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1.
Artigo em Inglês | MEDLINE | ID: mdl-32058017

RESUMO

The vitellogenin receptor (VgR) plays a critical role in egg development by mediating endocytosis of the major yolk protein precursor vitellogenin (Vg). Therefore, identifying the VgR of beneficial insects and its characterization could lead to the development of novel egg production strategies to enhance their commercial values. Here, we present the cloning, expression, and functional characterization of the VgR from an economically important eri silkworm, Samia ricini. The complete mRNA sequence was 6002 bp with an ORF of 5484 bp, encoding a protein of 1827 amino acids. Sequence analyses revealed that the SrVgR contained all of the conservative structural motifs characteristics of LDLR family members. The SrVgR was specifically expressed in the ovary, and the mRNA level increased steadily in pupal stages, reached its peak on day 9, and then declined to a bare minimum in adults. RNA interference (RNAi) clearly reduced the VgR transcript levels, disrupted the ovarian development resulting in malformed ovarioles and abnormal development of eggs. Taken together, these data provide conclusive evidence for the essential roles of VgR in insect reproduction.


Assuntos
Bombyx/metabolismo , Proteínas do Ovo/metabolismo , Proteínas de Insetos/metabolismo , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Bombyx/genética , Clonagem Molecular , Proteínas do Ovo/genética , Feminino , Proteínas de Insetos/genética , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/genética , Homologia de Sequência de Aminoácidos
2.
J Agric Food Chem ; 68(4): 1157-1167, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31917922

RESUMO

To investigate the alterations of egg yolk protein abundances and their phosphorylation status at different storage temperatures, a comparative quantitative study of unfertilized chicken egg yolk after 15 days of storage at 4 and 37 °C was performed. Altogether, 445 proteins were identified in our study, of which the abundances of 154 proteins were significantly changed when comparing high-temperature storage with low-temperature storage, including 42 up-regulated and 112 down-regulated proteins. In the phosphoproteome, we identified a total of 137 phosphorylated sites on 326 peptides corresponding to 51 proteins. The results showed that the degree of phosphorylation for most egg yolk proteins was enhanced during high-temperature storage. Furthermore, GO analysis indicated that these phosphoproteins of egg yolk may be closely related to the binding, catalysis, and transport functions. The results provide further insights into the effect of storage temperature on egg proteome changes and their phosphorylation level. Moreover, this study can provide a theoretical basis for the improvement of egg quality during storage by phosphorylation modification in the food industry.


Assuntos
Proteínas do Ovo/química , Fosfoproteínas/química , Animais , Galinhas , Cromatografia Líquida de Alta Pressão , Proteínas do Ovo/metabolismo , Armazenamento de Alimentos , Espectrometria de Massas , Peptídeos/química , Peptídeos/metabolismo , Fosfoproteínas/metabolismo , Proteoma/química , Proteoma/metabolismo , Proteômica , Temperatura Ambiente
3.
Food Chem ; 311: 125998, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31874426

RESUMO

In this study we investigated the changes in the physicochemical properties, microstructure, protein structure, and intermolecular forces in egg yolk, plasma, and granule gels induced with a strong alkali. The results showed that egg yolks formed a three-dimensional gel, maintained by ionic and disulfide bonds from plasma and granules, respectively. According to spin-spin relaxation time, these gel systems fixed numerous water and lipid protons. Microstructure showed that egg yolks, plasma, and granules liberated their constituents on disruption, which randomly aggregated or bonded. Under a continuous strong alkali treatment, the second-derivative spectra of egg yolks and plasma appeared as an "aggregation band", while the ordered structure of granules decreased. The results suggested that the aggregation of alkali-induced egg yolks formed mainly due to structural changes in plasma proteins. Granules contributed to the increased hardness and the bonding of egg yolk gels via disulfide bonds induced by alkali treatment.


