Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.336
Filtrar
1.
Zoolog Sci ; 38(5): 451-458, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34664920

RESUMO

Viviparous fish, including white-edged rockfish (Sebastes taczanowskii), accumulate substantial yolk mass in the oocytes; however, the details of the molecular mechanisms underlying yolk formation are not yet fully understood, especially concerning multiplicity in the yolk precursor vitellogenin (Vtg). The present study aimed to reveal the hepatic transcriptional profiles of multiple vtg gene transcripts (vtgAa, vtgAb, vtgC) during the reproductive cycle in captive female white-edged rockfish reared in an aquarium under natural photo-thermal conditions. The serum estradiol-17ß concentration and the hepatic transcript levels of all vtg subtypes increased with the progress of vitellogenesis; both levels decreased at the beginning of oocyte maturation and remained low during the gestation period. Considering the similarity in the transcriptional profiles of vtg subtypes between Sebastes and Oncorhynchus, along with the differences between Sebastes and Morone, it is suggested that the transcription patterns of multiple vtg genes relate to neither their reproductive modes (viviparity versus oviparity) nor to teleost phylogeny.


Assuntos
Fígado/metabolismo , Perciformes/fisiologia , Vitelogeninas/metabolismo , Animais , Estradiol/sangue , Feminino , Ovário/fisiologia , Transcriptoma , Vitelogênese , Vitelogeninas/genética
2.
Proc Natl Acad Sci U S A ; 118(39)2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34544864

RESUMO

It is well documented that the juvenile hormone (JH) can function as a gonadotropic hormone that stimulates vitellogenesis by activating the production and uptake of vitellogenin in insects. Here, we describe a phenotype associated with mutations in the Drosophila JH receptor genes, Met and Gce: the accumulation of mature eggs with reduced egg length in the ovary. JH signaling is mainly activated in ovarian muscle cells and induces laminin gene expression in these cells. Meanwhile, JH signaling induces collagen IV gene expression in the adult fat body, from which collagen IV is secreted and deposited onto the ovarian muscles. Laminin locally and collagen IV remotely contribute to the assembly of ovarian muscle extracellular matrix (ECM); moreover, the ECM components are indispensable for ovarian muscle contraction. Furthermore, ovarian muscle contraction externally generates a mechanical force to promote ovulation and maintain egg shape. This work reveals an important mechanism for JH-regulated insect reproduction.


Assuntos
Proteínas da Matriz Extracelular/metabolismo , Matriz Extracelular/metabolismo , Hormônios Juvenis/farmacologia , Oócitos/citologia , Oogênese , Ovulação , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Colágeno Tipo IV/genética , Colágeno Tipo IV/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster , Matriz Extracelular/efeitos dos fármacos , Proteínas da Matriz Extracelular/genética , Feminino , Laminina/genética , Laminina/metabolismo , Mutação , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Fatores de Transcrição/genética , Vitelogênese , Vitelogeninas/metabolismo
3.
Molecules ; 26(16)2021 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-34443500

RESUMO

Prostaglandins are a group of important cell-signaling molecules involved in the regulation of ovarian maturation, oocyte development, egg laying and associated behaviors in invertebrates. However, the presence of prostaglandin E2 (PGE2), the key enzymes for PGE2 biosynthesis and its interference by drugs were not investigated previously in the ovary of ticks. The present study was undertaken to assess the modulation of the PGE2-mediated pathway in the eclosion blocking effect of flumethrin and terpenoid subfraction isolated from Artemisia nilagirica in Rhipicephalus annulatus ticks. The acaricidal activities and chemical profiling of the terpenoid subfraction were performed. The localization of the cyclooxygenase1 (COX1) and prostaglandin E synthase (PGES) enzymes and the quantification of PGE2 in the ovaries of the ticks treated with methanol (control), flumethrin and terpenoid subfraction were also undertaken. In addition, the vitellogenin concentration in hemolymph was also assayed. Both flumethrin and the terpenoid subfraction of A. nilagirica elicited a concentration-dependent inhibition of fecundity and blocking of hatching of the eggs. The COX1 could not be detected in the ovaries of treated and control ticks, while there was no significant difference observed in the concentration of vitellogenin (Vg) in them. The presence of PGES in the oocytes of control ticks was confirmed while the immunoreactivities against PGES were absent in the vitellogenic oocytes of ticks treated with flumethrin and terpenoid subfraction. The levels of PGE2 were below the detection limit in the ovaries of the flumethrin-treated ticks, while it was significantly lower in the ovaries of the terpenoid subfraction-treated ticks. Hence, the prostaglandin E synthase and PGE2 were identified as very important mediators for the signaling pathway for ovarian maturation and oviposition in ticks. In addition, the key enzyme for prostaglandin biosynthesis, PGES and the receptors for PGE2 can be exploited as potential drug targets for tick control. The detection of PGES by immunohistochemistry and quantification of PGE2 by LC-MSMS can be employed as valuable tools for screening newer compounds for their eclosion blocking acaricidal effects.


