RESUMO
The nematode intestine is the primary site for nutrient uptake and storage as well as the synthesis of biomolecules; lysosome-related organelles known as gut granules are important for many of these functions. Aspects of intestine biology are not well understood, including the export of the nutrients it imports and the molecules it synthesizes, as well as the complete functions and protein content of the gut granules. Here, we report a mass spectrometry (MS)-based proteomic analysis of the intestine of the Caenorhabditis elegans and of its gut granules. Overall, we identified approximately 5,000 proteins each in the intestine and the gonad and showed that most of these proteins can be detected in samples extracted from a single worm, suggesting the feasibility of individual-level genetic analysis using proteomes. Comparing proteomes and published transcriptomes of the intestine and the gonad, we identified proteins that appear to be synthesized in the intestine and then transferred to the gonad. To identify gut granule proteins, we compared the proteome of individual intestines deficient in gut granules to the wild type. The identified gut granule proteome includes proteins known to be exclusively localized to the granules and additional putative gut granule proteins. We selected two of these putative gut granule proteins for validation via immunohistochemistry, and our successful confirmation of both suggests that our strategy was effective in identifying the gut granule proteome. Our results demonstrate the practicability of single-tissue MS-based proteomic analysis in small organisms and in its future utility.
Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Lisossomos , Proteômica , Animais , Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteômica/métodos , Lisossomos/metabolismo , Proteoma/metabolismo , Intestinos , Mucosa Intestinal/metabolismo , Gônadas/metabolismo , Espectrometria de Massas/métodos , Organelas/metabolismoRESUMO
BACKGROUND: Gel property is among the crucial functional properties of egg yolk (EY), which determines the texture and flavor of EY products. In the present study, the effects of two unsaturated fatty acids [monounsaturated fatty acid oleic acid (OA) and diunsaturated fatty acid linoleic acid (LA)] on the gel properties of EY protein were investigated. RESULTS: Compared with the blank group, the addition of LA and OA (10-50 g kg-1) improved the gel hardness (from 270.54 g to 385.85 g and 414.38 g, respectively) and viscosity coefficient (from 0.015 Pa.sn to 11.892 Pa.sn and 1.812 Pa.sn, respectively). The surface hydrophobicity of EY protein increased to a maximum value of 40 g kg-1 with the addition of both fatty acids (39.06 µg and 41.58 µg, respectively). However, excess unsaturated fatty acids (≥ 50 g kg-1) disrupted the completeness of the gel matrix and weakened the structural properties of the EY gel. CONCLUSION: Both fatty acids improved the gel properties of EY protein. At the same addition level, OA was superior to LA in improving gel properties. The present study provides a theoretical underpinning for the sensible application of unsaturated fatty acids in improving EY gel properties. © 2024 Society of Chemical Industry.
Assuntos
Galinhas , Proteínas do Ovo , Gema de Ovo , Géis , Interações Hidrofóbicas e Hidrofílicas , Ácido Linoleico , Ácido Oleico , Ácido Linoleico/química , Ácido Oleico/química , Viscosidade , Géis/química , Proteínas do Ovo/química , Gema de Ovo/química , Animais , DurezaRESUMO
Zeugodacus cucurbitae (Coquillett) is an important pest of fruit and vegetable crops in tropical and subtropical regions. Previous studies have shown that short-term high-temperature stress has a significant effect on the oviposition behavior of three successive generations (F1 -F3 ) of Z. cucurbitae (Coquillett). For the clarification of the molecular response of the oviposition behavior of Z. cucurbitae (Coquillett) to short-term high-temperature stress, three Vitellogenin (Vg) genes, namely, Vg-1, Vg-2, and Vg-3, and one Vitellogenin receptor (VgR) gene were selected; 25°C was used as the control treatment; and 33°C, 37°C, 41°C, and 45°C were set as the high-temperature treatments. Newly emerged adults of the F1 generation were treated for 1 h, and the expression dynamics of the target genes were analyzed 7 days after the emergence of three successive generations of adults. Results showed that the expression of the Vg gene in the 33°C and 37°C groups was upregulated compared with that in the control group, and the difference among the 41°C, 45°C, and control groups was small. VgR gene expression level gradually increased in each treatment group with the increase in the number of days and peaked on Days 6 and 7. Compared with the control group, the expression of VgR gene in the F1 generation was downregulated in the high-temperature treatment group over 7 days. On Day 7, the expression level of the VgR gene in the F2 and F3 generations in the 37°C and 45°C groups was significantly higher than that in the F2 and F3 generations in the control group. In conclusion, Vg and VgR are transformed and utilized differently after short-term high-temperature treatment.
