Project description:The aim of the present study is to summarize the identification of neuropeptides, neuropeptide-like and protein hormones in Z. atratus
Project description:One of the most thoroughly studied insect species, with respect to locomotion behaviour, is the stick insect Carausius morosus. Although detailed information exists on premotor networks controlling walking, surprisingly little is known about neuropeptides, which are certainly involved in motor activity generation and modulation. So far, only few neuropeptides were identified from C. morosus or related stick insects. We performed a transcriptome analysis of the central nervous system to assemble and identify 65 neuropeptide and protein hormone precursors of C. morosus, including five novel putative neuropeptide precursors without clear homology to known neuropeptide precursors of other insects (Carausius neuropeptide-like precursor 1, HanSolin, PK-like1, PK-like2, RFLamide). Using Q Exactive Orbitrap and MALDI-TOF mass spectrometry, 277 peptides including 153 likely bioactive mature neuropeptides were confirmed. Peptidomics yielded a complete coverage for many of the neuropeptide propeptides and confirmed a surprisingly high number of heterozygous sequences. Few neuropeptide precursors commonly occurring in insects, including those of insect kinins and sulfakinins, could neither be found in the transcriptome data nor did peptidomics support their presence. The results of our study represent one of the most comprehensive peptidomic analyses on insects and provide the necessary input for subsequent experiments revealing neuropeptide function in greater detail.
Project description:In a number of species, individuals exposed to pathogens can mount an immune response and transmit this immunological experience to their offspring, thereby protecting them against persistent threats. Such vertical transfer of immunity, named trans-generational immune priming (TGIP), has been described in both vertebrates and invertebrates. Although increasingly studied during the last decade, the mechanisms underlying TGIP in invertebrates are still elusive, especially those protecting the earliest offspring life stage, i.e. the embryo developing in the egg. In the present study, we took the mealworm beetle Tenebrio molitor as a biological model, and combined different proteomic and transcriptomic approaches to determine whether mothers transfer a “signal” (such as fragments of infecting bacteria), mRNA and/or protein/peptide effectors to protect their eggs against two natural bacterial pathogens, namely the Gram-positive Bacillus thuringiensis (Bt) and the Gram-negative Serratia entomophila (Se). We demonstrated that eggs are mainly protected by an active direct transfer of a restricted number of immune proteins and of antimicrobial peptides. In contrast, the present data clearly reject the involvement of mRNA transfer while the transmission of a “signal”, if it happens, is marginal and only occurs within 24h after maternal exposure to bacteria. This work exemplifies the power of combining global multidisciplinary approaches to disentangle the different scenarios of a complex trait, providing a comprehensive characterization of TGIP mechanisms in T. molitor. It also paves the way for future alike studies focusing on TGIP in a wide range of invertebrates and vertebrates to identify additional candidates that could be specific to TGIP and to investigate whether the TGIP mechanisms found herein are specific or common to all insect species.
Project description:RORγt is a transcription factor required for T helper 17 (Th17) cell development. We identified three RORγt-specific inhibitors that suppress Th17 cell responses including Th17 cell-mediated autoimmune disease. We systemically characterized RORγt binding data in the presence and absence of drug with corresponding whole-transcriptome sequencing for wild-type and RORγt-deficient cells. RORγt is central in a densely interconnected regulatory network, acting both as a direct activator of genes important for Th17 cell differentiation and as a direct repressor of genes from other T-cell lineages. The three inhibitors identified here reversed both of these modes of action, but to varying extents and through distinct mechanisms. Whereas one inhibitor displaced RORγt from its target-loci, the two more potent inhibitors affected transcription predominantly without removing DNA-binding. Our work illustrates the power of a system-scale analysis of transcriptional regulation to characterize potential therapeutic compounds that inhibit pathogenic Th17 cells and suppress autoimmunity. DNA binding of RORγt in WT Th17 cells and under chemical perturbations of RORγt; Additional data is included for epitope-tagged exogenous RORγt in EL4 cells (a murine lymphoma cell line)
Project description:The human iPSC line H19101 was differentiated in vitro into cardiomyocytes using a 20-day differentiation protocol (Burridge et al. 2014 PMID 24930130 and Montefiori et al 2018 PMID 29988018 ). 50,000 cardiomyocytes were used in each ATAC-seq experiment. 8 replicates were pooled to obtain the final peak file.
