Project description:Background: Poultry eggs are a low cost, high protein nutrient package that can be consumed as part of a quality diet. However, consumption of poultry egg products has been historically contentious, which highlights the importance of investigating impacts of long-term egg consumption on metabolic health. Objective: Our study utilized the zebrafish Danio rerio, a newly-defined model of human metabolic health, to understand the metabolic consequence of consuming egg products in lieu of other well-described protein sources. Methods: Reference diets were formulated to contain multi-source protein with casein and fish protein hydrolysate (CON; control protein sources), the protein sources of which have been vetted in numerous reference diets. These proteins were then replaced in part with either whole egg (WE; protein and lipid source), egg white (EW; protein source), wheat gluten (WG; cereal protein source), or a high lipid content diet containing a multi-source protein with casein and fish protein hydrolysate (HFCON; isonitrogenous and isolipidic with the WE diet) in a 34-week trial. Daily feeding was initiated at early juvenile life stage and terminated at late reproductive adult stage. Results: The amino acid composition of control vs egg product diets did not vary substantially, although methionine and lysine were apparently limiting in fish fed WG. At termination, fish fed EW as the protein source had similar weight gain and body composition to those fed the CON diet. Blood glucose, both fasting and postprandial, did not differ among any dietary treatment. Assessment of the liver transcriptome using RNAseq revealed no differential gene expression between zebrafish fed CON or WE diets. Zebrafish fed WG had lower weight gain in males. Conclusions: Long-term consumption of egg products promoted metabolic health equal to that of historically-relevant proteins. These data support the value of egg products for maintaining long-term metabolic health in animal diets.
Project description:Egg quality is of pivotal importance in biomedicine, agriculture, ecology and environmental science because of its tremendous influence on reproductive success or failure in all animals. Despite species specific differences in physiological aspects of early development, the evolutionary conserved stereotypical procedure of cellular events, led us to investigate whether these findings are common with marine fish species of aquaculture interest. Atlantic halibut (Hippoglossus hippoglossus) is a highly prized species in global fish markets with increasing demand to its production. The objectives of this study were 1) to reveal the proteomic profiles of good versus poor quality halibut eggs, 2) to identify proteins that can serve as egg quality markers, and 3) to discover molecular mechanisms determining egg quality using a combination of quantitative proteomics workflows, tandem mass tags (TMT) labeling and parallel reaction monitoring (PRM).
Project description:After the end of the last ice age, ancestrally marine threespine stickleback fish (Gasterosteus aculeatus) have undergone an adaptive radiation into freshwater environments throughout the Northern Hemisphere, creating an excellent model system for studying molecular adaptation and speciation. Stickleback populations are reproductively isolated to varying degrees, despite the fact that they can be crossed in the lab to produce viable offspring. Ecological and behavioral factors have been suggested to underlie incipient stickleback speciation. However, reproductive proteins represent a previously unexplored driver of speciation. As mediators of gamete recognition during fertilization, reproductive proteins both create and maintain species boundaries. Gamete recognition proteins are also frequently found to be rapidly evolving, and their divergence may culminate in reproductive isolation and ultimately speciation. As an initial investigation into the contribution of reproductive proteins to stickleback reproductive isolation, we characterized the egg coat proteome of threespine stickleback eggs. In agreement with other teleosts, we find that stickleback egg coats are comprised of homologs to the zona pellucida (ZP) proteins ZP1 and ZP3. We explore aspects of stickleback ZP protein biology, including glycosylation, disulfide bonding, and sites of synthesis, and find many substantial differences compared to their mammalian homologs. Furthermore, molecular evolutionary analyses indicate that ZP3, but not ZP1, has experienced positive Darwinian selection across teleost fish. Taken together, these changes to stickleback ZP protein architecture suggest that the egg coats of stickleback fish, and perhaps fish more generally, have evolved to fulfill a more protective functional role than their mammalian counterparts.
Project description:After the end of the last ice age, ancestrally marine threespine stickleback fish (Gasterosteus aculeatus) have undergone an adaptive radiation into freshwater environments throughout the Northern Hemisphere, creating an excellent model system for studying molecular adaptation and speciation. Stickleback populations are reproductively isolated to varying degrees, despite the fact that they can be crossed in the lab to produce viable offspring. Ecological and behavioral factors have been suggested to underlie incipient stickleback speciation. However, reproductive proteins represent a previously unexplored driver of speciation. As mediators of gamete recognition during fertilization, reproductive proteins both create and maintain species boundaries. Gamete recognition proteins are also frequently found to be rapidly evolving, and their divergence may culminate in reproductive isolation and ultimately speciation. As an initial investigation into the contribution of reproductive proteins to stickleback reproductive isolation, we characterized the egg coat proteome of threespine stickleback eggs. In agreement with other teleosts, we find that stickleback egg coats are comprised of homologs to the zona pellucida (ZP) proteins ZP1 and ZP3. We explore aspects of stickleback ZP protein biology, including glycosylation, disulfide bonding, and sites of synthesis, and find many substantial differences compared to their mammalian homologs. Furthermore, molecular evolutionary analyses indicate that ZP3, but not ZP1, has experienced positive Darwinian selection across teleost fish. Taken together, these changes to stickleback ZP protein architecture suggest that the egg coats of stickleback fish, and perhaps fish more generally, have evolved to fulfill a more protective functional role than their mammalian counterparts.
