Vtg Gene Research Articles

Genome-Wide Identification, Expression and Response to Estrogen of Vitellogenin Gene Family in Sichuan Bream (Sinibrama taeniatus).

To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6), type Ⅱ (vtg2) and type Ⅲ (vtg3) based on differences in their subdomain structure. The promoter sequence of vtgs has multiple estrogen response elements, and their abundance appears to correlate with the responsiveness of vtg gene expression to estrogen. Gene expression analyses revealed that the vitellogenesis of Sichuan bream involves both heterosynthesis and autosynthesis pathways, with the dominant pathway originating from the liver. The drug treatment experiments revealed that 17β-estradiol (E2) tightly regulated the level of vtg mRNA in the liver. Feeding fish with a diet containing 100 μg/g E2 for three weeks significantly induced vtg gene expression and ovarian development, leading to an earlier onset of vitellogenesis. Additionally, it was observed that the initiation of vtg transcription required E2 binding to its receptor, a process primarily mediated by estrogen receptor alpha in Sichuan bream. The findings of this study provide novel insights into the molecular information of the vitellogenin gene family in teleosts, thereby contributing to the regulation of gonadal development in farmed fish.

Genome-Wide Identification of Vitellogenin Gene Family and Comparative Analysis of Their Involvement in Ovarian Maturation in Exopalaemon carinicauda.

Vitellogenin (Vtg) is a precursor of yolk proteins in egg-laying vertebrates and invertebrates and plays an important role in vitellogenesis and embryonic development. However, the Vtg family remains poorly characterized in Exopalaemon carinicauda, a major commercial mariculture species found along the coasts of the Yellow and Bohai Seas. In this study, 10 Vtg genes from the genomes of E. carinicauda were identified and characterized. Phylogenetic analyses showed that the Vtg genes in crustaceans could be classified into four groups: Astacidea, Brachyra, Penaeidae, and Palaemonidae. EcVtg genes were unevenly distributed on the chromosomes of E. carinicauda, and a molecular evolutionary analysis showed that the EcVtg genes were primarily constrained by purifying selection during evolution. All putative EcVtg proteins were characterized by the presence of three conserved functional domains: a lipoprotein N-terminal domain (LPD_N), a domain of unknown function (DUF1943), and a von Willebrand factor type D domain (vWD). All EcVtg genes exhibited higher expression in the female hepatopancreas than in other tissues, and EcVtg gene expression during ovarian development suggested that the hepatopancreas is the main synthesis site in E. carinicauda. EcVtg1a, EcVtg2, and EcVtg3 play major roles in exogenous vitellogenesis, and EcVtg3 also plays a major role in endogenous vitellogenesis. Bilateral ablation of the eyestalk significantly upregulates EcVtg mRNA expression in the female hepatopancreas, indicating that the X-organ/sinus gland complex plays an important role in ovarian development, mostly by inducing Vtg synthesis. These results could improve our understanding of the function of multiple Vtg genes in crustaceans and aid future studies on the function of EcVtg genes during ovarian development in E. carinicauda.

Vitamin D3 promotes fish oocyte development by directly regulating gonadal steroid hormone synthesis†.

Vitamin D receptors and vitamin D3-metabolizing enzymes have been found to be highly expressed in the ovaries and spermatophores of fish. However, the role of vitamin D3 on fish gonadal development has rarely been reported. In this study, 2-month-old female zebrafish were fed with different concentrations of vitamin D3 diets (0, 700, 1400, and 11200IU/kg) to investigate the effects of vitamin D3 on ovarian development. The diet with 0IU/kg vitamin D3 resulted in elevated interstitial spaces, follicular atresia, and reproductive toxicity in zebrafish ovaries. Supplementation with 700 and 1400IU/kg of vitamin D3 significantly increased the oocyte maturation rate; upregulated ovarian gonadal steroid hormone synthesis capacity; and elevated plasma estradiol, testosterone, and ovarian vitellogenin levels. Furthermore, the current study identified a vitamin D response element in the cyp19a1a promoter and demonstrated that 1.25(OH)2D3-vitamin D response directly activated cyp19a1a production through activating the vitamin D response element. In conclusion, this study shows that an appropriate concentration of vitamin D3 can promote zebrafish ovarian development and affect vitellogenin synthesis through the vdr/cyp19a1a/er/vtg gene axis.