Assuntos
Álcalis/química , Proteínas do Ovo/química , Gema de Ovo/química , Animais , Varredura Diferencial de Calorimetria , Proteínas do Ovo/metabolismo , Géis/química , Microscopia Eletrônica de Transmissão , Estrutura Secundária de Proteína , Espectroscopia de Infravermelho com Transformada de Fourier
4.
Arch Insect Biochem Physiol ; 103(1): e21636, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31612557

RESUMO

As a member of the low-density lipoprotein receptor (LDLR) superfamily, vitellogenin (Vg) receptor (VgR) is responsible for the uptake of Vg into developing oocytes and is a potential target for pest control. Here, a full-length VgR complementary DNA (named as CsVgR) was isolated and characterized in the rice stem borer, Chilo suppressalis. The composite CsVgR gene contained an open reading frame of 5,484 bp encoding a protein of 1,827 amino acid residues. Structural analysis revealed that CsVgR contained two ligand-binding domains (LBDs) with four Class A (LDLRA ) repeats in LBD1 and seven in LBD2, which was structurally different from most non-Lepidopteran insect VgRs having five repeats in LBD1 and eight in LBD2. The developmental expression analysis showed that CsVgR messenger RNA expression was first detectable in 3-day-old pupae, sharply increased in newly emerged female adults, and reached a peak in 2-day-old female adults. Consistent with most other insects VgRs, CsVgR was exclusively expressed in the ovary. Notably, injection of dsCsVgR into late pupae resulted in fewer follicles in the ovarioles as well as reduced fecundity, suggesting a critical role of CsVgR in female reproduction. These results may contribute to the development of RNA interference-mediated disruption of reproduction as a control strategy of C. suppressalis.


Assuntos
Proteínas do Ovo/genética , Mariposas/genética , Receptores de Superfície Celular/genética , Animais , Proteínas do Ovo/química , Proteínas do Ovo/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/metabolismo , Mariposas/crescimento & desenvolvimento , Mariposas/metabolismo , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Filogenia , Interferência de RNA , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/química , Receptores de Superfície Celular/metabolismo , Análise de Sequência de Proteína
5.
J Insect Sci ; 19(6)2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31812980

RESUMO

Vitellogenin receptor (VgR) mediates the intake of vitellin via oocytes, thus exerting an important role in vitellogenesis. In this study, reverse transcription-polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends techniques were adopted to clone the CiVgR gene, namely the VgR gene of Calliptamus italicus, i.e., Orthopteran. The full length of CiVgR was 5,589 bp, and the open reading frame was estimated to be 5,265 bp, which encoded 1,754 amino acids (aa). Sequence alignment analysis showed that CiVgR belonged to the superfamily of low-density lipoprotein receptor genes, which contained several conserved domains, including ligand-binding domains, epidermal growth factor precursor homology domains, transmembrane domains, and cytoplasmic domains. However, no O-linked sugar domain was identified. Phylogenetic analysis showed that CiVgR had the closest genetic relationship to Blattarias. RT-PCR showed that CiVgR was only specifically expressed in the ovarian tissue of females. quantitative real time polymerase chain reaction showed that the transcription of CiVgR already appeared in the fourth-instar nymph of C. italicus, which gradually increased after adult emergence, peaked at the previtellogenesis stage, and then started to decrease. The expression pattern of CiVgR was closely associated with vitellogenesis. The findings of this study further our understanding of the molecular mechanisms involved in the reproduction of C. italicus, and provide new ideas to control this insect.


Assuntos
Proteínas do Ovo/metabolismo , Gafanhotos/metabolismo , Receptores de Superfície Celular/metabolismo , Sequência de Aminoácidos , Animais , Proteínas do Ovo/genética , Feminino , Masculino , Filogenia , Receptores de Superfície Celular/genética , Reprodução , Homologia de Sequência de Aminoácidos
6.
J Agric Food Chem ; 67(48): 13353-13361, 2019 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-31682436

RESUMO

Protein phosphorylation plays an important role in protein structure and function. To investigate the role of egg protein phosphorylation in chicken embryonic development, a comparative and quantitative phosphoproteomic analysis of fertilized chicken egg white and yolk was performed during incubation. Overall, 215 phosphosites mapped onto 205 phosphopeptides corresponding to 100 phosphoproteins were identified. Among these phosphoproteins, 123 phosphosites from 62 egg proteins were found significantly changed (p < 0.05) at day 12 during incubation. Furthermore, GO analysis suggested that these differentially phosphorylated proteins were associated with various molecular functions, primarily including binding, molecular function regulator, and transport activity. Such findings in this study improved our understanding of the protein molecular functions involved in chicken embryonic development from a protein phosphorylation perspective.


