Mepanipyrim and cyprodinil induce divergent temporal patterns of AhR-mediated responses in zebrafish (Danio rerio) embryos and larvae

Abstract Mepanipyrim and cyprodinil are widely used to control and/or prevent fungal diseases in fruit culture. They are widely detected in aquatic environment and numerous food commodities including fruit and fruit products. Different from TCDD, mepanipyrim and cyprodinil are more easily degraded and metabolized in the environment. However, the in vivo analysis of their metabolic dynamics is unclear and need to be further confirmed. In this study, zebrafish embryos were constantly exposed to 100 µg/L mepanipyrim or cyprodinil for 7 days. The temporal pattern of CYP1A and AhR2 expression and EROD enzyme activity at different time frames during embryonic and larval development of zebrafish were investigated. Our results showed that mepanipyrim and cyprodinil tend to accumulate in zebrafish during early embryonic developmental stages. Meanwhile, mepanipyrim and cyprodinil exposure could increase the expression level of cyp1a and ahr2 genes and EROD activity by a dynamic pattern in different developmental stages of zebrafish. Besides, their metabolites, which may accumulate in the zebrafish larvae, have strong AhR agonistic activity and showed strong AhR binding ability. Importantly, the risk of exposure to pesticides in embryo stage is huge, and should be paid more attention.

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Aflatoxins biosynthesis, toxicity and intervention strategies: A review

10.31221/osf.io/b95h2 ◽

2019 ◽

Author(s):

Chem Int

Keyword(s):

Developing Countries ◽

Molecular Basis ◽

Genetic Regulation ◽

Intervention Strategies ◽

Historical Background ◽

Economic Losses ◽

Factors Affecting ◽

Biomarkers Of Exposure ◽

Food Commodities

Aflatoxins (AFTs) are toxic products of fungal metabolism, associated with serious health consequences and substantial economic losses to agriculture, livestock and poultry sectors, particularly in the developing countries. This review outlines the current information on AFTs in terms of historical background, classification, relative occurrence and co-existence with other mycotoxins in various food commodities. The phenomenon of aflatoxin (AFT) biosynthesis has been elucidated with reference to molecular basis, genetic regulation and factors affecting the AFT production. Moreover, the in vivo disposition kinetics, toxicological action and toxico-pathological consequences of AFTs have also been highlighted. Currently employed strategies for the detection and detoxification of AFTs, biomarkers of exposure assessment, potential economic impact and regulatory considerations regarding the AFTs have been emphasized.

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Genome-Wide Transcriptomic Identification and Functional Insight of Lily WRKY Genes Responding to Botrytis Fungal Disease

Plants ◽

10.3390/plants10040776 ◽

2021 ◽

Vol 10 (4) ◽

pp. 776

Author(s):

Shipra Kumari ◽

Bashistha Kumar Kanth ◽

Ju young Ahn ◽

Jong Hwa Kim ◽

Geung-Joo Lee

Keyword(s):

Abiotic Stress ◽

Biotic Stress ◽

Developmental Stages ◽

Expression Patterns ◽

Lilium Longiflorum ◽

Biotic And Abiotic Stress ◽

Biotic Stresses ◽

Genome Wide

Genome-wide transcriptome analysis using RNA-Seq of Lilium longiflorum revealed valuable genes responding to biotic stresses. WRKY transcription factors are regulatory proteins playing essential roles in defense processes under environmental stresses, causing considerable losses in flower quality and production. Thirty-eight WRKY genes were identified from the transcriptomic profile from lily genotypes, exhibiting leaf blight caused by Botrytis elliptica. Lily WRKYs have a highly conserved motif, WRKYGQK, with a common variant, WRKYGKK. Phylogeny of LlWRKYs with homologous genes from other representative plant species classified them into three groups- I, II, and III consisting of seven, 22, and nine genes, respectively. Base on functional annotation, 22 LlWRKY genes were associated with biotic stress, nine with abiotic stress, and seven with others. Sixteen unique LlWRKY were studied to investigate responses to stress conditions using gene expression under biotic and abiotic stress treatments. Five genes—LlWRKY3, LlWRKY4, LlWRKY5, LlWRKY10, and LlWRKY12—were substantially upregulated, proving to be biotic stress-responsive genes in vivo and in vitro conditions. Moreover, the expression patterns of LlWRKY genes varied in response to drought, heat, cold, and different developmental stages or tissues. Overall, our study provides structural and molecular insights into LlWRKY genes for use in the genetic engineering in Lilium against Botrytis disease.