Assuntos
Artemisia/química , Dinoprostona/metabolismo , Piretrinas/farmacologia , Rhipicephalus/efeitos dos fármacos , Terpenos/isolamento & purificação , Terpenos/farmacologia , Animais , Anticorpos/metabolismo , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Hemolinfa/metabolismo , Imersão , Ovário/efeitos dos fármacos , Ovário/enzimologia , Peroxidase/metabolismo , Prostaglandina-E Sintases/metabolismo , Vitelogeninas/metabolismo
4.
Environ Toxicol Pharmacol ; 87: 103720, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34332080

RESUMO

Silica nanoparticles (SiNPs) and cadmium chloride (CdCl2) are two important environmental pollutants. In previous research, found that SiNPs in zebrafish larvae can amplify the cardiovascular damage caused by cadmium. Whether SiNPs in the ovaries can amplify the adverse effects of cadmium on the zebrafish ovaries is worth studying problem. In this study, sexually mature female zebrafish were used as model organisms and exposed to 1 µmol/L CdCl2 and/or 25 µg/mL SiNPs for 30 days. The results showed that the structure and function of ovaries in the sole and combined exposure groups changed significantly, resulting in reduced ovarian quality, decreased number of mature oocytes, and the development of malformed offspring. A deep-sequencing analysis showed that organisms' lipid metabolism and transportation, estrogen metabolism, and response to the maturation, meiosis, and vitellogenin synthesis of oocytes were significantly affected by single exposure or combined exposure. These findings provide further insights into the harm of cooperation of CdCl2 and/or SiNPs to the aquatic ecosystems.


Assuntos
Cloreto de Cádmio/toxicidade , Nanopartículas/toxicidade , Ovário/efeitos dos fármacos , Dióxido de Silício/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Embrião não Mamífero , Estrogênios/metabolismo , Feminino , Metabolismo dos Lipídeos/efeitos dos fármacos , Masculino , Meiose/efeitos dos fármacos , Ovário/metabolismo , Ovário/patologia , Transcriptoma/efeitos dos fármacos , Vitelogeninas/metabolismo , Peixe-Zebra
5.
Arch Insect Biochem Physiol ; 108(2): e21836, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34288123

RESUMO

In Asian rice systems, Cyrtorhinus lividipennis Reuter is an important predator that preys on rice planthopper eggs and young nymphs, as a primary food source. Alanine aminotransferase (ALT) acts in many physiological and biochemical processes in insects. We cloned the full-length complementary DNA of C. lividipennis ClALT. Expression analysis showed higher expression in the fat body and midgut compared to other tissues. It is expressed in all C. lividipennis developmental stages and at least four organs. Silencing of ClALT by RNA interference significantly decreased the ClALT enzyme activity and ClALT expression compared to dsGFP-treated controls at 2 days after emergence (DAE). Silencing of ClALT influenced free hemolymph amino acid compositions, resulting in a reduction of Aspartic acid (Asp) and Alanine (Ala) proportions, and increased Cysteine (Cys) and Valine (Val) proportions in females at 2 DAE. dsClALT treatments led to decreased soluble total protein concentrations in ovary and fat body, and to lower reduced vitellogenin (Vg) expression, body weight, and the numbers of laid eggs. The double-stranded RNA viruse treatments also led to prolonged preoviposition periods and hindered ovarian development. Western blot analysis indicated that silencing ClALT also led to reduced fat body Vg protein abundance at 2 DAE. These data support our hypothesis that ClALT influences amino acid metabolism and fecundity in C. lividipennis.