Assuntos
Galinhas/metabolismo , Proteínas do Ovo/química , Óvulo/química , Fosfoproteínas/química , Animais , Galinhas/genética , Galinhas/crescimento & desenvolvimento , Proteínas do Ovo/metabolismo , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Fosfoproteínas/metabolismo , Fosforilação , Proteômica , Espectrometria de Massas em Tandem
7.
Chemosphere ; 226: 874-882, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31509916

RESUMO

The exposure and health effects of fluoride are an ongoing topic that has attracted worldwide attention. Fluoride exposure disturbs the testicular development, sexual hormone levels and spermatogenesis. However, as to whether fluoride interferes with acrosome formation which is essential for production of capable spermatozoa during spermatogenesis still remains unclear. The objective was to determine the effects of fluoride on the acrosome formation and to further elucidate the potential mechanism of impaired reproductive function. For this, forty adult rats were assigned into four groups. The control group received distilled water, while the other three groups were treated with 25, 50 and 100 mg NaF/L via drinking water for 56 d, respectively. Testes were processed for total RNA extraction and western blot analysis. Three samples of each group were fixed in 2.5% glutaraldehyde solution for transmission electron microscopy analysis. From the results, we first found that fluoride decreased the expression of mRNA and protein levels of Zpbp1, Spaca1 and Dpy19l2 of seven markers during acrosome biogenesis in testes. Furthermore, fluoride damaged not only the acrosome structure, but also the structure of the nuclear lamina which was observed to be discontinuous and partially missing by transmission electron microscopy. Moreover, the results indicated that the altered structure in nuclear lamina maybe due to reduced LMNB2 expression in testis induced by fluoride. In a nutshell, fluoride exposure could restrain acrosome biogenesis during spermatogenesis and contribute to the elucidation of the underlying mechanisms of fluoride-induced male reproductive toxicity.


Assuntos
Acrossomo/patologia , Fluoretos/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Espermatogênese/efeitos dos fármacos , Espermatozoides/patologia , Testículo/patologia , Acrossomo/efeitos dos fármacos , Acrossomo/metabolismo , Animais , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas de Plasma Seminal/genética , Proteínas de Plasma Seminal/metabolismo , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Testículo/efeitos dos fármacos , Testículo/metabolismo
8.
J Agric Food Chem ; 67(42): 11675-11683, 2019 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-31545598

RESUMO

To better appreciate the alterations of egg proteins and their modifications during embryonic development, a comparative and quantitative study was performed aimed at chicken egg white and yolk proteome and N-glycoproteome after 12 days of incubation using tandem mass tag (TMT)-labeling technology in conjunction with reversed-phase high-performance liquid chromatography (RP-HPLC). A total of 334 unique N-glycosite-containing peptides from 153 N-glycoproteins were identified, of which 82 N-glycosite-containing peptides showed significant changes after 12 days of incubation. The varied proteome was mainly involved with antibacterial, ionic binding, cell proliferation, and embryonic development, while the different degrading and/or absorbing priorities of egg proteins were proposed. This study provides substantial insight into the effects of N-glycoprotein variations on the utilization of egg proteins by chicken embryo during incubation.


Assuntos
Embrião de Galinha/química , Proteínas do Ovo/química , Glicoproteínas/química , Animais , Embrião de Galinha/crescimento & desenvolvimento , Embrião de Galinha/metabolismo , Galinhas/metabolismo , Cromatografia Líquida de Alta Pressão , Proteínas do Ovo/metabolismo , Clara de Ovo/química , Gema de Ovo/química , Gema de Ovo/metabolismo , Glicoproteínas/metabolismo , Proteômica , Espectrometria de Massas em Tandem
9.
Food Funct ; 10(9): 6074-6087, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31490476

RESUMO

Buffering capacity is a characteristic of foods to resist changes in pH, which is important to consider in gastric digestion as it will impact physicochemical breakdown of food. A standardized method to measure and quantify buffering capacity in the context of digestion is needed to improve in vitro digestion studies by providing a better estimation of acid secretions and subsequent protein digestibility. The objective of this study was to develop a method to measure buffering capacity in the context of digestion and develop a regression model to predict buffering capacity using protein-based model foods. Buffering capacity was analyzed by titrating 0.16 M HCl to egg and whey-protein based dispersions and gels of varying protein content and particle size and recording the pH after each addition. Calculated parameters from buffering capacity experiments included total acid added, area under the curve, total buffering capacity, relative [H+] increase, and lag phase. A regression model was developed to predict each buffering capacity parameter based on protein concentration, specific surface area, aspartic acid and glutamic acid content. Results showed that higher protein concentration and smaller surface area resulted in higher buffering capacity. A validation dataset was used to evaluate the goodness of fit of the model to the data with different protein concentrations, surface area or protein source. Results indicated that total buffering capacity and lag phase parameters can be used to quantify buffering capacity of protein gels in the context of digestion, since they provided a good fit to the observational and validation data sets.