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Zika Virus Infection Leads to Demyelination and Axonal Injury in Mature CNS Cultures

Viruses ◽

10.3390/v13010091 ◽

2021 ◽

Vol 13 (1) ◽

pp. 91

Author(s):

Verena Schultz ◽

Stephanie L. Cumberworth ◽

Quan Gu ◽

Natasha Johnson ◽

Claire L. Donald ◽

...

Keyword(s):

Nervous System ◽

Neural Development ◽

Zika Virus ◽

Developmental Stages ◽

Cell Types ◽

Neural Cells ◽

Zikv Infection ◽

Axonal Pathology ◽

Time Frames

Understanding how Zika virus (Flaviviridae; ZIKV) affects neural cells is paramount in comprehending pathologies associated with infection. Whilst the effects of ZIKV in neural development are well documented, impact on the adult nervous system remains obscure. Here, we investigated the effects of ZIKV infection in established mature myelinated central nervous system (CNS) cultures. Infection incurred damage to myelinated fibers, with ZIKV-positive cells appearing when myelin damage was first detected as well as axonal pathology, suggesting the latter was a consequence of oligodendroglia infection. Transcriptome analysis revealed host factors that were upregulated during ZIKV infection. One such factor, CCL5, was validated in vitro as inhibiting myelination. Transferred UV-inactivated media from infected cultures did not damage myelin and axons, suggesting that viral replication is necessary to induce the observed effects. These data show that ZIKV infection affects CNS cells even after myelination—which is critical for saltatory conduction and neuronal function—has taken place. Understanding the targets of this virus across developmental stages including the mature CNS, and the subsequent effects of infection of cell types, is necessary to understand effective time frames for therapeutic intervention.

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Developmental stages influence in vivo antimalarial activity of aerial part extracts of Schkuhria pinnata

Journal of Ethnopharmacology ◽

10.1016/j.jep.2021.114341 ◽

2021 ◽

pp. 114341

Author(s):

Catherine Nuwagira ◽

Emanuel L. Peter ◽

Clement Olusoji Ajayi ◽

John Adriko ◽

Kagoro-Rugunda Grace ◽

...

Keyword(s):

Aerial Part ◽

Developmental Stages ◽

Antimalarial Activity

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NF-κB activity during pancreas development regulates adult β-cell mass by modulating neonatal β-cell proliferation and apoptosis

Cell Death Discovery ◽

10.1038/s41420-020-00386-9 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Dror Sever ◽

Anat Hershko-Moshe ◽

Rohit Srivastava ◽

Roy Eldor ◽

Daniel Hibsher ◽

...

Keyword(s):

Cell Proliferation ◽

Developmental Stages ◽

Cell Mass ◽

Diabetes Incidence ◽

Β Cells ◽

Β Cell ◽

Cell Dysfunction ◽

Regulatory Circuit ◽

Proliferation And Apoptosis

AbstractNF-κB is a well-characterized transcription factor, widely known for its roles in inflammation and immune responses, as well as in control of cell division and apoptosis. However, its function in β-cells is still being debated, as it appears to depend on the timing and kinetics of its activation. To elucidate the temporal role of NF-κB in vivo, we have generated two transgenic mouse models, the ToIβ and NOD/ToIβ mice, in which NF-κB activation is specifically and conditionally inhibited in β-cells. In this study, we present a novel function of the canonical NF-κB pathway during murine islet β-cell development. Interestingly, inhibiting the NF-κB pathway in β-cells during embryogenesis, but not after birth, in both ToIβ and NOD/ToIβ mice, increased β-cell turnover, ultimately resulting in a reduced β-cell mass. On the NOD background, this was associated with a marked increase in insulitis and diabetes incidence. While a robust nuclear immunoreactivity of the NF-κB p65-subunit was found in neonatal β-cells, significant activation was not detected in β-cells of either adult NOD/ToIβ mice or in the pancreata of recently diagnosed adult T1D patients. Moreover, in NOD/ToIβ mice, inhibiting NF-κB post-weaning had no effect on the development of diabetes or β-cell dysfunction. In conclusion, our data point to NF-κB as an important component of the physiological regulatory circuit that controls the balance of β-cell proliferation and apoptosis in the early developmental stages of insulin-producing cells, thus modulating β-cell mass and the development of diabetes in the mouse model of T1D.

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Developmental stages, incubation temperature, and in vivo traceability of primordial germ cell in an important aquaculture species Piaractus mesopotamicus

Aquaculture ◽

10.1016/j.aquaculture.2021.736381 ◽

2021 ◽

Vol 535 ◽

pp. 736381

Author(s):

Geovanna Carla Zacheo Coelho ◽

Dilberto Ribeiro Arashiro ◽

Tamiris Disselli ◽

Matheus Pereira-Santos ◽

Tatiana María Mira-López ◽

...

