To explore the molecular basis of the distinct intrinsic membrane properties and other dstinguishing features of functionally defined DRG neuron subtypes, we bulk-sequenced RNA at high depth of genetically-labeled DRG neurons to generate transcriptome profiles of eight major DRG neuron subtypes. The trancriptome profiles revealed differentially expressed and functionally relevant genes, including voltage-gated ion channels. Guided by the transcriptome pofiles, electrophysiological analyses using pharmacological and genetic manipulations as well as computational modeling of DRG neuron subtypes were undertaken to assess the functions of select voltage-gated potassium channels (Kv1, Kv2, Kv3, and Kv4) in shaping action potential (AP) waveforms and firing patterns of the DRG neuron subtypes. Our findings show that the transcriptome profiles have predictive value for defining ion channel contributions to sensory neuron subtype-specific intrinsic physiological properties. Overall design: We made use of genetic tools developed that selectively label each of eight DRG neuron subtypes with fluorescent reporters, first purifying labeled neurons to homogeneity using flow cytometry (FACS) and then extracting RNA from these purified neuronal populations. DRGs from all axial levels were used for FACS, except for proprioceptors which were purified from thoracic ganglia, because the PVIRES-Cre; Ai14 proprioceptor labeling strategy was found to label a subset of limb level cutaneous LTMRs as well. DRGs from multiple mice were combined for FACS, and neurons from multiple rounds of sorting were combined to obtain sufficient amounts of RNA for each sequencing reaction. At least three biological replicates with a total of 26 samples were sequenced for each neuronal subtype. RNA libraries were prepared and subsequently sequenced using an Illumina HiSeq2000 platform at an average depth of ~70 million mapped reads per sample. This depth translates to an average detection level of 12,226 genes per sample. By using well characterized mouse lines for specific labeling of neuronal subtypes, this analysis links gene expression patterns to sensory neuron subtypes defined by their distinct in vivo properties as well as their corresponding intrinsic properties.
Deep Sequencing of Somatosensory Neurons Reveals Molecular Determinants of Intrinsic Physiological Properties.
No sample metadata fields
View SamplesThe development of complex tissues requires that mitotic progenitor cells integrate information from the environment. The highly varied outcomes of such integration processes undoubtedly depend at least in part upon variations among the gene expression programs of individual progenitor cells. To date, there has not been a comprehensive examination of these differences among progenitor cells of a particular tissue. Here, we used comprehensive gene expression profiling to define these differences among individual progenitor cells of the vertebrate retina. Retinal progenitor cells (RPCs) have been shown by lineage analysis to be multipotent throughout development and to produce distinct types of daughter cells in a temporal, conserved order. A total of 42 single RPCs were profiled on Affymetrix arrays. An extensive amount of heterogeneity in gene expression among RPCs, even among cells isolated from the same developmental time point, was observed. While many classes of genes displayed heterogeneity of gene expression, the expression of transcription factors constituted a significant amount of the observed heterogeneity. Additionally, the expression of cell cycle related transcripts showed differences among those associated with G2 and M, versus G1 and S phase, suggesting different levels of regulation for these genes. These data provide insights into the types of processes and genes that are fundamental to cell fate choices, proliferation decisions, and, for cells of the central nervous system, the underpinnings of the formation of complex circuitry.
Individual retinal progenitor cells display extensive heterogeneity of gene expression.
Specimen part
View SamplesIdentification of genes expressed in a preferential manner in the developing ciliary body/iris will provide a starting point for future functional analyses. To identify candidate genes expressed in a variety of ocular tissues during development, we have profiled single cells from the developing eye. Post hoc identification of the origin of these cells showed that they included cells from the periphery of the developing optic cup. By comparing the expression profiles of these cells to many retinal cell types, candidate genes for preferential expression in the periphery were identified.
Identification of genes expressed preferentially in the developing peripheral margin of the optic cup.
Specimen part
View SamplesThe vertebrate retina uses diverse neuronal cell types arrayed into complex neural circuits to extract, process and relay information from the visual scene to the higher order processing centers of the brain. Amacrine cells, a diverse class of inhibitory interneurons, are thought to mediate the majority of the processing of the visual signal that occurs within the retina. Despite morphological characterization, the number of known molecular markers of amacrine cell types is still much smaller than the 26 morphological types that have been identified. Furthermore, it is not known how this diversity arises during development. Here, we have combined in vivo genetic labeling and single cell genome-wide expression profiling to: 1) Identify specific molecular types of amacrine cells; 2) Demonstrate the molecular diversity of the amacrine cell class.
Development and diversification of retinal amacrine interneurons at single cell resolution.
No sample metadata fields
View SamplesIn this study, we examine the consequences of the loss of two related factors, Onecut1 and Onecut2, during mouse retinal development.
Onecut1 and Onecut2 play critical roles in the development of the mouse retina.
Specimen part
View SamplesIn this study, we examine the consequences of the loss of two related factors, Onecut1 and Onecut2, during mouse retinal development and maturation.
Onecut1 and Onecut2 play critical roles in the development of the mouse retina.
Specimen part
View SamplesDetailed analysis of genome-wide transcriptome profiling in rice root is reported here, following Cr-plant interaction. Such studies are important for the identification of genes responsible for tolerance, accumulation and defense response in plants with respect to Cr stress. Rice root metabolome analysis was also carried out to relate differential transcriptome data to biological processes affected by Cr (VI) stress in rice.
Transcriptomic and metabolomic shifts in rice roots in response to Cr (VI) stress.
Age, Specimen part, Treatment
View SamplesThis SuperSeries is composed of the SubSeries listed below.
Polo-Like Kinase 3 Appears Dispensable for Normal Retinal Development Despite Robust Embryonic Expression.
Specimen part
View SamplesThese data investigate the transcriptomic differences in the whole retinas of mice resulting from loss of Polo-like Kinase 3 (Plk3) over various stages of development, including adulthood, postnatal day (P)7, and P0.
Polo-Like Kinase 3 Appears Dispensable for Normal Retinal Development Despite Robust Embryonic Expression.
Specimen part
View SamplesAnalysis of adult retinas from tripartite motif-containing domain 9 knockouts and wild type littermates. Trim9 belongs to the TRIM family of E3 ubiquitin ligases. Results provide insight into possible roles for Trim9 in the retina.
The Trim family of genes and the retina: Expression and functional characterization.
Specimen part
View Samples