Project description:Sensory hypersensitivity and somatosensory deficits represent the core symptoms of Fragile X syndrome (FXS).These alterations are believed to arise from changes in cortical sensory processing, while potential deficits in the function of peripheral sensory neurons or surrounding satellite glial cells (SGCs) residing in dorsal root ganglia remain unexplored. We found that cultured peripheral sensory neurons exhibit pronounced hyperexcitability in Fmr1 KO mice evident in markedly increased firing rate and decreased spike threshold. These alterations were caused primarily by increased input resistance, which aroused from decreased HCN channel-mediated current Ih. EM analyses also revealed major structural defects in neuron-SGC association. Single-cell RNAseq demonstrated extensive transcriptional changes in both neurons and SGCs indicative of defects in neuronal maturation/differentiation and neuron-SGC communication. These results reveal a hyper-excitable state of peripheral sensory neurons in Fmr1 KO mice with contributions from intrinsic alterations and from disrupted neuron-glia association and communication.
Project description:Peripheral sensory neurons located in dorsal root ganglia relay sensory information from the peripheral tissue to the brain. Satellite glial cells (SGC) are unique glial cells that form an envelope completely surrounding each sensory neuron soma. This organization allows for close bi-directional communication between the neuron and it surrounding glial coat. Morphological and molecular changes in SGC have been observed in pathological conditions such as inflammation, chemotherapy-induced neuropathy, viral infection and nerve injuries. SGC play critical roles in neuronal excitability and nociception, as well as contribute to promote axon regeneration. Whether findings made in rodent model systems are relevant to human physiology has not been investigated. Here we present a detailed characterization of SGC across species. We characterized the transcriptional profile of SGC in mouse, rat and human at the single cell level. Our findings suggest that key features of SGC in rodent models are conserved in human. These results support the notion that SGC in the sensory system are largely conserved between species. Our study provides the potential to leverage on rodent and human SGC properties and unravel novel mechanisms and potential targets for treating nerve injuries.
Project description:Carrageenan injection induces tissue inflammation and high mechanosensitivity. Dorsal root ganglions contain the cellular body of the sensory neurons which project their axon to the periphery, monitoring this inflammation and transmitting the mechanical stimuli to the spinal cord. To explore whether the sensory neurons underwent transcriptional changes and how long these changes lasted, we injected carrageenan into the left hind paw of mice and sequenced the dorsal root ganglions related to this area 1, 3 and 30 days after the injection.
Project description:We report here a systematic approach to characterize the transcriptional responses of different cells types in the dorsal root ganglion (DRG) to peripheral nerve injury using single cell RNA sequencing (scRNAseq). We compare scRNAseq datasets of lumbar DRGs form naïve mice with corresponding datasets from mice subjected to spared nerve injury (SNI) 7 or 14 days prior to analysis. SNI surgery was performed in the left and right hindleg and development of mechanical allodynia was monitored by von Frey testing relative to control mice and the baseline level. Mice were perfused with PBS prior to extraction of L3 and L4 DRGs and DRGs were enzymatically and mechanically dissociated to single cell suspension before scRNAseq. Analysis of transcriptional changes in this nerve injury-paradigm reveals a differential response at 7 days versus 14 days post injury, suggesting dynamic gene modulation over time.
Project description:Single nucleotide polymorphisms (SNP) can affect mRNA gene expression, in a tissue-specific manner. In this work we survey association of SNP alleles with mRNA gene expression in human dorsal root ganglions (DRG) to gain insights into pathophysiology of pain phenotypes.
Project description:Single nucleotide polymorphisms (SNP) can affect mRNA gene expression, in a tissue-specific manner. In this work we survey association of SNP alleles with mRNA gene expression in human dorsal root ganglions (DRG) to gain insights into pathophysiology of pain phenotypes.
Project description:After an injury in the adult mammalian central nervous system, lesioned axons fail to regenerate. This failure to regenerate contrasts with the remarkable potential of axons to grow following an injury in the peripheral nervous system. Peripheral sensory neurons with cell soma in dorsal root ganglia (DRG) switch to a regenerative state after nerve injury to enable axon regeneration and functional recovery. Decades of research have focused on the signaling pathways elicited by injury in sensory neurons and in Schwann cells that insulate axons as central mechanisms regulating nerve repair. However, neuronal microenvironment is far more complex and is composed of multiple cell types including endothelial, immune and glial cells. Whether the microenvironment surrounding neuronal soma contribute to the poor regenerative outcomes following central injuries remains largely unexplored. To answer this question, we performed a single cell transcriptional profiling of the DRG neuronal microenvironment response to peripheral and central injuries. In dissecting the roles of the microenvironment contribution, we have focused on a poorly studied glia population of Satellite Glial Cells (SGC) surrounding the neuronal cell soma. Upon a peripheral injury, SGC contribute to axon regeneration via Fatty acid synthase (Fasn)-PPARα signaling pathway. Our analysis reveals that in response to central injuries, SGC do not activate the PPAR signaling pathway. However, induction of this pathway with fenofibrate, an FDA- approved PPARα agonist used for dyslipidemia treatment, rescued axon regeneration following an injury to the central nerves. Collectively, our results uncovered a previously unappreciated role of the neuronal microenvironment differential response in central and peripheral injuries.