Keyword(s):

Germ Cell ◽

Developmental Stages ◽

Incubation Temperature ◽

Primordial Germ Cell ◽

Piaractus Mesopotamicus

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Morpholino-Mediated Knockdown of ERα, ERβa, and ERβb mRNAs in Zebrafish (Danio rerio) Embryos Reveals Differential Regulation of Estrogen-Inducible Genes

Endocrinology ◽

10.1210/en.2013-1446 ◽

2013 ◽

Vol 154 (11) ◽

pp. 4158-4169 ◽

Cited By ~ 42

Author(s):

Lucinda B. Griffin ◽

Kathleen E. January ◽

Karen W. Ho ◽

Kellie A. Cotter ◽

Gloria V. Callard

Keyword(s):

Danio Rerio ◽

Specific Gene ◽

Estrogen Action ◽

Cytochrome P450 Aromatase ◽

Retained Intron ◽

Inducible Genes ◽

Hepatic Genes ◽

Danio Rerio Embryos

Genetically distinct estrogen receptor (ER) subtypes (ERα and ERβ) play a major role in mediating estrogen actions in vertebrates, but their unique and overlapping functions are not entirely clear. Although mammals have 1 gene of each subtype (ESR1 and ESR2), teleost fish have a single esr1 (ERα) and 2 esr2 (ERβa and ERβb) genes. To determine the in vivo role of different ER isoforms in regulating estrogen-inducible transcription targets, zebrafish (Danio rerio) embryos were microinjected with esr-specific morpholino (MO) oligonucleotides to disrupt splicing of the exon III/intron III junction in the DNA-binding domain. Each MO knocked down its respective normal transcript and increased production of variants with a retained intron III (esr1 MO) or a deleted or mis-spliced exon III (esr2a and esr2b MOs). Both esr1 and esr2b MOs blocked estradiol induction of vitellogenin and ERα mRNAs, predominant hepatic genes, but esr2b was the only MO that blocked induction of cytochrome P450 aromatase B mRNA, a predominant brain gene. Knockdown of ERβa with the esr2a MO had no effect on estrogen induction of the 3 mRNAs but, when coinjected with esr1 MO, attenuated the effect of ERα knockdown. Results indicate that ERα and ERβb, acting separately or cooperatively on specific gene targets, are positive transcriptional regulators of estrogen action, but the role of ERβa, if any, is unclear. We conclude that MO technology in zebrafish embryos is an advantageous approach for investigating the interplay of ER subtypes in a true physiological context.

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Intracellular distribution of α-galactosidase in leaves of Cucurbita pepo

Canadian Journal of Botany ◽

10.1139/b79-240 ◽

1979 ◽

Vol 57 (18) ◽

pp. 1904-1911 ◽

Cited By ~ 8

Author(s):

Brian Thomas ◽

John A. Webb

Keyword(s):

Cucurbita Pepo ◽

Developmental Stages ◽

Molecular Size ◽

Total Activity ◽

Intracellular Distribution ◽

Molecular Forms ◽

Enzyme Release ◽

Enzyme Preparations ◽

Isolated Protoplasts

The intracellular distribution of α-galactosidase in leaves of Cucurbita pepo was studied at different developmental stages using tissue strips, homogenates, and isolated protoplasts. About 85% of the total activity was found in the 500 g supernatant after tissues were homogenized either in water, in buffer at pH 5.6 or at pH 7.0, or in buffer containing 0.8 M KCl. Isolated protoplasts contained less than 10% of the total activity which was confined to the 20 000 g supernatant after lysis. p-Nitrophenyl-α-D-galactoside was readily hydrolysed when incubated with leaf strips but less than 3% of α-galactosidase could be leached from strips held for 4 h in 100 mM phosophate buffer or in buffer containing either 0.8 M KCl, 1 mM EDTA, or 1 mM dithioerythritol. It is concluded that at all stages of leaf development a high proportion of α-galactosidase is located in the exocellular region, not strongly bound either to the outer surface of the plasmalemma or to the cell wall but prevented from diffusing through the wall matrix by some physical attribute such as molecular size. Enzyme release occurred only following breakage or enzymatic digestion of the wall. The in vivo properties of the exocellular enzyme in leaf strips were compared with those of three molecular forms of α-galactosidase (LI, LII, and LIII) which were partially purified from mature leaves. The exocellular enzyme was active over a broad pH range with optima at pH 3.0 and pH 6.0; this resembles a combination of pH optima for LI and LIII. Inhibition by Cu2+ and p-chloromercuribenzoate resembled that for LIII and LII, respectively. Galactose and galactinol at a 5 mM concentration were 25–30% inhibitory for all enzyme preparations; melibiose, raffinose, and stachyose were very weakly inhibitory. The function of an exocellular α-galactosidase and its bearing on the transport of galactosylsucrose oligosaccharides to and from the minor veins of C. pepo are discussed.