Assuntos
Alanina Transaminase , Aminoácidos/metabolismo , Fertilidade , Heterópteros , Alanina Transaminase/genética , Alanina Transaminase/metabolismo , Aminoácidos/genética , Animais , Hemolinfa/metabolismo , Heterópteros/genética , Heterópteros/metabolismo , Heterópteros/fisiologia , Proteínas de Insetos/metabolismo , Interferência de RNA , Vitelogeninas/metabolismo
6.
Commun Biol ; 4(1): 856, 2021 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-34244602

RESUMO

Mosquito vectors transmit various diseases through blood feeding, required for their egg development. Hence, blood feeding is a major physiological event in their life cycle, during which hundreds of genes are tightly regulated. Blood is a rich source of proteins for mosquitoes, but also contains many other molecules including microRNAs (miRNAs). Here, we found that human blood miRNAs are transported abundantly into the fat body tissue of Aedes aegypti, a key metabolic center in post-blood feeding reproductive events, where they target and regulate mosquito genes. Using an artificial diet spiked with the mimic of an abundant and stable human blood miRNA, hsa-miR-21-5p, and proteomics analysis, we found over 40 proteins showing differential expression in female Ae. aegypti mosquitoes after feeding. Of interest, we found that the miRNA positively regulates the vitellogenin gene, coding for a yolk protein produced in the mosquito fat body and then transported to the ovaries as a protein source for egg production. Inhibition of hsa-miR-21-5p followed by human blood feeding led to a statistically insignificant reduction in progeny production. The results provide another example of the involvement of small regulatory molecules in the interaction of taxonomically vastly different taxa.


Assuntos
Aedes/metabolismo , MicroRNAs/sangue , Mosquitos Vetores/metabolismo , Vitelogeninas/metabolismo , Aedes/citologia , Aedes/genética , Animais , Linhagem Celular , Cromatografia Líquida/métodos , Corpo Adiposo/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Proteínas de Insetos/metabolismo , MicroRNAs/genética , Mosquitos Vetores/genética , Proteômica/métodos , RNA-Seq/métodos , Espectrometria de Massas em Tandem/métodos , Vitelogeninas/genética
7.
Sci Total Environ ; 791: 147984, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34118657

RESUMO

Nanoplastics (≤100 nm) represent the smallest fraction of plastic litter and may result in the aquatic environment as degradation products of larger plastic material. To date, few studies focused on the interactions of micro- and nanoplastics with freshwater Decapoda. The red swamp crayfish (Procambarus clarkii, Girard, 1852) is an invasive species able to tolerate highly perturbed environments. As a benthic opportunistic feeder, this species may be susceptible to plastic ingestion. In this study, adult P. clarkii, at intermolt stage, were exposed to 100 µg of 100 nm carboxylated polystyrene nanoparticles (PS NPs) through diet in a 72 h acute toxicity test. An integrated approach was conceived to assess the biological effects of PS NPs, by analyzing both transcriptomic and physiological responses. Total hemocyte counts, basal and total phenoloxidase activities, glycemia and total protein concentration were investigated in crayfish hemolymph at 0 h, 24 h, 48 h and 72 h from PS NPs administration to evaluate general stress response over time. Differentially expressed genes (DEGs) in the hemocytes and hepatopancreas were analyzed to ascertain the response of crayfish to PS NP challenge after 72 h. At a physiological level, crayfish were able to compensate for the induced stress, not exceeding generic stress thresholds. The RNA-Sequencing analysis revealed the altered expression of few genes involved in immune response, oxidative stress, gene transcription and translation, protein degradation, lipid metabolism, oxygen demand, and reproduction after PS NPs exposure. This study suggests that a low concentration of PS NPs may induce mild stress in crayfish, and sheds light on molecular pathways possibly involved in nanoplastic toxicity.


Assuntos
Astacoidea , Vitelogeninas , Animais , Hepatopâncreas/metabolismo , Estresse Oxidativo , Poliestirenos/metabolismo , Vitelogeninas/genética , Vitelogeninas/metabolismo
8.
Aquat Toxicol ; 236: 105843, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34010734

RESUMO

Octocrylene (OC) is a broad-spectrum ultraviolet-absorbing chemical used in sunscreen and other personal care products. Its health effects are a concern because it has been detected in water, fish, humans, and food chains. In vivo and in vitro investigations were performed in zebrafish (Danio rerio) larvae and a zebrafish liver cell line (ZFL), respectively, to understand the potential risks and molecular mechanisms of OC toxicity. The 96-h median lethal concentration (LC50) of OC was determined to be 251.8 µM in larvae and 5.5 µM in ZFL cells. Quantitative real-time PCR (qRT-PCR) showed that OC induced the expression of genes for CYPs (CYP1A, CYP3A65), estrogen receptors (ERα, ERß1, GPER), vitellogenin (VTG1), and sex determination (BRCA2, CYP19A, DMRT1, SOX9A), both in vitro and in vivo. A whole-transcriptome sequencing method was used to evaluate the gene expression profile of larvae exposed to OC. OC was found to mediate the biosynthesis of estrogens (such as estriol) and affect the antioxidant pathway (glutathione transferases and peroxisome). These findings clarify the toxic effects and molecular mechanisms of OC and support banning its use in cosmetics.