Assuntos
Tephritidae , Animais , Feminino , Temperatura Alta , Oviposição , Temperatura , Tephritidae/genéticaRESUMO
Yolk processing pathways vary in the oocytes of benthophil and pelagophil teleosts. The present study investigated the yolk processing pattern in the oocytes of the fresh water catfish Clarias gariepinus at vitellogenic, maturation, and ovulated stages. This study concludes that during maturation stage, an electrophoretic shift in the major peptide band on Polyacrylamide gel electrophoresis (PAGE) occurs due to a decrease in the size of the yolk protein. The PMF spectrum of corresponding peptides from vitellogenic and ovulated oocytes revealed a difference in the minor ions. A minor difference in the molecular weight of the corresponding peptides occurs due to a difference in their amino acid composition. Maximal activity of the proteases cathepsin D and cathepsin B was observed in the vitellogenic oocytes, thus confirming their role in the processing of yolk. A significant transient increase in the activity of cathepsin B in the mature oocytes also suggests its role in oocyte maturation.
Assuntos
Peixes-Gato , Catepsina B , Catepsina D/metabolismo , Animais , Peixes-Gato/metabolismo , Catepsina B/metabolismo , Proteínas do Ovo/metabolismo , Oócitos/metabolismo , ProteóliseRESUMO
1. Phosvitin, a major phosphoprotein found in egg yolk, has strong antioxidant activity. Activation of elastase, collagenase, and hyaluronidase by reactive oxygen species are related to the degradation of ECM and skin aging. The objective of this study was to determine the anti-elastase and anti-hyaluronidase activity of phosvitin.2. Elastase from porcine pancreas and hyaluronidase from bovine testes were used to study the inhibitory activity of phosvitin. To elucidate the mechanism of enzyme inhibition, a Lineweaver-Burk plot was constructed.3. Phosvitin inhibited elastase and hyaluronidase activity in a dose-dependent manner. The IC50 value of phosvitin was 31.6 µg/ml and 1,270 µg/ml against elastase and hyaluronidase, respectively. The analysis of elastase and hyaluronidase kinetics indicated that the apparent Michaelis constant (appKm) was increased by phosvitin but the Vmax value was not affected.4. In conclusion, phosvitin exhibited competitive inhibitory activity against elastase and hyaluronidase. Thus, phosvitin could be used as a natural anti-aging agent in the cosmetics industry.
Assuntos
Gema de Ovo , Fosvitina , Animais , Bovinos , Galinhas , Feminino , Hialuronoglucosaminidase , SuínosRESUMO
This study examined the expression and role of vitellogenin (Vg) in the body of the firebug Pyrrhocoris apterus (Heteroptera, Insecta) during infection elicited by two entomopathogenic organisms, the nematode Steinernema carpocapsae and the fungus Isaria fumosorosea Infection by S. carpocapsae significantly upregulated Vg mRNA expression in the male body. The corresponding increase in Vg protein expression was also confirmed by electrophoretic and immunoblotting analyses. Remarkably, in females, the opposite tendency was noted. Nematodal infection significantly reduced both Vg mRNA and Vg protein expression levels in fat body and hemolymph, respectively. We speculate that infection of reproductive females reduces Vg expression to a level that is still sufficient for defense, but is insufficient for reproduction. This circumstance reduces energy expenditure and helps the individual to cope with the infection. Importantly, purified Vg significantly inhibited growth of Xenorhabdus spp., an entomotoxic bacteria isolated from S. carpocapsae. However, the effect of Vg against I. fumosorosea was not so obvious. The fungus significantly stimulated Vg gene expression in males; however, a similar increase was not recapitulated at the protein level. Nevertheless, in females, both mRNA and protein Vg levels were significantly reduced after the fungal infection. The obtained data demonstrate that Vg is probably an important defense protein, possibly with a specific activity. This considerably expands the known spectrum of Vg functions, as its primary role was thought to be limited to regulating egg development in the female body.