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Aerobic and anaerobic metabolism for the zebrafish, Danio rerio, reared under normoxic and hypoxic conditions and exposed to acute hypoxia during development

Brazilian Journal of Biology ◽

10.1590/s1519-69842010000200027 ◽

2010 ◽

Vol 70 (2) ◽

pp. 425-434 ◽

Cited By ~ 40

Author(s):

WR Barrionuevo ◽

MN Fernandes ◽

O Rocha

Keyword(s):

Danio Rerio ◽

Anaerobic Metabolism ◽

Developmental Stages ◽

Acute Hypoxia ◽

Lactate Concentration ◽

Growth Delay ◽

Hypoxic Conditions ◽

Aerobic And Anaerobic ◽

Mild Hypoxia

In order to verify the influence of chronic and acute ambient oxygen levels from egg to adult stage of the zebrafish, in vivo oxygen consumption (MO2), critical tensions of oxygen (Pcrit), heart rate (fH) and total body lactate concentration (Lc) were determined for Danio rerio (Hamilton, 1822) raised at 28 °C under normoxic (7.5 mgO2.L-1 or 80 mm.Hg-1) and hypoxic conditions (4.3 mgO2.L-1) and exposed to acute hypoxia during different developmental stages. Our findings confirmed that very early stages do not respond effectively to ambient acute hypoxia. However, after the stage corresponding to the age of 30 days, D. rerio was able to respond to acute hypoxia through effective physiological mechanisms involving aerobic and anaerobic metabolism. Such responses were more efficient for the fishes reared under hypoxia which showed that D. rerio survival capability increased during acclimation to mild hypoxia. Measurements of body mass and length showed that moderate hypoxia did not affect growth significantly until the fish reached the stage of 60 days. Moreover, a growth delay was verified for the hypoxic-reared animals. Also, the D. rerio eggs-to-larvae survival varied from 87.7 to 62.4% in animals reared under normoxia and mild hypoxia, respectively. However, the surviving animals raised under moderated hypoxia showed a better aptitude to regulate aerobic and anaerobic capacities when exposed to acute hypoxia.

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Impaired Hair Follicle Morphogenesis and Polarized Keratinocyte Movement upon Conditional Inactivation of Integrin-linked Kinase in the Epidermis

Molecular Biology of the Cell ◽

10.1091/mbc.e07-06-0526 ◽

2008 ◽

Vol 19 (4) ◽

pp. 1462-1473 ◽

Cited By ~ 42

Author(s):

Kerry-Ann Nakrieko ◽

Ian Welch ◽

Holly Dupuis ◽

Dawn Bryce ◽

Agnieszka Pajak ◽

...

Keyword(s):

Hair Follicle ◽

Developmental Stages ◽

Single Cells ◽

Forward Movement ◽

Keratinocyte Proliferation ◽

Cultured Keratinocytes ◽

Conditional Inactivation ◽

Integrin Linked Kinase ◽

Key Aspects

Integrin-linked kinase (ILK) is key for cell survival, migration, and adhesion, but little is known about its role in epidermal development and homeostasis in vivo. We generated mice with conditional inactivation of the Ilk gene in squamous epithelia. These mice die perinatally and exhibit skin blistering and severe defects in hair follicle morphogenesis, including greatly reduced follicle numbers, failure to progress beyond very early developmental stages, and pronounced defects in follicular keratinocyte proliferation. ILK-deficient epidermis shows abnormalities in adhesion to the basement membrane and in differentiation. ILK-deficient cultured keratinocytes fail to attach and spread efficiently and exhibit multiple abnormalities in actin cytoskeletal organization. Ilk gene inactivation in cultured keratinocytes causes impaired ability to form stable lamellipodia, to directionally migrate, and to polarize. These defects are accompanied by abnormal distribution of active Cdc42 to cell protrusions, as well as reduced activation of Rac1 upon induction of cell migration in scraped keratinocyte monolayers. Significantly, alterations in cell spreading and forward movement in single cells can be rescued by expression of constitutively active Rac1 or RhoG. Our studies underscore a central and distinct role for ILK in hair follicle development and in polarized cell movements, two key aspects of epithelial morphogenesis and function.

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