Assuntos
Acrilatos/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Linhagem Celular , Estrogênios/metabolismo , Hepatócitos/efeitos dos fármacos , Humanos , Larva/efeitos dos fármacos , Dose Letal Mediana , Fígado/efeitos dos fármacos , Protetores Solares/toxicidade , Transcriptoma , Vitelogeninas/metabolismo , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética
9.
Aquat Toxicol ; 236: 105868, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34051627

RESUMO

2,4-Dichlorophenol (2,4-DCP), an estrogenic endocrine disruptor, is widely spread in aquatic environments and may interfere with normal physiological functions in fish. However, the influence of this chemical on the synthesis of sex hormones is not well understood. In the present study, zebrafish (Danio rerio) were exposed to 2,4-DCP (80 and 160 µg/L) with or without fadrozole (an aromatase inhibitor which inhibits the synthesis of estradiol) from 20 to 40 days post fertilization. Then, the sex ratio, the content of vitellogenin (VTG) and sex hormones (androstenedione (ASD), estrone (E1), 17ß-estradiol (E2), estriol (E3), testosterone (T) and 11-ketotestosterone (11-KT)) were studied. Furthermore, the expression of genes involved in synthesis of sex hormones (cyp19a1a, cyp19a1b, 17ß-hsd, 11ß-hsd and cyp11b) along with the DNA methylation in cyp19a1a and cyp19a1b promoters was analyzed. The results showed that 2,4-DCP exposure led to female-biased ratio, increased the content of ASD, E2 and VTG, as well as the ratio of E2/11-KT, while decreased the levels of androgens (T and 11-KT). The sex hormonal change can be explained by the significant up-regulation of cyp19a1a, cyp19a1b, 17ß-hsd and 11ß-hsd genes. In addition, hypomethylation of cyp19a1a promoter was involved in this process. Notably, fadrozole can partly attenuate 2,4-DCP-induced feminization, and recover the levels of ASD, E2 and 11-KT. Thus, these results demonstrate that 2,4-DCP induces feminization in fish by disrupting the synthesis of sex hormones.


Assuntos
Clorofenóis/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Inibidores da Aromatase , Metilação de DNA/efeitos dos fármacos , Disruptores Endócrinos , Estradiol , Estrogênios/farmacologia , Fadrozol , Feminino , Feminização/genética , Hormônios Esteroides Gonadais , Humanos , Masculino , Fenóis , Razão de Masculinidade , Vitelogeninas/metabolismo , Peixe-Zebra/metabolismo
10.
Artigo em Inglês | MEDLINE | ID: mdl-33894369

RESUMO

Investigating the mechanisms of action of natural bioactive products against pests is a vital strategy to develop novel promising biopesticides. Scoparone, isolated from Artemisia capillaris, exhibited potent oviposition inhibition activity against Tetranychus cinnabarinus (a crop-threatening mite pests with strong fecundity). To explore the underlying mechanism, the vitellogenin (Vg) protein content, and Vg gene expression of mites from three consecutive generations of G0 individuals exposed to scoparone were determined, revealing marked inhibition. This study is the first to explore the egg development defect behaviour of mite pests induced by scoparone. The egg-laying inhibition of mites by scoparone was significantly increased by 47.43% compared with that of the control when TcVg was silenced by RNA interference (RNAi), suggesting that egg-development inhibition of female T. cinnabarinus by scoparone was mediated by low Vg gene expression. Furthermore, scoparone bound to the Vg protein in vitro, and its Kd value was 218.9 µM, implying its potential function in inhibiting the egg development of mites by directly targeting the Vg protein. This study will lay the foundation for the future applications of scoparone as an agrochemical for controlling the strong egg-laying capacity mite pests in agriculture.


Assuntos
Acaricidas/farmacologia , Cumarínicos/farmacologia , Óvulo/efeitos dos fármacos , Tetranychidae/efeitos dos fármacos , Animais , Clonagem Molecular , Regulação da Expressão Gênica/efeitos dos fármacos , Filogenia , Vasodilatadores/farmacologia , Vitelogeninas/metabolismo
11.
Sci Rep ; 11(1): 7326, 2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33795753