Assuntos
Proteínas do Ovo/química , Estômago/química , Proteínas do Soro do Leite/química , Animais , Tampões (Química) , Bovinos , Galinhas , Digestão , Proteínas do Ovo/metabolismo , Mucosa Gástrica/metabolismo , Concentração de Íons de Hidrogênio , Modelos Biológicos , Proteínas do Soro do Leite/metabolismo
10.
PLoS Pathog ; 15(9): e1007924, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31487324

RESUMO

Parasitic helminths evade, skew and dampen human immune responses through numerous mechanisms. Such effects will likely have consequences for HIV-1 transmission and disease progression. Here we analyzed the effects that soluble egg antigen (SEA) from Schistosoma mansoni had on modulating HIV-1 infection and cytokine/chemokine production in vitro. We determined that SEA, specifically through kappa-5, can potently bind to DC-SIGN and thereby blocks DC-SIGN mediated HIV-1 trans-infection (p<0.05) whilst not interfering with cis-infection. DCs exposed to SEA whilst maturing under Th2 promoting conditions, will upon co-culture with naïve T-cells induce a T-cell population that was less susceptible to HIV-1 R5 infection (p<0.05) compared to DCs unexposed to SEA, whereas HIV-1 X4 virus infection was unaffected. This was not observed for DCs exposed to SEA while maturing under Th1 or Th1/Th2 (Tmix) promoting conditions. All T-cell populations induced by SEA exposed DCs demonstrate a reduced capacity to produce IFN-γ and MIP-1ß. The infection profile of T-cells infected with HIV-1 R5 was not associated with down-modulation of CCR5 cell surface expression. We further show that DCs maturing under Tmix conditions exposed to plant recombinant omega-1 protein (rω-1), which demonstrates similar functions to natural ω-1, induced T-cell populations that were less sensitive for HIV-1 R5 infection (p<0.05), but not for X4 virus infection. This inhibition associated again with a reduction in IFN-γ and MIP-1ß expression, but additionally correlated with reduced CCR5 expression. We have shown that SEA parasite antigens and more specifically rω-1 can modulate HIV-1 infectivity with the potential to influence disease course in co-infected individuals.


Assuntos
Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Proteínas do Ovo/imunologia , Infecções por HIV/metabolismo , Animais , Anticorpos Anti-Helmínticos/metabolismo , Antígenos de Helmintos/metabolismo , Linfócitos T CD4-Positivos/imunologia , Citocinas/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Proteínas do Ovo/metabolismo , Infecções por HIV/imunologia , Infecções por HIV/prevenção & controle , HIV-1/imunologia , Humanos , Ativação Linfocitária , Receptores CCR5/metabolismo , Schistosoma mansoni/metabolismo , Linfócitos T Auxiliares-Indutores/imunologia , Replicação Viral/imunologia
11.
J Agric Food Chem ; 67(35): 9950-9957, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31403788

RESUMO

Protein glycosylation is a ubiquitous posttranslational modification that modulates protein properties, thereby influencing bioactivities within a system. Duck egg white (DEW) proteins exhibit diverse biological properties compared with their chicken egg white (CEW) counterparts, which might be related to glycosylation. N-Glycoproteome analysis of DEW was conducted, and a total of 231 N-glycosites from 68 N-glycoproteins were identified. Gene ontology analysis was used to elucidate the biofunctions of DEW N-glycoproteins and compare them with those of CEW, which showed that the differences mostly involved molecular functions and biological processes. The biological functions of DEW N-glycoproteins were illuminated through bioinformatics analysis and comparison with CEW orthologues, which showed different allergenicities and antibacterial abilities. These divergences might be initiated by specific alterations in glycosylation, which can enhance the proteolysis resistance and protein steric hindrance. These results provide new insights for discovering the effects of N-glycosylation on biofunctions during the divergence of homologous proteins.