Project description:This study aimed at integrating metabolomics and proteomics data for a comprehensive view of the molecular targets of intervention of protein extracts from Tenebrio molitor in treating hypertension. Serum samples from spontaneously hypertensive rats and Wistar Kyoto rats were analyzed using a quantitative metabolomics and label-free proteomics approach based on liquid chromatography coupled with high resolution mass spectrometry (LC-HRMS). Among deregulated metabolites and proteins in hypertensive rats, we found 15 metabolites and 17 proteins that were restored by supplementation with Tenebrio molitor protein extract. The combination of metabolomics and proteomics provided useful data to uncover the molecular targets of intervention and the underlying functional mechanism of Tenebrio molitor protein extract in an animal model such as spontaneously hypertensive rats. The results suggested that Tenebrio molitor supplementation could effectively treat hypertension, partially by regulating proteins and molecules mainly involved in biological pathways associated to angiotensinogen-angiotensin, Serin protease inhibitors, kallikrein–kinin, reactive oxygen scavenging, and lipid peroxidation.
Project description:Plants from the Nepenthes genus, which enumerates approximately 120 species, possess specialized pitchers enabling them to capture and digest various preys, mainly arthropods, from which the plants derive nutrients. The pitcher fluid contains many molecules of noteworthy importance, including antimicrobial compounds, traditionally used in medicine, as well as hydrolytic enzymes for prey digestion. In this study, polyesters films made from poly(ethylene terephthalate) (PET) and from poly(butylene adipate -co- terephthalate) (PBAT) were incubated in the pitcher of the carnivorous plants Nepenthes alata and Sarracenia purpurea. High performance liquid chromatography analysis revealed hydrolysis into their corresponding monomers, while hydrolysis efficiency was up to ten times higher in the presence of dried mealworm Tenebrio molitor and jasmonic acid. Proteomic analysis indicated the presence of the aspartic proteinase nepenthesin in most conditions, with high abundance in 70% of polyester-containing conditions compared to those without polyesters. Molecular docking simulations further suggested that nepenthesin has the potential to hydrolyze polyesters. While in contrast to cutinases there is only little information on hydrolysis of PET and PBAT by proteases in the literature, this work clearly demonstrates hydrolysis of both PET and PBAT by the recombinant protease nepenthesin, releasing similar amounts of TPA as the cutinase from Humicola insolens. These results suggest carnivorous plant fluids as a source for new enzymes for industrial applications such as for polyester hydrolysis.
Project description:The Crown-of-Thorns starfish (COTS), Acanthaster planci, is a highly fecund predator of reef-building corals distributed throughout the Indo-Pacific. COTS population outbreaks cause substantial loss of coral cover, diminishing the integrity and resilience of the reef ecosystems thus increasing their susceptibility to climate change. We sequenced genomes of A. planci from the Great Barrier Reef, Australia (GBR) and Okinawa, Japan (OKI) to guide identification of species-specific peptide communication with potential applications in mitigation strategies. The genome-encoded proteins excreted and secreted into the surrounding seawater by COTS forming aggregations and by those escaping the predatory giant triton snail, Charonia tritonis, were identified LC-MS/MS.
Project description:Liver are frequently declined for transplantation due to the donor age. It is still unclear how donor age affects the graft quality. Normothermic machine perfusion (NMP) has been proposed as a useful assessment tool prior to transplantation. We aimed to compare the performance of young and elderly rat liver grafts in a small animal NMP model. Grafts from 24 rats were procured, either 3 or 12 months old and perfused for 6 hours at 37°C or stored on ice as a reference group (n=6/group). Livers in both NMP groups cleared lactate and produced bile, with similar pressure, bile production, and pH levels. However, older rat livers showed higher peak transaminase and urea levels. Proteomic analysis revealed differences in protein composition, particularly higher levels of proteins related to oxidoreductase and catalytic activity in older livers. Older age appeared to increase susceptibility to oxidative stress in the livers.
Project description:We aimed to address the challenge of accommodating an increasing demand for aged laboratory animals in the context of rising elderly population and research on aging-related diseases. We developed a cost-effective environmental enrichment method and assessed its impact on metabolic changes in Sprague Dawley rats' livers as a proof-of-concept organ. Twenty-four male rats were divided into four groups, with two kept in standard cages and two in modified rabbit cages. Half of each type of cage group received additional enrichment through weekly playtime in a larger cage. Over six months, the rats' weight gain was consistent across all groups, and corticosterone levels did not significantly differ. However, the control group had significantly lower DHEA and Testosterone levels at the study's end. Rats in enriched environments exhibited less distress during inspections and were more resistant to accepting treats. Animals accustomed to playpen time left their cages more easily. Overall, the study demonstrated that refining husbandry for aging rats is both simple and cost-effective, without detrimental effects on stress levels, development, or liver metabolism.