Assuntos
Heterópteros/genética , Interações Hospedeiro-Patógeno/fisiologia , Hypocreales/fisiologia , Proteínas de Insetos/genética , Rabditídios/fisiologia , Vitelogeninas/genética , Animais , Feminino , Expressão Gênica , Heterópteros/metabolismo , Heterópteros/microbiologia , Heterópteros/parasitologia , Interações Hospedeiro-Parasita , Interações Hospedeiro-Patógeno/genética , Proteínas de Insetos/metabolismo , Masculino , Vitelogeninas/metabolismoRESUMO
BACKGROUND: In order to supply adequate dietary protein for chronic kidney disease (CKD) patients while simultaneously controlling phosphorus intake, a novel method was developed for the preparation of low-phosphorus egg yolk protein (LPYP) using alkaline protease auxiliary dephosphorization. In addition, the physicochemical properties of LPYP were studied. RESULTS: In comparison with raw egg yolk protein (RYP) and defatted egg yolk protein (DFYP), LPYP was found to exhibit differences in amino acid (AA) composition, protein secondary structure, surface hydrophobicity, solubility and emulsion stability. It was observed that dephosphorization improved the AA composition, soluble protein content and dissolution stability of egg yolk protein. In addition, phosphate groups were found to impose a critical influence on the emulsion stability and particle size distribution. The final phosphorus to protein mass ratio (P/Pro) of LPYP was 5.64, which met the requirements of a protein diet for CKD patients. The FAO/WHO mode closeness and stability coefficient were 0.958 and 98.62% respectively. CONCLUSION: LPYP can be effectively obtained by alkaline protease hydrolysis and subsequent alkali dephosphorization. The prepared LPYP can be considered to be a type of safe and suitable protein resource for CKD patients. © 2018 Society of Chemical Industry.
Assuntos
Proteínas do Ovo/química , Gema de Ovo/química , Manipulação de Alimentos/métodos , Fósforo/análise , Aminoácidos/análise , Animais , Proteínas de Bactérias/química , Biocatálise , Galinhas , Ovos/análise , Endopeptidases/química , SolubilidadeRESUMO
The nutritional environment is crucial for Drosophila oogenesis in terms of controlling hormonal conditions that regulate yolk production and the progress of vitellogenesis. Here, we discovered that Drosophila Endophilin B (D-EndoB), a member of the endophilin family, is required for yolk endocytosis as it regulates membrane dynamics in developing egg chambers. Loss of D-EndoB leads to yolk content reduction, similar to that seen in yolkless mutants, and also causes poor fecundity. In addition, mutant egg chambers exhibit an arrest at the previtellogenic stage. D-EndoB displayed a crescent localization at the oocyte posterior pole in an Oskar-dependent manner; however, it did not contribute to pole plasm assembly. D-EndoB was found to partially colocalize with Long Oskar and Yolkless at the endocytic membranes in ultrastructure analysis. Using an FM4-64 dye incorporation assay, D-EndoB was also found to promote endocytosis in the oocyte. When expressing the full-length D-endoB(FL) or D-endoB(ΔSH3) mutant transgenes in oocytes, the blockage of vitellogenesis and the defect in fecundity in D-endoB mutants was restored. By contrast, a truncated N-BAR domain of the D-EndoB only partially rescued these defects. Taken together, these results allow us to conclude that D-EndoB contributes to the endocytic activity downstream of Oskar by facilitating membrane dynamics through its N-BAR domain in the yolk uptake process, thereby leading to normal progression of vitellogenesis.