RESUMO

The cladoceran crustacean Daphnia has long been a model of energy allocation studies due to its important position in the trophic cascade of freshwater ecosystems. However, the loci for controlling energy allocation between life history traits still remain unknown. Here, we report CRISPR/Cas-mediated target mutagenesis of DNA methyltransferase 3.1 (DNMT3.1) that is upregulated in response to caloric restriction in Daphnia magna. The resulting biallelic mutant is viable and did not show any change in growth rate, reproduction, and longevity under nutrient rich conditions. In contrast, under starved conditions, the growth rate of this DNMT3.1 mutant was increased but its reproduction was reciprocally reduced compared to the wild type when the growth and reproduction activities competed during a period from instar 4 to 8. The life span of this mutant was significantly shorter than that of the wild type. We also compared transcriptomes between DNMT3.1 mutant and wild type under nutrient-rich and starved conditions. Consistent with the DNMT3.1 mutant phenotypes, the starved condition led to changes in the transcriptomes of the mutant including differential expression of vitellogenin genes. In addition, we found upregulation of the I am not dead yet (INDY) ortholog, which has been known to shorten the life span in Drosophila, explaining the shorter life span of the DNMT3.1 mutant. These results establish DNMT3.1 as a key regulator for life span and energy allocation between growth and reproduction during caloric restriction. Our findings reveal how energy allocation is implemented by selective expression of a DNMT3 ortholog that is widely distributed among animals. We also infer a previously unidentified adaptation of Daphnia that invests more energy for reproduction than growth under starved conditions.


Assuntos
DNA (Citosina-5-)-Metiltransferases/genética , Daphnia/metabolismo , Privação de Alimentos , Longevidade , Adaptação Fisiológica , Alelos , Animais , Tamanho Corporal , Sistemas CRISPR-Cas , Regulação da Expressão Gênica , Traços de História de Vida , Mitose , Biologia Molecular , Mutação , Fenótipo , RNA/metabolismo , RNA-Seq , Reprodução , Transcriptoma , Vitelogeninas/metabolismo
12.
Mar Drugs ; 19(4)2021 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-33806251

RESUMO

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val115, ASP116, Val117, and Lys122) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein-toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.


Assuntos
Proteínas de Peixes/metabolismo , Takifugu/metabolismo , Tetrodotoxina/metabolismo , Vitelogeninas/metabolismo , Fator de von Willebrand/metabolismo , Animais , Proteínas de Peixes/genética , Simulação de Acoplamento Molecular , Ligação Proteica , Domínios e Motivos de Interação entre Proteínas , Vitelogeninas/genética , Fator de von Willebrand/genética
13.
Aquat Toxicol ; 233: 105788, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33662878

RESUMO

The gene expression response thought to underlie the negative apical effects resulting from estrogen exposure have been thoroughly described in fish. Although epigenetics are believed to play a critical role translating environmental exposures into the development of adverse apical effects, they remain poorly characterized in fish species. This study investigated alterations of DNA methylation of estrogen receptor alpha (esr1) in brain and liver tissues from 8 to 10 month old male fathead minnows (Pimephales promelas) after a 2d exposure to either 2.5 ng/L or 10 ng/L 17α-ethynylestradiol (EE2). Changes in the patterns of methylation were evaluated using targeted deep sequencing of bisulfite treated DNA in the 5' region of esr1. Methylation and gene expression were assessed at 2d of exposure and after a 7 and 14d depuration period. After 2d EE2 exposure, males exhibited significant demethylation in the 5' upstream region of esr1 in liver tissue, which was inversely correlated to gene expression. This methylation pattern reflected what was seen in females. No gene body methylation (GBM) was observed for liver of exposed males. Differential methylation was observed for a single upstream CpG site in the liver after the 14d depuration. A less pronounced methylation response was observed in the upstream region in brain tissue, however, several CpGs were necessarily excluded from the analysis. In contrast to the liver, a significant GBM response was observed across the entire gene body, which was sustained until at least 7d post-exposure. No differential expression was observed in the brain, limiting functional interpretation of methylation changes. The identification of EE2-dependent changes in methylation levels strongly suggests the importance of epigenetic mechanisms as a mediator of the organismal response to environmental exposures and the need for further characterization of the epigenome. Further, differential methylation following depuration indicates estrogenic effects persist well after the active exposure, which has implications for the risk posed by repeated exposures..