Assuntos
Galinhas/genética , Patos/genética , Proteínas do Ovo/química , Glicoproteínas/química , Animais , Evolução Biológica , Galinhas/metabolismo , Patos/metabolismo , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Clara de Ovo/química , Glicoproteínas/genética , Glicoproteínas/metabolismo , Glicosilação , Proteômica
12.
Int J Biol Macromol ; 138: 116-124, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31295501

RESUMO

In this paper, the relationship between protein structure changes and in vitro digestion of preserved egg white (PEW) during pickling was studied. Results showed that as the pickling time increased, moisture content of PEW exhibited a decreasing trend, pH value and hardness of PEW exhibited an increasing trend. Environmental Scanning Electron Microscopy (ESEM) revealed that alkali-treated duck egg white could form a more compact gel network structure during pickling. Transmission Electron Microscopy (TEM) showed that PEW consisted of two sorts of structures, namely fibrillar (from 7th to 21st day) and particulate (from 28th to 42nd day). Fourier Transform Infrared (FTIR) spectroscopy showed that there was an increase in the content of ß-sheets and a decrease in the content of α-helices during pickling. Protein digestibility of PEW was highest at day 14 and the lowest at day 42. Moreover, the content of ß-sheets was negatively correlated with protein digestibility. Peptides identification using LC-MS/MS highlighted that the number of different peptides were positively correlated with the pickling. The number of unique peptides was negatively correlated with protein digestibility at the end of gastric and intestinal digestion. This study would provide guidance for studying nutritional value and controlling quality of preserved eggs.


Assuntos
Digestão , Proteínas do Ovo/química , Clara de Ovo/química , Conservação de Alimentos , Fenômenos Químicos , Proteínas do Ovo/metabolismo , Estrutura Secundária de Proteína , Fatores de Tempo
13.
BMC Dev Biol ; 19(1): 14, 2019 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-31277577

RESUMO

BACKGROUND: Insulin/insulin-like growth peptide signaling (IIS) down-regulates hemolymph sugar level and facilitates larval growth in the soybean pod borer, Maruca vitrata. The objective of this study is to determine whether IIS of M. vitrata can mediate ovarian development of adult females. RESULTS: A pair of ovaries consists of 8 ovarioles, each of which is separated into distal germarium and proximal vitellarium in M. vitrata. In the germarium, oocyte development occurred with active mitotic activity which was visible by incorporating bromodeoxyribose uridine. Previtellogenic development and subsequent vitellogenesis began soon after adult emergence. They continued with increase of female age. Oocyte development was facilitated by up-regulation of vitellogenin (Vg) and Vg receptor (VgR) gene expression. Larval diets significantly influenced on ovarian development of M. vitrata because oocyte development varied with pupal size derived from larvae treated with different nutritional diets. Its ovarian development was dependent on endocrine signal(s) from the head because decapitation soon after adult emergence prevented oogenesis and subsequent vitellogenesis along with marked reduction of Vg and VgR expression. Topical application of juvenile hormone (JH) significantly recovered its ovarian development whereas farnesoic acid (a precursor of JH biosynthesis) or 20-hydroxyecdysone treatment did not. JH stimulated vitellogenesis and choriogenesis, but not previtellogenic development. In contrast, insulin injection to decapitated females stimulated oocyte differentiation and vitellogenesis along with increase of Vg and VgR expression. To further analyze the effect of insulin on ovarian development, expression of four IIS components (InR, FOXO, Akt, and TOR) genes was manipulated by RNA interference. Hemocoelic injection of gene-specific double stranded RNAs significantly reduced their target gene mRNA levels and interfered with ovarian development. An addition of insulin to JH treatment against decapitated females enhanced the gonadotropic effect of JH by stimulating oogenesis. CONCLUSIONS: IIS plays crucial role in mediating previtellogenic development of M. vitrata in response to nutrient signal. It also enhances the gonadotropic effect of JH II on vitellogenesis.