Assuntos
Aciltransferases/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Gema de Ovo/citologia , Endocitose , Oócitos/citologia , Aciltransferases/química , Aciltransferases/genética , Animais , Membrana Celular/metabolismo , Polaridade Celular/genética , RNA Helicases DEAD-box/metabolismo , Proteínas de Drosophila/química , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/ultraestrutura , Gema de Ovo/metabolismo , Feminino , Fertilidade/genética , Perfilação da Expressão Gênica , Mutação/genética , Oócitos/metabolismo , Oócitos/ultraestrutura , Isoformas de Proteínas/metabolismo , Estrutura Terciária de Proteína , Transporte Proteico , Transdução de Sinais/genéticaRESUMO
Vitellogenenesis is a physiological process common in oviparous animals. The molecular profile, modifications, and utilization of vitellogenin (VTG), a precursor of yolk protein, have been characterized in various taxa to understand oogenesis within different modes of reproduction. Hormonal regulation of VTGs has been investigated in invertebrates, such as insects and crustaceans; conversely, little is known for cephalopods. In this study, we isolated two VTG genes (ue-VTG1 and ue-VTG2) from the loliginid swordtip squid, Uroteuthis edulis, via a comprehensive survey of a transcriptome database and subsequent cDNA cloning. Structural analysis of the two ue-VTGs revealed their unique features, namely the absence of two domains usually found in VTGs from other organisms: the von Willebrand factor D domain (vWD) and the domain of unknown function 1943 (DUF1943). Levels of ue-VTG1 and ue-VTG2 transcripts in the ovary, specifically in follicular cells, increased during the late-vitellogenic phase, suggesting that yolk accumulation progresses via paracrine interactions involving follicular cells and oocytes. N-terminal amino acid sequencing of biochemically purified yolk protein revealed its origins from these two VTGs, indicating that both are functional precursors of yolk protein. These results provide information that is essential to understanding the physiological pathway of yolk synthesis, accumulation, and storage in loliginid squids.
Assuntos
Decapodiformes , Regulação da Expressão Gênica/fisiologia , Oócitos/metabolismo , Vitelogênese/fisiologia , Vitelogeninas , Animais , Clonagem Molecular , Decapodiformes/genética , Decapodiformes/metabolismo , Feminino , Vitelogeninas/biossíntese , Vitelogeninas/genéticaRESUMO
Zebrafish phosvitin-derived peptide Pt5, consisting of the C-terminal 55 residues of phosvitin, has been shown to have an antimicrobial-immunomodulatory activity comparable to phosvitin. Here, we showed clearly that Pt5 had the capacity to inhibit tyrosinase (TYR) activity and melanin biosynthesis, and this inhibition was independent of cell proliferation and cytotoxic effects. Incubation of fluorescein isothiocyanate (FITC)-labeled Pt5 with B16F10 melanoma cells revealed that Pt5 was localized in the cytoplasm of the cells. In addition, Pt5 inhibited the expression of TYR, tyrosinase-related protein-1 (TRP-1), tyrosinase-related protein-2 (TRP-2), and microphthalmia-associated transcription factor (MITF) in B16F10 melanoma cells and reduced the intracellular cyclic adenosine monophosphate (cAMP) concentration in the cells, but it did not affect the cellular contents of pERK1/2 and ß-catenin, suggesting that Pt5 regulates melanin biosynthesis via cAMP signaling pathway rather than Wnt and MAPK pathways. Collectively, these data indicate that Pt5 has the potential to be used as a melanogenesis inhibitor in medical and cosmetic industry, a novel role ever reported.
Assuntos
AMP Cíclico/metabolismo , Melaninas/biossíntese , Fragmentos de Peptídeos/farmacologia , Fosvitina/farmacologia , Proteínas de Peixe-Zebra/farmacologia , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Oxirredutases Intramoleculares/metabolismo , Camundongos , Monofenol Mono-Oxigenase/metabolismo , Oxirredutases/metabolismo , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacosRESUMO
The provision of yolk precursor proteins to the oviparous egg is crucial for normal embryo development. In Echinodermata, a transferrin-like yolk component termed major yolk protein (MYP) is a major precursor protein in Echinoidea and Holothuroidea. In contrast, in Asteroidea a single vitellogenin (Vtg) was recently identified, but its role as primary yolk protein remains unclear. To resolve the apparent MYP-Vtg dichotomy in sea stars and to understand the dynamics of candidate yolk protein gene expression during the reproductive cycle, we investigated the molecular structures of sea star Vtg and MYP and quantified their transcript levels during oogenesis. By combining protein sequencing of the predominant proteins in ovulated eggs of Patiriella regularis with ovarian transcriptome sequencing and molecular cloning, we characterized two cDNAs encoding two bona fide Vtgs (PrVtg1 and PrVtg2) and a partial cDNA encoding MYP (PrMYP). PrMYP mRNA was found in low abundance in growing oocytes, possibly as maternal transcripts for translation after ovulation. In contrast, PrVtg transcripts, whose levels varied during the reproductive cycle, were not found in developing oocytes - rather, they were detected in ovarian follicle cells and pyloric caeca, indicating an extra-oocytic origin. Vtg accumulating in oocytes was stored in the form of cleaved products, which constituted the most abundant yolk polypeptides in ovulated sea star eggs; their levels decreased during early embryonic and larval development. Together, these traits are the hallmarks of a classical yolk protein - and hence, we contend that Vtg, and not MYP, is the main yolk protein in asteroids.