Assuntos
Cyprinidae/metabolismo , Metilação de DNA/efeitos dos fármacos , Receptor alfa de Estrogênio/genética , Etinilestradiol/toxicidade , Expressão Gênica/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Cyprinidae/genética , Estrogênios/metabolismo , Feminino , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Vitelogeninas/metabolismo
14.
Fish Physiol Biochem ; 47(4): 797-810, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33665751

RESUMO

In the last decades, oestrogenic compounds have often been reported in environmentally relevant concentrations in aquatic environments around the world. Most laboratory studies of oestrogens try to understand the effects of a single contaminant, but in natural environments, the effects may be quite different due to interactions with other compounds. The present study aimed to compare the action of oestrone (E1) and bisphenol-A (BPA), acting singularly and in combination, on the spermatogenesis of Astyanax bimaculatus. After exposure to 100 ng/L of E1, BPA and a mixture of the two for 15 days, our results showed that E1 and the E1 + BPA mixture significantly altered the number of spermatogenic cells. BPA presented high cytotoxicity when compared to other treatments. Analysis of the two oestrogenic compounds suggests that the E1 + BPA mixture has no additive or synergistic effects. Together, the results of the present study indicate that endocrine-disrupting chemicals (EDCs) analysed alone may behave differently than when administered with other substances.


Assuntos
Compostos Benzidrílicos/toxicidade , Characidae , Disruptores Endócrinos/toxicidade , Estrogênios/toxicidade , Estrona/toxicidade , Fenóis/toxicidade , Poluentes Químicos da Água/toxicidade , Animais , Apoptose/efeitos dos fármacos , Characidae/metabolismo , Estradiol/metabolismo , Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Reprodução/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Testículo/efeitos dos fármacos , Testículo/patologia , Vitelogeninas/metabolismo
15.
Arch Insect Biochem Physiol ; 106(3): e21775, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33644918

RESUMO

DDX3 represents a well-defined subfamily of DEAD-box RNA helicase and exerts multiple functions in RNA metabolism, cell cycle, tumorigenesis, signal pathway, and fertility. Our previous study has shown that LmDDX3, the ortholog of DDX3 in Locusta migratoria, is ubiquitously expressed, and with a high abundance in testis and ovary. Knockdown of LmDDX3 results in a lethal phenotype in nymph, but it still remains unclear for its role in reproductive process. In this study, we therefore characterized LmDDX3 expression in female adult locust and analyzed its function in oocyte development. LmDDX3 was expressed in all tissues examined with significant more transcripts in ovary and hindgut. In ovary, a strong expression level was detected at the day just after adult eclosion, and a dramatic reduction then occurred during the oocyte development. LmDDX3 RNAi led to a reduced vitellogenin (Vg) expression in fat body via partially at least, the JH signaling pathway, and caused an upregulation of vitellogenin receptor (VgR) in ovary, and thus blocked the ovarian development and oocyte maturation. Sequence and phylogenetic analysis indicated that LmDDX3 was closely related to termite DDX3. Taken together, these data reveal a critical role for LmDDX3 in regulating the transcription of Vg and VgR, two major factors in vitellogenesis that is a key process required for ovary development and oocyte maturation in locust, and contribute thereof a new putative target for locust biological control.


Assuntos
Locusta migratoria , Oócitos/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , RNA Helicases , Animais , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Feminino , Expressão Gênica , Genes de Insetos , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Hormônios Juvenis/metabolismo , Locusta migratoria/genética , Locusta migratoria/fisiologia , Ninfa/genética , Ninfa/fisiologia , Oogênese/fisiologia , Ovário/metabolismo , Filogenia , RNA Helicases/genética , RNA Helicases/metabolismo , Interferência de RNA , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Transdução de Sinais , Vitelogênese/fisiologia , Vitelogeninas/genética , Vitelogeninas/metabolismo
16.
Artigo em Inglês | MEDLINE | ID: mdl-33540188

RESUMO

This study was done to evaluate the effects of Bisphenol A (BPA) on Siberian sturgeon (Acipenser baerii). As liver is the main organ in the homeostatic adjustments to stress, we used a proteomics method to address molecular response in this tissue. Also, we compared the levels of vitellogenin in plasma and mucus to propose that the last one be a non-invasive method to analyze this biomarker. The fish received 1, 10, and 100 µg g-1 week-1 BPA intraperitoneally for two weeks. The samples were taken on days 0, 7, and 14. Plasma vitellogenin level increased as the highest value was recorded in the group with 100 µg g-1 week-1 of BPA. Changes in the mucus and blood vitellogenin showed a similar pattern, suggesting that mucus could be used for evaluating the changes in blood vitellogenin. Comparative proteomics was used to determine the proteome of the liver of A. baerii in the highest dose of BPA in comparison with the control. Sixteen proteins were identified that their expression changed at least twice between the studied groups. The proteomic results showed that BPA increased the expression of proteins involved in the detoxification and metabolism, activated glycolysis, and produced necrosis in the liver.