Assuntos
Fator de Crescimento Insulin-Like I/metabolismo , Insulina/metabolismo , Hormônios Juvenis/metabolismo , Ovário/crescimento & desenvolvimento , Vitelogênese/fisiologia , Animais , Ecdisterona/farmacologia , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Ácidos Graxos Insaturados/farmacologia , Feminino , Proteína Forkhead Box O1/genética , Mariposas , Oogênese/fisiologia , Proteínas Proto-Oncogênicas c-akt/genética , Interferência de RNA , RNA Interferente Pequeno/genética , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores de Somatomedina/genética , Serina-Treonina Quinases TOR/genética , Vitelogeninas/genética , Vitelogeninas/metabolismo
14.
Nat Commun ; 10(1): 2271, 2019 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118423

RESUMO

Following fertilization, cortical granules exocytose ovastacin, a metalloendopeptidase that cleaves ZP2 in the zona pellucida surrounding mouse eggs to prevent additional sperm binding. Using high- and super-resolution imaging with ovastacinmCherry as a fluorescent marker, we characterize cortical granule dynamics at single granule resolution in transgenic mouse eggs. Newly-developed imaging protocols provide an unprecedented view of vesicular dynamics near the plasma membrane in mouse eggs. We discover that cortical granule anchoring in the cortex is dependent on maternal MATER and document that myosin IIA is required for biphasic trafficking to the plasma membrane. We observe local clearance of cortical actin during exocytosis and determine that pharmacologic or genetic disruption of trafficking to the plasma membrane impairs secretion of cortical granules and results in polyspermy. Thus, the regulation of cortical granule dynamics at the cortex-plasma membrane interface is critical for exocytosis and the post-fertilization block to sperm binding that ensures monospermic fertilization.


Assuntos
Grânulos Citoplasmáticos/metabolismo , Exocitose/fisiologia , Interações Espermatozoide-Óvulo/fisiologia , Zona Pelúcida/metabolismo , Animais , Antígenos/metabolismo , Membrana Celular/metabolismo , Proteínas do Ovo/metabolismo , Feminino , Microscopia Intravital , Proteínas Luminescentes/química , Proteínas Luminescentes/genética , Masculino , Metaloproteases/química , Metaloproteases/genética , Metaloproteases/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia de Fluorescência , Glicoproteínas da Zona Pelúcida/metabolismo
15.
Parasit Vectors ; 12(1): 205, 2019 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-31060579

RESUMO

BACKGROUND: Vitellogenin (Vg), a key molecule for oocyte development synthesized in the fat body during blood-feeding, is released into the hemolymph and then taken into the oocytes via Vg receptor (VgR) in ticks. Previously, we showed that VgR mRNA is expressed in the ovary at the adult stage of parthenogenetic Haemaphysalis longicornis ticks and its expression increases after blood-feeding. However, intracellular localization of VgR mRNA and protein at each developmental stage of oocytes during oogenesis remains largely unclear. METHODS: mRNA and protein expression profiles of H. longicornis VgR (HlVgR) in the oocytes from the unfed to oviposition periods were analyzed by real-time PCR, in situ hybridization, and immunostaining. To elucidate the timing of the onset of Vg uptake, RNA interference (RNAi)-mediated gene silencing of HlVgR was performed. RESULTS: In situ hybridization revealed that HlVgR mRNA was detected in the cytoplasm of stage I-III oocytes, and weaker positive signals for HlVgR mRNA were found in the cell periphery of stage IV and V oocytes. Likewise, HlVgR protein was detected by immunostaining in the cytoplasm of stage I-III oocytes and in the cell periphery of stage IV and V oocytes. Each developmental stage of the oocytes showed distinct patterns of mRNA and protein expression of HlVgR. Moreover, RNAi of HlVgR caused delayed or arrested development in the oocytes. The ovaries of control ticks showed all developmental stages of oocytes, whereas stage I-III oocytes were found in the ovaries of HlVgR-RNAi ticks at 5 days after engorgement. CONCLUSIONS: These results suggest that active uptake of Vg is required for development from stage III to stage IV during oogenesis. Our data clearly revealed an apparent shift in the intracellular localization of VgR for both mRNA and protein level in oocytes during oogenesis.