Assuntos
Proteínas do Ovo/genética , Estrelas-do-Mar/genética , Vitelogeninas/genética , Sequência de Aminoácidos , Animais , Clonagem Molecular , Sequência Conservada , Proteínas do Ovo/metabolismo , Embrião não Mamífero , Feminino , Expressão Gênica , Oócitos/fisiologia , Ovário/fisiologia , Filogenia , Pepinos-do-Mar/genética , Ouriços-do-Mar/genética , Estrelas-do-Mar/embriologia , Vitelogeninas/metabolismoRESUMO
To gain a better understanding of how corals form their eggs at both the molecular and cellular levels, we performed a differential screen (suppression subtractive hybridization) to identify genes related to oocyte development in a stony coral, Euphyllia ancora. Through the course of screening, a novel gene that contains three alternate repeats of fibronectin domain 2 and epidermal growth factor (EGF)-like domains, as well as an additional calcium-binding EGF-like domain (EGF-CA), was identified and tentatively named euphy after the scientific name of the coral, E. ancora. Quantitative RT-PCR revealed that expression levels of euphy increased in female colonies as the coral approached reproductive season. Tissue distribution analysis followed by mRNA in situ hybridization revealed that euphy is highly expressed in the ovarian (mesenterial) somatic cells in the body of E. ancora. Staining of tissue sections with an antibody against euphy protein (Euphy) revealed Euphy immunoreactivity in both ovarian somatic cells and oocytes. Subsequent Western blotting demonstrated the presence of abundant Euphy in unfertilized mature eggs. These results indicate that Euphy produced in the ovarian somatic cells is transported to and accumulates within oocytes as a yolk protein during oogenesis. We previously showed that two major yolk proteins, vitellogenin and egg protein, are similarly produced by ovarian somatic cells. Hence, the present study uncovered the third ovarian somatic-derived yolk protein in corals. Our data provide new information that contributes to a more comprehensive understanding of coral egg formation.
Assuntos
Antozoários/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Oócitos/metabolismo , Ovário/citologia , Ovário/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Ligação ao Cálcio/genética , DNA Complementar/biossíntese , DNA Complementar/genética , Fator de Crescimento Epidérmico/metabolismo , Feminino , Fibronectinas/metabolismo , Imuno-Histoquímica , Dados de Sequência Molecular , RNA Mensageiro/biossíntese , Reprodução , Distribuição TecidualRESUMO
Antioxidants, or anti-oxidant agents, have attracted a great deal of attention in recent years because of their roles in prevention of chronic diseases and utilization as preservatives in food and cosmetics. In this study, we clearly demonstrated that zebrafish recombinant phosvitin (rPv) is an antioxidant agent capable of inhibiting the oxidation of the linoleic acid, and scavenging the 2,2-diphenyl-1-picrylhydrazyl radical. We also showed that zebrafish rPv is a cellular antioxidant capable of protecting radical-mediated oxidation of cellular biomolecules. Importantly, zebrafish rPv is non-cytotoxic to murine macrophage RAW264.7 cells. It is the first report that showed the antioxidant activities of Pv in fishes, suggesting that zebrafish Pv can be an important antioxidant, which can be used as preservatives in food and cosmetics and even as supplementary mediator in different diseased states.
Assuntos
Antioxidantes/metabolismo , Fosvitina/metabolismo , Peixe-Zebra/metabolismo , Animais , Linhagem Celular , CamundongosRESUMO
The vitellogenin receptor (VgR) functions as an essential component in uptaking and transporting vitellogenin (Vg) in female adults, which is involved in ovary development and oviposition. This study aimed to clarify the molecular characteristics and function of VgR in the oriental fruit fly Bactrocera dorsalis (Hendel). Here, we identified the full-length of BdVgR (GenBank Accession No. JX469118), encoding a 1925 residue (aa) protein with a 214.72 kDa molecular mass and several typical motifs of low-density lipoprotein receptor superfamily (LDLR). Phylogenic analysis suggested that BdVgR was evolutionary conserved with other Dipteran VgRs. The expression of BdVgR was exclusively detected in the ovaries rather than head, thorax or other tissues. The developmental expression patterns showed that the signal of BdVgR was detectable in very beginning of adult stage, and positively correlated with the growth rate of ovaries and the expression levels of its ligands. In addition, we also demonstrated that the expression level of BdVgR, and ovary development were significantly suppressed after being injected with BdVgR-targeted dsRNA. Together, all of these results indicated that BdVgR was critical for yolk protein absorption and ovary maturation in B. dorsalis, playing a vital role in female reproduction.
