Proteomics coupled with in vitro model to study the early crosstalk occurring between newly excysted juveniles of Fasciola hepatica and host intestinal cells
Ontology highlight
ABSTRACT:
INSTRUMENT(S): TripleTOF 5600
ORGANISM(S): Fasciola Hepatica (liver Fluke)
SUBMITTER: 
David Becerro Recio
LAB HEAD: Mar Siles-Lucas
PROVIDER: PXD033952 | Pride | 2022-09-20
REPOSITORIES: Pride
Similar Datasets
Project description:The project presents the set-up of an ex vivo model coupled with quantitative SWATH-MS proteomics to study the early events following the passage of Fasciola hepatica Newly Excysted Juveniles (NEJ) though the host’s intestinal wall, assessing the proteomic changes in their surface (tegument) and soma in response to host stimuli.
2022-08-31 | PXD033945 | Pride
Project description:Solid State Fermentation (SSF) processes have been explored for yeast growth and protein and metabolites production. However, most of these processes lack standardization. In this work, we present a polylactic acid (PLA) 3D printed matrix that dramatically enhances yeast growth when embedded in liquid media compared to equivalent static cultures, and changes yeast expression patterns at the proteome level. Moreover, differences in sugar assimilation and ethanol production, as the main product of alcoholic fermentation, are observed. Our results suggest that these matrixes may be useful for a vast range of biotechnological applications based on yeast fermentation.
2023-09-07 | PXD043759 | Pride
Project description:The DNA, its epigenetic information and signals from the direct environment, all work together to orchestrate the development of the single fertilized oocyte into a new individual. The concept of developmental programming, attributed to the embryonic plasticity, suggests that the early life environment influences offspring characteristics in later life, but also further generations. There is increasing awareness about the use of Assisted Reproductive Technologies (ART), because, due to its high inefficiency mimicking in vivo optimal conditions, implies an extraordinary change in the environment where the beginning of a new organism takes place. Here we assessed the transgenerational effects of the cryopreserved embryo replacement on a maternal tract using the rabbit as a model. Then, one progeny derived from vitrified-transferred embryos (VT group) was compared to a cohort of animals naturally-conceived (NC group). A multi-omic approach (metabolome, proteome and epigenome) were addressed on the liver tissue in order to complete or previous phenotypic and transcriptomic characterization of F3 animals. LC-ESI-MS and LC-APCI-MS analysis were performed to interrogate the semi-polar and non-polar metabolomes, respectively. For the proteomic study, the analysis was carried on by LC-SWATH-MS. Finally, genome-wide DNA methylation profiling was studied by MBD-Seq. The healthy state was evaluated by a haematological and biochemical analysis of the peripheral blood. Targeted and untargeted metabolome analysis revealed a global metabolome alteration in the VT livers, particularly for those pathways related with the lipid metabolism. Here was detected some disturbances in the metabolism of long-chain polyunsaturated fatty acids (LC-PUFA), which was corroborated by the proteomic data. Functional analysis of the subset of proteins that differs between VT and NC livers revealed significant changes in the metabolism of the linoleic (LIN) and arachidonic (ARA) acids, whose downstream consequences directly was related with dysregulations in the growth and immune system responses. Overall results denoted that these metabolic disturbances participated in a complex network of physiological pathways that together promotes the phenotypic deviations currently observed after an embryo cryopreservation-transfer procedure (CTP). The underlying hypothesis supported by our results, is that these developmental deviations respond to changes in the epigenome, as consequence of direct perturbation of the reprogramming process by ART, which are dragged until adult life and inherited by further generations. Changes in the epigenetic suggested possible interactions between lipid and apoptotic pathways that jointly reprograms the physiology of the liver. However, based on peripheral blood parameters, the healthy status of VT animals was comparable to that of the NC group. Our data provides valuable information about the transgenerational long-term molecular effects of the embryo CTP using high-throughput sciences, which should highlight the need to study ART not only to increase pregnancy rates, but also to use ART in the safest way possible.
