Project description:We report the application of ribosomal profiling based RNA sequencing technology for high-throughput profiling of the Gr5a cells of male Drosophila melanogaster adults. By expressing the UAS-Rpl3-3XFLAG transgene using the Gr5a-GAL4 driver.
Project description:We report the application of targeted DNA Adenine Methyltransferase identification DNA sequencing technology for high-throughput profiling of Pcl occupancy in the Gr5a cells of male Drosophila melanogaster adults. By expressing the UAS-LT3-Dam::Pcl and UAS-LT3-Dam transgene using the Gr5a-GAL4;tubulinGAL80ts driver.
Project description:We report the application of targeted DNA Adenine Methyltransferase identification DNA sequencing technology for high-throughput profiling of OGT occupancy in the Gr5a cells of male Drosophila melanogaster adults. By expressing the UAS-LT3-Dam::Pcl and UAS-LT3-Dam transgene using the Gr5a-GAL4;tubulinGAL80ts driver.
Project description:Taste stem/progenitor cells from the mouse posterior tongue have been recently used to generate taste bud organoids. However, the inaccessible location of the taste receptor cells is observed in conventional organoids. Here, we established a suspension culture method for fine tuning of taste bud organoid by apicobasal polarity alteration to form the accessible localization of taste receptor cells in organoid. Compared to conventional Matrigel-embedded organoids, suspension-cultured organoids showed comparable differentiation and renewal rates to those of taste buds in vivo and exhibited functional taste receptor cells and cycling progenitor cells. Accessible taste receptor cells on the outer region of taste bud organoids enabled the direct application of calcium imaging for evaluating the taste response. Moreover, suspension-cultured organoids could be genetically altered using gene editing methods. Suspension-cultured taste bud organoid harmoniously integrated with the recipient lingual epithelium; maintained the taste receptor cells and gustatory innervation capacity. Thus, we propose that suspension-cultured organoids may provide efficient model for taste research including taste bud development, regeneration and transplantation
Project description:To uncover novel molecules involved in taste detection, we performed a microarray-based screen for genes enriched in taste neurons. Proboscis RNA from flies homozygous for a recessive poxn null mutation was compared to RNA from heterozygous controls. Poxn mutants have a transformation of labellar gustatory chemosensory bristles into mechanosensory bristles and therefore lack most or all taste neurons. Experiment Overall Design: Proboscises of poxn70 homozygous mutant and poxn70 heterozygous mutant males (8-18 days post eclosure) were dissected, and total RNA was harvested in Trizol according to standard trizol protocol. Samples for each microarray were prepared from 164-280 proboscises. We performed 3 biological replicates for each genotype.
Project description:Purpose: The goal of this study is to compare the taste bud transcriptome of fasted and fed mice. By doing this we hope to gain more information about the taste bud transcriptome as well as determine transcriptional changes that may be linked with pre or post-ingestive states. Methods: Whole taste bud mRNA profiles of WT adult C57BL/6J mice were generated by Illumina HiSeq 2000 single end RNA sequencing. Prior to taste bud RNA isolation, mice were either food-deprived, fed, or given ad libitum access to standard rodent chow as a control. Sequence read quality was assesed using FastQC, and adapter sequences were trimmed using trimmomatic. Reads were then aligned to the GRCm38.p4 reference genome using Tophat2 followed by Cufflinks analysis, then gene level differential expression was conducted using Cuffdiff, and visualized via cummeRbund. Gene enrichment analysis of differentially expressed genes was performed using DAVID and Panther Gene Ontology. Results: We mapped about 30 million sequence reads per sample to the mouse genome, build GRCm38.p4, and identified 144 differentially expressed genes (FDR of 5%) between the taste buds of food-restricted, fed, and ad libitum control mice. Gene enrichment analysis of differentially expressed genes showed enrichment of pathways associated with cytokines, immunity, cytoskeletal structure, chaperone proteins, and protease inhibitors. Conclusions: Our study represents a detailed analysis of the murine taste bud transcriptome generated using RNA-seq technology. Our results highlight cellular pathways that may be differentially regulated in taste receptor cells during different pre/post-ingestive states. Additionally, as limited transcriptome information is available for taste buds, this dataset can serve as a resource for the discovery of genes novel to the taste bud.