Assuntos
Proteínas do Ovo/metabolismo , Proteínas de Insetos/metabolismo , Receptores de Superfície Celular/metabolismo , Tephritidae/crescimento & desenvolvimento , Vitelogeninas/metabolismo , Animais , Proteínas do Ovo/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Filogenia , Interferência de RNA , RNA Interferente Pequeno/genética , Receptores de Superfície Celular/genética , Tephritidae/genética , Tephritidae/metabolismoRESUMO
A one-step organic solvent lipid extraction method was used to separate lipids from spray-dried egg yolk. Organic solvents tested were chloroform:methanol (CM, 2:1, v:v), methyl-tert-butyl ether (MTBE), or hexane:isopropanol (HI, 3:2, v:v). The resulting defatted egg yolk powder had between 21 and 30% more protein and between 22 and 25% less lipid than the initial spray-dried egg yolk powder (p < 0.05). The solubility of the powder decreased from 20% to 4% (p < 0.05) when CM was used as the organic solvent, likely due to protein denaturation by the chloroform. Gels made from MBTE and HI-extracted protein concentrates had similar hardness (p > 0.05) and were both harder than gels made using the initial egg yolk powder (p < 0.05). MTBE gels were springier, more cohesive, and gummier (p < 0.05) with similar resistance to the initial egg yolk powder (p > 0.05). The results of this study showed that the functionality of the protein in the defatted egg yolk powder was best retained when MTBE was used as the lipid extraction solvent.
RESUMO
Caffeine (1,3,7-trimethylxanthine) is a widely consumed bioactive substance worldwide. Our recent study showed that a reduction in both reproduction and yolk protein production (vitellogenesis) caused by caffeine intake were improved by vitamin B12 supplementation, which is an essential co-factor in methionine metabolism. In the current study, we investigated the role of methionine in the reproduction of caffeine-ingested animals (CIAs). We assessed the effect of methionine metabolism on CIAs and found that caffeine intake decreased both methionine levels and essential enzymes related to the methionine cycle. Furthermore, we found that the caffeine-induced impairment of methionine metabolism decreased vitellogenesis and increased germ cell apoptosis in an LIN-35/RB-dependent manner. Interestingly, the increased germ cell apoptosis was restored to normal levels by methionine supplementation in CIAs. These results indicate that methionine supplementation plays a beneficial role in germ cell health and offspring development by regulating vitellogenesis.
Assuntos
Caenorhabditis elegans , Metionina , Animais , Metionina/farmacologia , Metionina/metabolismo , Cafeína/farmacologia , Cafeína/metabolismo , Apoptose , Células Germinativas , Racemetionina/metabolismo , Suplementos NutricionaisRESUMO
To improve the thermal gel properties of egg yolk, the effect of several valence metal ions (K+, Ca2+, Mg2+ and Fe3+) with different concentrations (0-0.72%) on the rheological, gel, and structural properties of egg yolk were investigated. Results showed that monovalent and divalent ions were beneficial to the formation of uniform and dense gel network, especially with the addition of 0.72% magnesium ion, which further improved gel hardness, water holding capacity (WHC) and viscoelastic properties, the properties of egg yolk gel increased with the increase of the concentration of mono-bivalent metal ions. Adding ferric ion remarkably increased the average particle size (d4,3) and apparent viscosity of egg yolk, destroying the disulfide bonds and the hydrophobic interactions in gel. Fourier transform infrared spectroscopy (FT-IR) and fluorescence spectra analysis revealed that metal ions promoted the hydrophobic aggregation among egg yolk proteins and induced the transition of protein secondary structure from ordered to disordered. This work will provide a theoretical reference for the development of low salt and nutrient fortified egg yolk products.