Project description:Gamete quality is one of the most important element of successful aquaculture. This especially applies to a newly domesticated fish species, such as pikeperch (Sander lucioperca) in which lowered and/or variable egg quality is one of the biggest obstacles toward rapid production expansion. However, mechanisms underlying pikeperch egg developmental competences remain unknown. Therefore, a quantitative proteome study was performed with high (HQ) and low (LQ) quality domesticated pikeperch eggs using a label free LC-MS/MS approach.
Project description:Eggless/SETDB1 (Egg), the only essential histone methyltransferase (HMT) in Drosophila, plays a role in gene repression, including piRNA‐mediated transposon silencing in the ovaries. Previous studies suggested that Egg is post‐translationally modified and showed that Windei (Wde) regulates Egg nuclear localization through protein–protein interaction. Monoubiquitination of mammalian SETDB1 is necessary for the HMT activity. Here, using cultured ovarian somatic cells, we show that Egg is monoubiquitinated and phosphorylated but that only monoubiquitination is required for piRNA‐mediated transposon repression. Egg monoubiquitination occurs in the nucleus. Egg has its own nuclear localization signal, and the nuclear import of Egg is Wde‐independent. Wde recruits Egg to the chromatin at target gene silencing loci, but their interaction is monoubiquitin‐independent. The abundance of nuclear Egg is governed by that of nuclear Wde. These results illuminate essential roles of nuclear monoubiquitination of Egg and the role of Wde in piRNA‐mediated transposon repression.
Project description:In our study, differential male nucleus events and development behaviors were revealed from the fertilized eggs in response to the sperm from males of genotypic sex determination (GSD) and temperature-dependent sex determination (TSD) in gibel carp. When the eggs of maternal fish were fertilized by the sperm from males of GSD, the fertilized egg encountered similar sexual reproduction events and behaviors. However, when the eggs of maternal fish were fertilized by the sperm from males of TSD, a typical process of gynogenesis was observed. To reveal the underlying molecular mechanism of differential sperm nucleus development behaviors in the fertilized eggs, iTRAQ-based quantitative semen proteomics were performed on three semen samples from three males of GSD and three semen samples from three males of TSD respectively.
Project description:The present study is the first study to identify the involvement of circRNAs in the ovary activation and oviposition regulation processes in honey-bee queens.CircRNAs expresion profiles were examined in ovaries of virgin queens, egg-laying queens, egg-laying inhibited queens and egg-laying recovery queens.
Project description:During early stages of development, juvenile fish must rely on their innate immune system to defend against pathogens. At these early stages, the immune system is immature and is unlikely to express the full repertoire of genes that control defences. Although vulnerable, these larval fish can still fight of infections, indicating there are active defence mechanisms. Using rainbow trout as a model, we have taken a transcriptomics approach to determine the antibacterial (Aeromonas salmonicida) and antiviral (VHSV) responses to infection at four early life history stages, eyed egg, post-hatch, first feeding and three weeks post-first feeding. We performed microarray analysis using an Agilent 4x44K custom designed array, using a common RNA reference hybridization design. We show that all stages of the developing fish respond to the disease challenge at 3 days post-challenge, but the number and complexity of the response increases with developmental stage. Specifically, the response to virus at eyed egg and hatch stages does not show the full interferon response as is found at first feeding and 3 weeks post-first feeding. The experiments were carried out with a custom-designed Agilent (Agilent design ID: 028918) oligonucleotide microarray (A-MEXP-2315) named Trout_imm_v1. In total, 46 hybridisations were performed on individual animals, with 4 biological replicates per sampling point (apart from Bacteria infected eyed eggs for which two hybridizations were not used). This work was funded by the European Community’s seventh framework programme (FP7/2007-2013), under grant agreement no. 222719 (LIFECYCLE).
Project description:Somatic cell reprogramming in vitro prior to nuclear transfer is one strategy expected to improve clone survival during development. In this study, we investigated the reprogramming extent of fish fin somatic cells after in vitro exposure to Xenopus egg extract and subsequent culture. Using a cDNA microarray approach, we observed drastic changes in the gene expression profile of the treated cells. Several actors of the TGFbeta and Wnt/beta-catenin signaling pathways, as well as some mesenchymal markers, were inhibited in treated cells, while several epithelial markers were upregulated. This was associated with morphological changes of the cells in culture, suggesting that egg extract drove somatic cells towards a mesenchymal-epithelial transition (MET), the hallmark of somatic reprogramming in iPSCs. However, treated cells were also characterized by a strong decrease in de novo lipid biosynthesis metabolism, the lack of re-expression of pou2 and nanog pluripotency markers, and absence of DNA methylation remodeling of their promoter region. In all, this study showed that Xenopus egg extract treatment initiated an in vitro reprogramming of fin somatic cells in culture. Although not thorough, the induced changes have primed the somatic chromatin for a better embryonic reprogramming upon nuclear transfer.