2022-02-15 | PXD017972 | Pride
Project description:Neural stem cells (NSCs) in the adult mammalian subependymal zone maintain a glial identity and the developmental potential to generate neurons during the lifetime. Production of neurons from these NSCs is not direct but follows an orderly pattern of cell progression which allows the gradual increase along the neurogenic lineage in the expression of pro-neural factors needed for neuronal specification. In this context, tightly regulated translation of existing transcriptional programs represents a potential mechanism to avoid the critical challenge posed by genes that encode proteins with conflicting functions, i.e. self-renew or differentiate. Here, we identify RNA-binding protein MEX3A as a post-transcriptional regulator of a set of stemness-associated transcripts at critical transitions in the subependymal neurogenic lineage. MEX3A binding to a set of quiescence-related RNAs in activated NSCs is needed for their return to quiescence, playing a role in the long-term maintenance of the NSC pool. Furthermore, it is required for the repression of the same program at the onset of neuronal differentiation. Our data indicate that MEX3A is a pivotal regulator of adult mammalian neurogenesis acting as a translational remodeller.
2023-01-08 | PXD038983 | Pride
Project description:Common octopus, Octopus vulgaris, is an economically important cephalopod species. However, its rearing under captivity is currently challenged by massive mortalities previous to their juvenile stage due to nutritional and environmental factors. Dissecting the genetic basis and regulatory mechanism behind this mortality requires genomic background knowledge. A transcriptomic sequencing of 10 dph octopus paralarvae from different experimental conditions was constructed via RNA-seq. A total of 613,767,530 raw reads were filtered and de novo assembled into 363,527 contigs of which 82,513 were annotated in UniProt carrying also their GO and KEGG information. Differential gene expression analysis was carried out on paralarvae reared under different diet regimes and temperatures, also including wild paralarvae. Genes related to lipid metabolism exhibited higher transcriptional levels in individuals whose diet includes crustacean zoeas, which had an impact over their development and immune response capability. High temperature induces acclimation processes at the time that increase metabolic demands and oxidative stress. Wild individuals show an expression profile unexpectedly similar to Artemia fed individuals. Proteomic results support the hypothesis revealed by transcriptional analysis. The comparative study of the O. vulgaris transcriptomic profiles allowed the identification of genes that deserve to be further studied as candidates for biomarkers of development and health. The results obtained here on the transcriptional variations of genes caused by diet and temperature will provide new perspectives in understanding the molecular mechanisms behind nutritional and temperature requirements of common octopus that will open new opportunities to deepen in paralarvae rearing requirements.
2019-11-12 | PXD014141 | Pride
Project description:Since the first human conceived through in vitro fertilisation in 1978, over 8 million babies have been born by assisted reproductive technologies (ART). Although most ART babies and children seem healthy, in recent years, several human and animal model studies have evidenced a potential impact of ART on long-term development. However, the long-term follow-up data in this field is still limited. Till now, studies are mainly focused on techniques such as in vitro fertilisation or in vitro culture, being the information from gametes/embryos cryopreservation field practically missing. Herein, we have developed an animal model to determine whether vitrified-thawed embryo transfer procedure has long-term consequences over the offspring. The birth weight, growth performance and adult body weight of the rabbits derived from vitrified-thawed embryos was compared with that of the naturally-conceived animals. In adulthood, the liver, heart, kidneys, spleen, lungs, gonads and adrenal glands of the males were weighed and compared. Moreover, some haematological and biochemical parameters were assessed on peripheral blood. Besides, some liver samples were obtained to perform a comparative proteomic study. The embryo vitrification-transfer process modified the birth weight and the growth pattern of the offspring, reducing the growth performance in a sex-specific manner. In adulthood, animals derived from vitrified embryos showed a significantly lower body, liver and heart weight. Molecular analyses revealed that vitrified-thawed embryo transfer procedure triggers concordant reprogramming of the liver proteome. The most relevant metabolic alteration denoted by the protein profile was that related to the oxidative phosphorylation, suggesting an impaired oxidative metabolism in the mitochondria. Furthermore, hints of dysregulation in the zinc and lipid metabolism were identified. These results evidence long-term consequences in the offspring derived from cryopreserved-transferred embryos and represented an evident example of the phenotypic plasticity exhibited by the mammalian embryo.
