Project description:We directed mESCs towards the dorsal sensory interneurons (dIs) using RA-/+ BMP4 and tested the effects of an axon guidance molecule netrin1 on dI differentiation in vitro. Using bulk-RNA seq analysis, we investiagted when the addition of netrin 1 will affect the transcriptome of differentiating dIs. For this analysis, we used three timewindows (conditions 1-3) for netrin addition (1) condition 1 netrin 1 was added together with RA+BMP4 at day 4, and RNA seq was conducted at day 5, (2) netrin1 was added after the RA+BMP4 mediated patterning at day 5 and RNA seq was conducted at day 6, and (3) netrin1 was added for the extended duration between day 5-day9 to investigate the netrin1 effects throughout the dI differentiation and RNA seq was conducted at day 9. Through this analysis we identified that netrin1 downregulate BMP signaling pathway, possibly through modlulating the mRNA processing of key mediator of BMP signaling such as Id genes and BMP signaling inhibitor Smad7 and affect dI differentiation. This is the first report to show that netrin1 regulate dI fate specification in the dorsal spinal cord.
Project description:This experiment aims at characterizing the transcriptome of embryonic mouse dorsal spinal cord. Dorsal spinal cords dissected from litters of E14.5 wild type embryos of unknown sex were processed for RNA extraction using Trizol and RNeasy Mini kit (Qiagen) extraction procedures. Five replicates of wild type embryos were analyzed, each sample with tissue pooled from three embryos.
Project description:Here we performed a ChIP-seq experiment for Tlx3 trancription factor on a sample of mouse embryonic dorsal spinal cord. The result is the generation of the genome-wide maps for Tlx3 binding to chromatin in dILB neurones of the developing dorsal horn.
Project description:To identify differentially expressed genes in the developmental mouse dorsal spinal cord, we characterized the global gene expression profiling of mouse embryonic dorsal spinal cord commissural neurons at E10.5, E11.5 and E12.5. We used the Affymetrix Mouse Exon 1.0 ST Array platform to analyze the gene expression profiling. We included the gene expression data obtained from dorsal spinal cord commissural neuron at different embryonic stage. 2 Biological replicates were performed.
Project description:Our study examined a population of radial glial-like cells (RGLCs) in the dorsal spinal cord midline that we showed provide long-distance growth support for longitudinal rapidly adapting (RA) mechanoreceptor axons during development of spinal cord dorsal column. To evaluate potential molecular markers of these cells, we isolated the RGLCs using hematoxylin and eosin staining to visualize the cell bodies in the dorsal column midline from E14.5 mouse embryos, and used laser capture microdissection for each sample ("LCM"). To compare transcript expression to the adjacent RA mechanoreceptive axons, we performed FACS of dorsal root ganglion of E14.5 Ret-Tdtomato+ RA mechanoreceptors. Our analyses revealed a high enrichment of radial glial-specific markers in the LCM replicates compared to Ret-Tdt samples. In contrast, neuronal-specific markers were more highly enriched in the Ret-Tdt samples, as expected. These data suggest the midline RGLCs are of radial glial identity. Others may find these data helpful in determining potential RGLC-mechanoreceptor molecular interactions in subsequent studies.
Project description:To study the gene expression at the level of dorsal root ganglia and spinal cord in acute herpetic neuralgia mice, we induced acute herpetic neuralgia by subcutaneous injection of HSV-1 virus fluid into the tibia of C57/6J mice. We collected data obtained from RNA-seq of dorsal root ganglia and spinal cord tissue in acute herpetic neuralgia mice (7 days after virus injection) and control mice for gene expression profiling.
Project description:Spinal cord injury (SCI) is a devastating clinical condition resulting in significant disabilities for affected individuals. Apart from local injury within the spinal cord, SCI patients develop a myriad of complications characterized by multi-organ dysfunction. Some of the dysfunctions are directly related to the disrupted integrity of sensory afferents from DRGs, which signal to both the spinal cord and peripheral organs. Some classes of DRG neurons undergo axonal sprouting both peripherally and centrally after spinal cord injury. Such physiological and anatomical re-organization of afferent axons after SCI contributes to both adaptive and maladaptive plasticity, which may be modulated by activity/exercise. In this study, we collected comprehensive gene expression data in whole dorsal root ganglia (DRGs) throughout the levels below the injury comparing the effects of SCI with and without activity/exercise.
Project description:Genes are up and down regualted in DRG and spinal dorsal cord after peripheral nerve injury WT male adult with sciatic and femoral nerve transection 7 days, RNA was purified from ipilateral or contralateral L4-L6 DRGs or lumbar spinal dorsal cords
Project description:Peripheral somatosensory input is modulated in the dorsal spinal cord by a network of excitatory and inhibitory interneurons. PTF1A is a transcription factor essential in dorsal neural progenitors for specification of these inhibitory neurons. Thus, mechanisms regulating Ptf1a expression are key for generating neuronal circuits underlying somatosensory behaviors. Mutations targeted to distinct cis-regulatory elements for Ptf1a in mice, tested the in vivo contribution of each element individually and in combination. Mutations in an auto-regulatory enhancer resulted in reduced levels of PTF1A, and reduced numbers of specific dorsal spinal cord inhibitory neurons, particularly those expressing Pdyn and Gal.