ABSTRACT: Peptidomics data in support of the study "Detection and characterization of proteolytic enzyme activity and neuropeptide processing in bovine dense core secretory vesicles"
Project description:Proteomics data in support of the study "Detection and characterization of proteolytic enzyme activity and neuropeptide processing in bovine dense core secretory vesicles".
Project description:LC-MS data of MSP-MS assays of CG proteases in support of the study "Detection and characterization of proteolytic enzyme activity and neuropeptide processing in bovine dense core secretory vesicles"
Project description:Neuroendocrine (NE) cells use large dense core vesicles (LDCVs) to traffic, process, store and secrete neuropeptide hormones through the regulated secretory pathway. The DIMM basic helix-loop-helix transcription factor of Drosophila controls the level of regulated secretory activity in NE cells. To pursue its mechanisms, we have performed two independent genome-wide analyses of DIMMM-bM-^@M-^Ys activities: (i) in vivo chromatin immunoprecipitation (ChIP) to define genomic sites of DIMM occupancy and (ii) deep sequencing of purified DIMM neurons to characterize their transcriptional profile. By this combined approach, we showed that DIMM binds to conserved E-boxes in enhancers of 212 genes whose expression is enriched in DIMM-expressing NE cells. DIMM binds preferentially to certain E-boxes within first introns of specific gene isoforms. Statistical machine learning revealed that flanking regions of putative DIMM binding sites contribute to its DNA binding specificity. DIMMM-bM-^@M-^Ys transcriptional repertoire features at least 20 LDCV constituents. In addition, DIMM notably targets the pro-secretory transcription factor, CREB-A, but significantly, DIMM does not target any neuropeptide genes. DIMM therefore prescribes the scale of secretory activity in NE neurons, by a systematic control of the regulated secretory pathway at steps that are both proximal and distal. DIMM::MYC ChIP-chip (c929>DIMM::MYC/tubGAL80ts) and control (c929>tubGAL80ts): 2 replicates each, input and IP samples. Total of 8 arrays
Project description:Neuroendocrine (NE) cells use large dense core vesicles (LDCVs) to traffic, process, store and secrete neuropeptide hormones through the regulated secretory pathway. The DIMM basic helix-loop-helix transcription factor of Drosophila controls the level of regulated secretory activity in NE cells. To pursue its mechanisms, we have performed two independent genome-wide analyses of DIMM’s activities: (i) in vivo chromatin immunoprecipitation (ChIP) to define genomic sites of DIMM occupancy and (ii) deep sequencing of purified DIMM neurons to characterize their transcriptional profile. By this combined approach, we showed that DIMM binds to conserved E-boxes in enhancers of 212 genes whose expression is enriched in DIMM-expressing NE cells. DIMM binds preferentially to certain E-boxes within first introns of specific gene isoforms. Statistical machine learning revealed that flanking regions of putative DIMM binding sites contribute to its DNA binding specificity. DIMM’s transcriptional repertoire features at least 20 LDCV constituents. In addition, DIMM notably targets the pro-secretory transcription factor, CREB-A, but significantly, DIMM does not target any neuropeptide genes. DIMM therefore prescribes the scale of secretory activity in NE neurons, by a systematic control of the regulated secretory pathway at steps that are both proximal and distal.
Project description:The goal of this experiment was to determine the RNA contents of extracellular vesicles isolated from 3-6mm bovine ovarian follicles.
Project description:Phosphopeptidomic and peptidomic profiling of dense core secretory vesicles. Phosphosite occupancy was measured by comparing non-phosphorylated peptide intensity before and after treatment by alkaline phosphatase,
Project description:As a model hemimetabolous insect species and an invasive urban pest that is globally distributed, the American cockroach, Periplaneta americana, is of great interest in both basic and applied research. Previous studies on P. americana neuropeptide identification have been based on biochemical isolation and molecular cloning. In the present study, an integrated approach of genomics- and peptidomics-based discovery was performed for neuropeptide identification in this insect species. Using large-scale peptidomic analysis of peptide extracts from 4 different tissues (the central nervous system, corpora cardiac and corpora allata complex, midgut, and male accessory gland), 35 conserved (predicted) and a potential (novel) neuropeptides were then identified. Subsequent experiments revealed the tissue distribution, sex difference, and developmental patterns of 2 conserved neuropeptides (allatostatin B and short neuropeptide F) and a novel neuropeptide PaOGS36577. Our study shows a comprehensive neuropeptidome and detailed spatiotemporal distribution patterns, providing a solid basis for future functional studies of neuropeptides in the American cockroach.
Project description:In order to gain further insight into the molecular mechanism(s) mediating the blunted epinephrine responses following recurrent hypoglycemia we utilized global gene expression profiling approach. Our results indicate the association between defective counterregulation (impaired epinephrine release) and the activation of the unfolded protein response as well as increased neuropeptide signaling, altered ion homeostasis and downregulation of proteins involved in Ca2+-dependent exocytosis of secretory vesicles. We compared the entire transcriptomes during recurrent (defective CRR model, 2RH) and acute hypoglycemia (normal CRR group, 2RS) in the adrenal medulla of normal Sprague-Dawley rats.
Project description:In order to gain further insight into the molecular mechanism(s) mediating the blunted epinephrine responses following recurrent hypoglycemia we utilized global gene expression profiling approach. Our results indicate the association between defective counterregulation (impaired epinephrine release) and the activation of the unfolded protein response as well as increased neuropeptide signaling, altered ion homeostasis and downregulation of proteins involved in Ca2+-dependent exocytosis of secretory vesicles.
Project description:Objectives: MicroRNA (miRNA) can be released to the extracellular medium and participates in neuronal communication. We investigate the mechanisms of miRNA exocytosis by vesicle fusion as a neuromodulator in a manner that are disparate from silencing gene expression. Methods: Small RNA sequencing data of large dense-core vesicle were generated by next-generation sequencing (NGS) in triplicate using Illumina Hiseq 2500. Results: Large dense-core vesicles contain a variety of known and novel miRNAs inside including miR-375. Conclusion: miRNAs can be novel neuromodulators, which are stored in LDCVs and released by vesicle fusion by SNARE assembly and synaptotagmin-1