2021-09-09 | PXD016874 | Pride
Project description:In order to investigate proteolytic activity (presence of peptidases) in eggs of F. hepatica, we performed LC-MS analysis of eggs. Eggs were colelcted fresh and then after 5 and 10 days of incubation (embryonation). Each time point done in triplicates.
2022-07-11 | PXD022516 | Pride
Project description:Breeding schemes for meat production in rabbits involved a three-way cross of specialized lines in which a paternal line inseminates maternal crossbred females. Paternal line or terminal sire are selected for growth traits, being the males used for the production of fertile dose at the insemination centres and farms. So high growth rate males must produce, in addition, semen in sufficient quantity and quality to meet the demand of insemination and, nevertheless, several studies have been showed that selection for growth have effects on reproduction performance in female and males. In rabbits, negative effects has been observed in ovulation induction, prenatal survival and genetic correlation to fertility. Many factors influence the production and quality of rabbit semen, management as collection frequency, environment (season or photoperiod, nutrition and genetic. Most of the previous studies have been focused in the effects of selection on the seminal and sperm parameters but, little attention has been paid to the protein seminal plasma or sperm composition and if these changes could be affect the fertility of seminal doses obtained from the paternal males. The aim of this study was to evaluate if selection program by daily gain in fattening period has changed seminal traits, plasma and sperm proteoma and, the fertility of semen when it is used in artificial insemination. To do this we uses two re-derived groups of paternal males obtained from vitrified embryos with a difference of 18 generations between both groups.
2020-01-13 | PXD015511 | Pride
Project description:Breeding schemes for meat production in rabbits involved a three-way cross of specialized lines in which a paternal line inseminates maternal crossbred females. Paternal line or terminal sire are selected for growth traits, being the males used for the production of fertile dose at the insemination centres and farms. So high growth rate males must produce, in addition, semen in sufficient quantity and quality to meet the demand of insemination and, nevertheless, several studies have been showed that selection for growth have effects on reproduction performance in female and males. In rabbits, negative effects has been observed in ovulation induction, prenatal survival and genetic correlation to fertility. Many factors influence the production and quality of rabbit semen, management as collection frequency, environment (season or photoperiod, nutrition and genetic. Most of the previous studies have been focused in the effects of selection on the seminal and sperm parameters but, little attention has been paid to the protein seminal plasma or sperm composition and if these changes could be affect the fertility of seminal doses obtained from the paternal males. The aim of this study was to evaluate if selection program by daily gain in fattening period has changed seminal traits, plasma and sperm proteoma and, the fertility of semen when it is used in artificial insemination. To do this we uses two re-derived groups of paternal males obtained from vitrified embryos with a difference of 18 generations between both groups.
2020-01-13 | PXD015510 | Pride
Project description:Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate with the loss of sperm function during cryopreservation remains unsolved. The present study attempted to clarify this issue evaluating differential changes in the proteome of pig spermatozoa retrieved from the cauda epididymis and the ejaculate, with clear differences in cryotolerance, comparing fresh and frozen-thawed cells. Sperm samples were collected from 10 healthy, sexually mature and fertile boars, and cryopreserved using a standard 0.5 mL straw protocol. Total and progressive motility, viability and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) cauda epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT sperm samples were analyzed using a LC-ESI-MS/MS-based SWATH approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins directly involved in mitochondrial functionality among those quantitatively altered in ejaculated spermatozoa, which would explain the worse post-thaw quality of ejaculated pig spermatozoa.
2019-11-11 | PXD012984 | Pride