Assuntos
Galinhas , Gema de Ovo , Gema de Ovo/química , Animais , Géis/química , Cátions Bivalentes , Reologia , ViscosidadeRESUMO
To investigate the alterations of yolk protein during embryonic development in Wanxi white goose, the egg yolk protein composition at days 0, 4, 7, 14, 18, and 25 of incubation (D0, D4, D7, D14, D18, and D25) was analyzed by two-dimensional gel electrophoresis combined with mass spectrometry. A total of 65 spots representing 11 proteins with significant abundance changes were detected. Apolipoprotein B-100, vitellogenin-1, vitellogenin-2-like, riboflavin-binding protein, and serotransferrin mainly participated in nutrient (lipid, riboflavin, and iron ion) transport, and vitellogenin-2-like showed a lower abundance after D14. Ovomucoid-like were involved in endopeptidase inhibitory activity and immunoglobulin binding and exhibited a higher expression after D18, suggesting a potential role in promoting the absorption of immunoglobulin and providing passive immune protection for goose embryos after D18. Furthermore, myosin-9 and actin (ACTB) were involved in the tight junction pathway, potentially contributing to barrier integrity. Serum albumin mainly participated in cytolysis and toxic substance binding. Therefore, the high expression of serum albumin, myosin-9, and ACTB throughout the incubation might protect the developing embryo. Apolipoprotein B-100, vitellogenin-1, vitellogenin-2-like, riboflavin-binding protein, and serotransferrin might play a crucial role in providing nutrition for embryonic development, and VTG-2-like was preferentially degraded/absorbed.
Assuntos
Gansos , Vitelogeninas , Animais , Vitelogeninas/análise , Gansos/metabolismo , Apolipoproteína B-100/análise , Apolipoproteína B-100/metabolismo , Proteômica , Transferrina , Proteínas do Ovo/química , Desenvolvimento Embrionário , Albumina Sérica/metabolismo , Imunoglobulinas/análise , Miosinas/análise , Miosinas/metabolismo , Gema de Ovo/químicaRESUMO
Reproduction is an energy-intensive process requiring systemic coordination. However, the inter-organ signaling mechanisms that relay nutrient status to modulate reproductive output are poorly understood. Here, we use Drosophila melanogaster as a model to establish the integrated stress response (ISR) transcription factor, Atf4, as a fat tissue metabolic sensor that instructs oogenesis. We demonstrate that Atf4 regulates lipase activity to mediate yolk lipoprotein synthesis in the fat body. Depletion of Atf4 in the fat body also blunts oogenesis recovery after amino acid deprivation and re-feeding, suggestive of a nutrient-sensing role for Atf4. We also discovered that Atf4 promotes secretion of a fat-body-derived neuropeptide, CNMamide, which modulates neural circuits that promote egg-laying behavior (ovulation). Thus, we posit that ISR signaling in fat tissue acts as a "metabolic sensor" that instructs female reproduction-directly by impacting yolk lipoprotein production and follicle maturation and systemically by regulating ovulation.
Assuntos
Drosophila melanogaster , Oogênese , Animais , Feminino , Drosophila melanogaster/fisiologia , Oogênese/fisiologia , Ovulação , Oócitos , LipoproteínasRESUMO
This study investigated the impact of varied pH treatments on the structural, emulsification, and interfacial adsorption properties of egg yolk. The solubility of egg yolk proteins decreased and then increased in response to pH changes, with a minimum value (41.95 %) observed at pH 5.0. The alkaline condition (pH 9.0) significantly impacted the secondary/tertiary structure of egg yolk, with the yolk solution displaying the lowest surface tension value (15.98 mN/m). Emulsion stability was found to be optimal when egg yolk was used as the stabilizer at pH 9.0, which corresponded to the more flexible diastolic structure, smaller emulsion droplets, increased viscoelasticity, and enhanced resistance to creaming. At pH 9.0, proteins exhibited a maximum solubility (90.79 %) due to their unfolded conformation, yet the protein adsorption content at the oil-water interface showed relatively low (54.21 %). At this time, electrostatic repulsion between the droplets and the spatial site barrier made by proteins that were unable to efficiently adsorb at the oil-water interface kept the emulsion stable. Moreover, it was found that different pH treatments could effectively regulate the relative adsorption contents of various protein subunits at the oil-water interface, and all proteins except livetin displayed good interfacial adsorption capacity at the oil-water interface.