
This polyclonal targets Tubulin, beta, Class III, Neuronal
View all literature mentions
