Development of a Sensitive Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) for Vitellogenin in the Japanese Eel &lt;i&gt;Anguilla japonica&lt;/i&gt;

Okumura, Hitoshi; Hara, Akihiko; Saeki, Fumihiro; Todo, Takashi; Adachi, Souichi; Yamauchi, Kazuyo

doi:10.2331/fishsci.61.283

Cited by 34 publications

(19 citation statements)

References 2 publications

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“…The c-VTG ELISA, with a detection limit of 1 ng/ml, is one of the most sensitive ELISAs for VTG developed to date, and it compares very favorably with the most sensitive radioimmunoassays for VTG [24][25][26]. Tyler et al acteristics of the c-VTG ELISA are similar, with respect to inter-and intraassay variation, to other reported ELISAs for VTG [27][28][29][30]. Where assay sensitivity is not the main criteria, when measuring VTG in females, for example, the sensitivity of the assay can be reduced by modifying the incubation conditions (e.g., changing the concentration of either the coating VTG or the second antibody conjugate dilution) or by using an equilibrium assay rather than a nonequilibrium assay.…”

Section: Discussionmentioning

confidence: 86%

An in vivo testing system for endocrine disruptors in fish early life stages using induction of vitellogenin

Tyler¹,

Aerle²,

Hutchinson³

et al. 1999

Enviro Toxic and Chemistry

223

122

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An enzyme‐linked immunosorbent assay (ELISA) based oncarp (Cyprinus carpio) vitellogenin (c‐VTG, an estrogen‐dependent precursor of egg yolk) was developed to quantify VTG in the fathead minnow (Pimephales promelas) and was employed to establish an in vivo testing system for estrogens in fish early life stages. The working range of the c‐VTG ELISA was between 1 and 50 ng/ml (20%–80% binding). The inter‐ and intraassay variations were 9.7 ± 3% and 8.9 ± 3.4% (n = 15), respectively. Plasma and whole body homogenates from females and from estrogenized male fathead minnow diluted parallel with the c‐VTG standard. Measurements of plasma VTG fractionated on sodium dodecyl sulphate gels and quantified using densitometry were compared with measurements with the same plasma samples using the c‐VTG ELISA and they were similar for the carp and fathead minnow. The plasma concentration of VTG in the male fathead minnow was 25 ± 6 ng/ml, whereas in females, plasma concentrations of VTG ranged from 5.3 ± 1 μg/ml to 182 ± 7.6 μg/ml, depending on the state of maturity. Exposure of fathead minnows to estradiol‐17β in the water (nominal concentrations of 25 ng/L, 50 ng/L, and 100 ng/L) from 24 h postfertilization to 30 d posthatch induced VTG synthesis in a dose‐dependent manner. Shorter periods of exposure to 100 ng/L estradiol‐17β (24 h postfertilization to 10 or 20 d posthatch) similarly induced a vitellogenic response. This work demonstrates that fathead minnow are sensitive to estrogen and are able to synthesize VTG very early in development; this provides the basis for the adaptation of existing fish early‐life‐stage tests for the in vivo testing of estrogenic chemicals.

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Section: Discussionmentioning

confidence: 86%

An in vivo testing system for endocrine disruptors in fish early life stages using induction of vitellogenin

Tyler¹,

Aerle²,

Hutchinson³

et al. 1999

Enviro Toxic and Chemistry

223

122

View full text Add to dashboard Cite

show abstract

“…In plasma samples, these assays vary in their sensitivity, but some have the ability to detect very low levels of protein expression, i.e. 1 ng/ml or less [134-137]. Vtg assays based on polyclonal antibodies are generally restricted for use with the homologous species, but some antibodies do cross-react with Vtg in other species (e.g.…”

Section: Molecular Mechanisms For Oogenic Protein Gene Expressionmentioning

confidence: 99%

Eggshell and egg yolk proteins in fish: hepatic proteins for the next generation: oogenetic, population, and evolutionary implications of endocrine disruption

2003

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The oocyte is the starting point for a new generation. Most of the machinery for DNA and protein synthesis needed for the developing embryo is made autonomously by the fertilized oocyte. However, in fish and in many other oviparous vertebrates, the major constituents of the egg, i.e. yolk and eggshell proteins, are synthesized in the liver and transported to the oocyte for uptake. Vitellogenesis, the process of yolk protein (vitellogenin) synthesis, transport, and uptake into the oocyte, and zonagenesis, the synthesis of eggshell zona radiata proteins, their transport and deposition by the maturing oocyte, are important aspects of oogenesis. The many molecular events involved in these processes require tight, coordinated regulation that is under strict endocrine control, with the female sex steroid hormone estradiol-17β in a central role. The ability of many synthetic chemical compounds to mimic this estrogen can lead to unscheduled hepatic synthesis of vitellogenin and zona radiata proteins, with potentially detrimental effects to the adult, the egg, the developing embryo and, hence, to the recruitment to the fish population. This has led to the development of specific and sensitive assays for these proteins in fish, and the application of vitellogenin and zona radiata proteins as informative biomarkers for endocrine disrupting effects of chemicals and effluents using fish as test organisms. The genes encoding these important reproductive proteins are conserved in the animal kingdom and are products of several hundred million years of evolution.

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“…Sandwich ELISA was carried out according to the method of quantitating eel vitellogenin (Okumura et al, 1995) with some modifications as follows.…”

Section: Elisa Proceduresmentioning

confidence: 99%

Quantitative changes in yolk protein and other components in the ovary and testis of the sea urchin Pseudocentrotus depressus

Unuma

Yamamoto

Akiyama

et al. 2003

Journal of Experimental Biology

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SUMMARY Both male and female sea urchins accumulate the major yolk protein (MYP;the most abundant yolk granule protein in sea urchin eggs) in the nutritive phagocytes of immature gonads before gametogenesis. In this study,quantitative changes in MYP as well as in other biochemical components in the ovary and testis were examined in the course of gametogenesis in Pseudocentrotus depressus. Before gametogenesis, both the ovary and testis contained large quantities of proteins, lipids and polysaccharides. MYP reached about 80% of total protein in both sexes. In the testis, MYP decreased rapidly as spermatogenesis proceeded, and the fully mature testis contained little MYP; the levels of lipids and polysaccharides also decreased. In contrast, the levels of nucleic acids and proteins other than MYP increased markedly. In the ovary, MYP decreased gradually as oogenesis proceeded, and the fully mature ovary contained less than half of the initial amount of MYP. Polysaccharides also decreased, whereas proteins other than MYP increased. These results, taken together with those from other studies, suggest that MYP serves as a protein reserve that accumulates before gametogenesis and is used as material for synthesizing new substances constituting gametes in both male and female sea urchins.

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Development of a Sensitive Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) for Vitellogenin in the Japanese Eel <i>Anguilla japonica</i>

Cited by 34 publications

References 2 publications

An in vivo testing system for endocrine disruptors in fish early life stages using induction of vitellogenin

An in vivo testing system for endocrine disruptors in fish early life stages using induction of vitellogenin

Eggshell and egg yolk proteins in fish: hepatic proteins for the next generation: oogenetic, population, and evolutionary implications of endocrine disruption

Quantitative changes in yolk protein and other components in the ovary and testis of the sea urchin Pseudocentrotus depressus

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