Assuntos
Proteínas do Ovo/metabolismo , Ixodidae/metabolismo , Oogênese/fisiologia , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/metabolismo , Vitelogeninas/metabolismo , Animais , Proteínas do Ovo/genética , Feminino , Ixodidae/genética , Ixodidae/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos BALB C , Oócitos/crescimento & desenvolvimento , Oócitos/metabolismo , Oogênese/genética , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Receptores de Superfície Celular/genética , Transcriptoma
16.
Cell Mol Life Sci ; 76(11): 2133-2169, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30937469

RESUMO

To correctly transfer information, neuronal networks need to continuously adjust their synaptic strength to extrinsic stimuli. This ability, termed synaptic plasticity, is at the heart of their function and is, thus, tightly regulated. In glutamatergic neurons, synaptic strength is controlled by the number and function of AMPA receptors at the postsynapse, which mediate most of the fast excitatory transmission in the central nervous system. Their trafficking to, at, and from the synapse, is, therefore, a key mechanism underlying synaptic plasticity. Intensive research over the last 20 years has revealed the increasing importance of interacting proteins, which accompany AMPA receptors throughout their lifetime and help to refine the temporal and spatial modulation of their trafficking and function. In this review, we discuss the current knowledge about the roles of key partners in regulating AMPA receptor trafficking and focus especially on the movement between the intracellular, extrasynaptic, and synaptic pools. We examine their involvement not only in basal synaptic function, but also in Hebbian and homeostatic plasticity. Included in our review are well-established AMPA receptor interactants such as GRIP1 and PICK1, the classical auxiliary subunits TARP and CNIH, and the newest additions to AMPA receptor native complexes.


Assuntos
Proteínas de Transporte/metabolismo , Rede Nervosa/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Plasticidade Neuronal/fisiologia , Proteínas Nucleares/metabolismo , Receptores de AMPA/metabolismo , Animais , Proteínas de Transporte/genética , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Regulação da Expressão Gênica , Ácido Glutâmico/metabolismo , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas do Tecido Nervoso/genética , Neurônios/citologia , Neurônios/metabolismo , Proteínas Nucleares/genética , Transporte Proteico , Receptores de AMPA/genética , Sinapses/metabolismo , Transmissão Sináptica
17.
Food Chem ; 289: 694-700, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-30955667

RESUMO

The major allergen of chum salmon (Oncorhynchus keta) roe is the ß'-component (Onc k 5, ß'-c), which is a yolk protein and a fragment of vitellogenin. When yolk content containing ß'-c was orally administered to mice, ß'-c passed through the gastrointestinal tract and was excreted in feces without marked degradation. The direct administration of ß'-c to ligated jejunal and ileal loops showed that ß'-c was absorbed through the small intestine and transferred into the blood. Immunohistochemical staining showed that orally administered ß'-c was distributed from the apical side to the basal side of intestinal epithelial cells, suggesting that endocytosis may be involved in the intestinal absorption of ß'-c. In conclusion, ß'-c is absorbed along a large portion of the small intestine and circulates in the blood stream without significant digestion. The resistance of ß'-c to gastrointestinal digestion seems to contribute to its strong allergenicity.


Assuntos
Alérgenos/metabolismo , Proteínas de Peixes/metabolismo , Galectina 3/metabolismo , Trato Gastrointestinal/metabolismo , Salmão , Animais , Culinária , Digestão , Proteínas do Ovo/metabolismo , Células Epiteliais/metabolismo , Hipersensibilidade Alimentar , Intestino Delgado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR
18.
Parasit Vectors ; 12(1): 173, 2019 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-30992086

RESUMO

BACKGROUND: Schistosome parasites lay up to a thousand eggs per day inside the veins of their mammalian hosts. The immature eggs deposited by females against endothelia of venules will embryonate within days. Approximately 30% of the eggs will migrate to the lumen of the intestine to continue the parasite life-cycle. Many eggs, however, are trapped in the liver and intestine causing the main pathology associated with schistosomiasis mansoni and japonica, the liver granulomatous response. Excretory-secretory egg proteins drive much of egg-induced pathogenesis of schistosomiasis mansoni, and Schistosoma japonicum induce a markedly distinct granulomatous response to that of S. mansoni. METHODS: To explore the basis of variations in this responsiveness, we investigated the proteome of eggs of S. japonicum. Using mass spectrometry qualitative and quantitative (SWATH) analyses, we describe the protein composition of S. japonicum eggs secretory proteins (ESP), and the differential expression of proteins by fully mature and immature eggs, isolated from faeces and ex vivo adults. RESULTS: Of 957 egg-related proteins identified, 95 were exclusively found in S. japonicum ESP which imply that they are accessible to host immune system effector elements. An in-silico analysis implies that ESP are able of stimulating the innate and adaptive immune system through several different pathways. While quantitative SWATH analysis revealed 124 proteins that are differentially expressed by mature and immature S. japonicum eggs, illuminating some important aspects of eggs biology and infection, we also show that mature eggs are more likely than immature eggs to stimulate host immune responses. CONCLUSIONS: Here we present a list of potential targets that can be used to develop better strategies to avoid severe morbidity during S. japonicum infection, as well as improving diagnosis, treatment and control of schistosomiasis japonica.