Assuntos
Compostos Benzidrílicos/efeitos adversos , Disruptores Endócrinos/efeitos adversos , Poluentes Ambientais/efeitos adversos , Proteínas de Peixes/metabolismo , Peixes/metabolismo , Fenóis/efeitos adversos , Vitelogeninas/metabolismo , Animais , Proteínas de Peixes/análise , Proteínas de Peixes/sangue , Peixes/sangue , Fígado/efeitos dos fármacos , Fígado/metabolismo , Proteoma/análise , Proteoma/metabolismo , Vitelogeninas/análise , Vitelogeninas/sangue
17.
PLoS One ; 16(2): e0245928, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33571307

RESUMO

Vitellogenins, major yolk protein precursors, play an essential role in the reproduction and spread of all oviparous species, including insects. To investigate reproductive strategies of the warehouse moth Cadra cautella at the molecular level, a partial transcript of the C. cautella vitellogenin (CcVg) gene was extended through the rapid amplification of cDNA ends PCR and sequenced. The complete CcVg mRNA transcript was 5,334 bp long, which encoded a protein of 1,778 amino acids, including the first 14 amino acids of the signal peptide. The deduced CcVg protein contained a putative cleavage site (RTRR) at the amino-terminal side, similar to several other insect species. DGQR and GI/LCG motifs were present at the CcVg gene C-terminus, followed by nine cysteine residues. CcVg harbored 131 putative phosphorylation sites, numbering 84, 19, and 28 sites for serine, threonine, and tyrosine, respectively. The transcript showed a great resemblance with other lepidopteran Vgs. CcVg protein analysis revealed three conserved regions: 1) vitellogenin-N domain, 2) DUF 1943 (domain of unknown function), and 3) a von Willebrand factor type D domain. Additionally, sex, stage-specific, and developmental expression profiles of the CcVg gene were determined through RT-PCR. The Vg was first expressed in 22-day-old female larvae, and its expression increased with growth. The phylogenetic analysis based on different insect Vgs revealed that the CcVg exhibited close ancestry with lepidopterans. The CcVg-based RNAi experiments were performed, and the effects were critically evaluated. The qRT-PCR results showed that CcVg-based dsRNA suppressed the Vg gene expression up to 90% at 48 h post-injection. Moreover, CcVg-based RNAi effects resulted in low fecundity and egg hatchability in the CcVg-based dsRNA-treated females. The females laid eggs, but because of insufficient yolk protein availability the eggs could not succeed to hatch. The significant difference in the fecundity and hatchability unveils the importance of CcVg gene silencing and confirmed that the Vg gene plays a key role in C. cautella reproduction and it has the potential to be used as a target for RNAi-mediated control of this warehouse pest.


Assuntos
Mariposas/genética , Mariposas/fisiologia , Oogênese/genética , Interferência de RNA , Vitelogeninas/deficiência , Vitelogeninas/genética , Sequência de Aminoácidos , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Mariposas/crescimento & desenvolvimento , Caracteres Sexuais , Vitelogeninas/química , Vitelogeninas/metabolismo
18.
Insect Mol Biol ; 30(3): 277-286, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33427366

RESUMO

Social insects are notable for having two female castes that exhibit extreme differences in their reproductive capacity. The molecular basis of these differences is largely unknown. Vitellogenin (Vg) is a powerful antioxidant and insulin-signalling regulator used in oocyte development. Here we investigate how Royal Jelly (the major food of honeybee queens) and queen mandibular pheromone (a major regulator of worker fertility), affect the longevity and reproductive status of honey bee workers, the expression of Vg, its receptor VgR and associated regulatory proteins. We find that Vg is expressed in the ovaries of workers and that workers fed a queen diet of Royal Jelly have increased Vg expression in the ovaries. Surprisingly, we find that expression of Vg is not associated with ovary activation in workers, suggesting that this gene has potentially acquired non-reproductive functions. Therefore, Vg expression in the ovaries of honeybee workers provides further support for the Ovarian Ground Plan Hypothesis, which argues that genes implicated in the regulation of reproduction have been co-opted to regulate behavioural differences between queens and workers.