Assuntos
Proteínas do Ovo/metabolismo , Proteínas de Helminto/metabolismo , Óvulo/metabolismo , Proteoma , Schistosoma japonicum/metabolismo , Animais , Sobrevivência Celular , Proteínas do Ovo/genética , Feminino , Perfilação da Expressão Gênica , Proteínas de Helminto/genética , Camundongos , Schistosoma japonicum/citologia
19.
Philos Trans R Soc Lond B Biol Sci ; 374(1767): 20180312, 2019 03 04.
Artigo em Inglês | MEDLINE | ID: mdl-30967014

RESUMO

It is known that plant arboviruses infect insect vector cells by endocytosis; however, the cellular receptors that mediate endocytosis have not been well defined. In our recently published work and this study, by clarifying the vertical transmission mechanism of Rice stripe virus (RSV) in Laodelphax striatellus, we provide a novel paradigm for how arboviruses enter insect germ-line cells. Instead of direct interaction with a viral receptor, the virus binds to a secreted ligand protein, hitchhiking the ligand-receptor pathway to achieve cell entry. Vitellogenin (Vg) is an indispensable protein for embryo development that is synthesized extra-ovarially and taken up by germ-line cells through Vg receptor (VgR)-mediated endocytosis. After revealing that RSV invades L. striatellus ovary by a specific molecular interaction with the insect Vg in haemolymph, this study addressed VgR's function in mediating the RSV invasion of the germarium nurse cells, further confirming the ligand's receptor-mediated viral cell-invasion mechanism. Understanding the viral ovary-entry pathways in vectors will help to find suitable measures to block the trans-generation transmission of the viruses. This article is part of the theme issue 'Biotic signalling sheds light on smart pest management'.


Assuntos
Hemípteros/microbiologia , Insetos Vetores/microbiologia , Oryza/virologia , Doenças das Plantas/microbiologia , Tenuivirus/fisiologia , Animais , Proteínas do Ovo/metabolismo , Feminino , Hemípteros/genética , Proteínas de Insetos/metabolismo , Insetos Vetores/genética , Ligantes , Ovário/microbiologia , Receptores de Superfície Celular/metabolismo
20.
Genetics ; 212(1): 213-229, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30867196

RESUMO

During meiosis, a series of evolutionarily conserved events allow for reductional chromosome division, which is required for sexual reproduction. Although individual meiotic processes have been extensively studied, we currently know far less about how meiosis is regulated and coordinated. In the Caenorhabditis elegans gonad, mitogen-activated protein kinase (MAPK) signaling drives oogenesis while undergoing spatial activation and deactivation waves. However, it is currently unclear how MAPK activation is governed and how it facilitates the progression of oogenesis. Here, we show that the oocyte and germline-related 2 (ogr-2) gene affects proper progression of oogenesis. Complete deletion of ogr-2 results in delayed meiotic entry and late spatial onset of double-strand break repair. Elevated levels of apoptosis are observed in this mutant, independent of the meiotic canonical checkpoints; however, they are dependent on the MAPK terminal member MPK-1/ERK. MPK-1 activation is elevated in diplotene in ogr-2 mutants and its aberrant spatial activation correlates with stages where meiotic progression defects are evident. Deletion of ogr-2 significantly reduces the expression of lip-1, a phosphatase reported to repress MPK-1, which is consistent with OGR-2 localization at chromatin in germ cells. We suggest that OGR-2 modulates the expression of lip-1 to promote the timely progression of meiosis through MPK-1 spatial deactivation.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Proteínas de Ciclo Celular/metabolismo , Proteínas do Ovo/metabolismo , Sistema de Sinalização das MAP Quinases , Meiose , Oogênese , Proteínas Tirosina Fosfatases/metabolismo , Animais , Caenorhabditis elegans/fisiologia , Proteínas de Caenorhabditis elegans/fisiologia , Proteínas do Ovo/fisiologia , Feminino
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