Assuntos
Abelhas/fisiologia , Evolução Biológica , Expressão Gênica , Proteínas de Insetos/genética , Traços de História de Vida , Vitelogeninas/genética , Animais , Abelhas/genética , Feminino , Proteínas de Insetos/metabolismo , Ovário/metabolismo , Reprodução/genética , Comportamento Social , Vitelogeninas/metabolismo
19.
Insect Biochem Mol Biol ; 130: 103526, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33453353

RESUMO

Insulins are peptide hormones widely studied for their important regulatory roles in metabolism, growth and development. In insects, insulin signaling along with the target of rapamycin (ToR) are involved in detecting and interpreting nutrient levels. Recently, by transcriptome analysis we reported an up-regulation of transcripts involved in insulin/ToR signaling in unfed Rhodnius prolixus; however, this signaling pathway is only activated in fed insects. Here, continuing with the blood-gorging triatomine R. prolixus as a model, we report the direct effect of insulin/ToR signaling on reproductive performance. By immunofluorescence we identified cells in the brain with positive signal to the R. prolixus ILP (Rhopr-ILP1) and show that the insulin receptor and protein effectors downstream of insulin/ToR signaling activation, are differentially expressed in ovarian follicles dependent on their developmental stage. Using qPCR we find that the expression of transcripts involved in insulin signaling in the central nervous system (CNS), fat body and ovaries increase as the state of starvation progresses, promoting a more highly sensitized state to respond rapidly to ILP/IGF levels. In addition, using dsRNA injection and in vivo and ex vivo assays to promote signaling activation we demonstrate a direct participation of insulin/ToR signaling in coordinating the synthesis of the main yolk protein precursor, vitellogenin, thereby influencing the numbers of eggs laid per female. We thereby show a mechanism by which nutritional signaling regulates reproductive performance in a vector of Chagas disease. As reproduction is responsible for propagation of insect populations, this work is important for the development of innovative biocontrol methods.


Assuntos
Insulina/metabolismo , Rhodnius , Serina-Treonina Quinases TOR/metabolismo , Vitelogeninas/metabolismo , Animais , Doença de Chagas/transmissão , Hormônios de Inseto/metabolismo , Insetos Vetores/genética , Insetos Vetores/metabolismo , Insetos Vetores/fisiologia , Receptor de Insulina/metabolismo , Reprodução/fisiologia , Rhodnius/genética , Rhodnius/metabolismo , Rhodnius/fisiologia , Transdução de Sinais
20.
Sci Rep ; 11(1): 2194, 2021 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-33500499

RESUMO

The sea cucumber Holothuria scabra is an economically valuable marine species which is distributed throughout the Asia-Pacific region. With the natural population declining due to over fishing, aquaculture of this species is deemed necessary. Hence, it is essential to understand the mechanisms regulating the reproduction in order to increase their populations. Sex steroids, including estrogens, androgens and progestogens, play an important role in reproduction in most vertebrates and several invertebrates. It has been proposed that sea cucumbers have the same sex steroids as vertebrates but the steroidogenic pathway in the sea cucumbers is still unclear. In this study, we demonstrated by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) that sex steroids (estradiol, progesterone, and testosterone) were present in H. scabra neural and gonadal tissues. In silico searches of available sea cucumber transcriptome data identified 26 steroidogenesis-related genes. Comparative analysis of encoded proteins for the steroidogenic acute regulatory protein (HscStAR), CYP P450 10, 17 and 3A (HscCYP10, HscCYP17, HscCYP3A) and hydroxysteroid dehydrogenases (Hsc3ß-HSD, Hsc17ß-HSD) with other species was performed to confirm their evolutionary conservation. Gene expression analyses revealed widespread tissue expression. Real-time PCR analysis revealed that HscStAR, HscCYP10, Hsc3ß-HSD, and Hsc17ß-HSD gene expressions were similar to those in ovaries and testes, which increased during the gonad maturation. HscCYP17 mRNA was increased during ovarian development and its expression declined at late stages in females but continued high level in males. The expression of the HscCYP3A was high at the early stages of ovarian development, but not at other later stages in ovaries, however it remained low in testes. Moreover, a role for steroids in reproduction was confirmed following the effect of sex steroids on vitellogenin (Vtg) expression in ovary explant culture, showing upregulation of Vtg level. Collectively, this study has confirmed the existence of steroids in an echinoderm, as well as characterizing key genes associated with the steroidogenic pathway. We propose that sex steroids might also be associated with the reproduction of H. scabra, and the identification of biosynthetic genes enables future functional studies to be performed.


Assuntos
Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Pepinos-do-Mar/genética , Maturidade Sexual/genética , Esteroides/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Estradiol/farmacologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Gônadas/efeitos dos fármacos , Anotação de Sequência Molecular , Especificidade de Órgãos/efeitos dos fármacos , Especificidade de Órgãos/genética , Filogenia , Progesterona/farmacologia , Pepinos-do-Mar/efeitos dos fármacos , Maturidade Sexual/efeitos dos fármacos , Esteroides/metabolismo , Fatores de Tempo , Distribuição Tecidual/efeitos dos fármacos , Vitelogeninas/genética , Vitelogeninas/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...