Influence of phthalate and non-phthalate plasticizers on reproductive endocrine system-related gene expression profiles in Japanese medaka (Oryzias latipes)

Plasticizers containing phthalates have the potential to alter endocrine function in vertebrates. While non-phthalate plasticizers were previously considered to be environmentally friendly and safe, our research team discovered that bis-(2-ethylhexyl) adipate (DEHA) and acetyl tributyl citrate (ATBC) disrupt thyroid hormones in Japanese medaka (Oryzias latipes). We assessed reproductive- and estrogen-responsive gene expression patterns in Japanese medaka to determine whether the phthalate plasticizers bis-(2-ethylhexyl) phthalate (DEHP, positive control) and the non-phthalate plasticizers DEHA and ATBC disrupt endocrine signaling. The results showed that the levels of choriogenin H (chgH) and vitellogenin (vtg) genes increased after exposure to DEHP and ATBC, suggesting that these plasticizers may have estrogenic activity. Exposure to DEHP and DEHA resulted in the upregulation of kisspeptin (kiss), gonadotropin-releasing hormone (gnrh), and follicle-stimulating hormone beta (fshβ) genes, suggesting that these plasticizers may interfere with reproductive function. To the best of our knowledge, this is the first study to demonstrate that the non-phthalate plasticizers DEHA and ATBC can disrupt reproduction-related hormonal activity in fish.

Single and mixture toxicity evaluation of avobenzone and homosalate to male zebrafish and H295R cells

Avobenzone and homosalate are widely used in sunscreens to provide ultraviolet (UV) protection, either as single compounds or in combination. Some UV filters exhibit estrogenic or anti-androgenic activities, however, studies regarding their interactions and toxicity in mixtures are limited. In this study, the effect of the toxicity of a binary mixture comprising avobenzone (0.72 μg L−1) and homosalate (1.02 and 103 μg L−1) on steroid hormone biosynthesis were investigated using male zebrafish and human adrenocortical carcinoma (H295R) cells. In fish exposed to homosalate, a significant decrease in the gonadosomatic index, testosterone level, and transcription of several genes (e.g, hsd3b2, cyp17a1, and hsd17b1) and a significant increase in the hepatosomatic index, liver steatosis, 17β-estradiol level, and transcription of vtg gene were observed. These results suggest that estrogenic and anti-androgenic effects of homosalate were mediated by the steroidogenic pathway. The presence of 0.72 μg L−1 of avobenzone augmented the anti-androgenic responses in male fish. The testosterone level in the H295R cells were significantly decreased after they were exposed to homosalate alone or in combination with avobenzone, which is consistent with observations in male zebrafish. Further studies need to be conducted to understand the endocrine disrupting properties of long-term exposure to substances typically used in sunscreens.

Expression Kinetics of Vitellogenin Gene in Estradiol-17β Induced <i>Catla catla</i>

Vitellogenin (Vtg) is the yolk precursor protein in fish and is synthesized in the liver under estradiol-17β (E2) stimulation. In all oviparous animals, Vtg is synthesized as a response to endogenous estrogen in females in the breeding season. It also could be induced exogenously by E2 in males and juveniles. In the current study the inducibility of Vtg gene in Indian major carp, Catla catla was tested. A semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay was developed for kinetic study of Vtg mRNA level in C. catla after primary and secondary inductions by E2. It was found that the level of Vtg-specific mRNA was highest following 12 h post treatment in primary induction, and after 1 d post treatment in secondary induction. There was a sharp decrease in Vtg mRNA level on day one and day two in primary and secondary induction, respectively. Thus the ‘memory effect’ of vitellogenin induction by E2 in catla could not be confirmed.

Endocrine disruptive toxicity of cypermethrin in Labeo catla: Involvement of genes and proteins related to the HPG axis

Cypermethrin (CYP) is a synthetic pyrethroid abundantly used in agriculture and aquaculture. It is an established potent endocrine disruptor to fish, yet the molecular mechanism behind its reproductive toxicity remains unclear. In this study, fish Labeo catla (Catla) was exposed to environmentally relevant concentration of CYP (0.7 μg/L) and 0.14 μg/L for 30 days. The changes in circulating sex steroids, genes, and hormones linked to the hypothalamic-pituitary-gonadal (HPG) axis, stress response and associated histological alterations were studied. Significant decline (P < 0.05) in serum 17 beta (β) estradiol (E2), 11 ketotestosterone (11-KT), and brain (FSH and GnRH) were observed in 0.7 μg/L dose of CYP. These effects may be due to the down-regulated expression of the upstream genes of the HPG axis i.e. Kiss 1 and Kiss 2, which further downregulates the expression of the GnRH gene. The decreased level of E2 and 11-KT also affects the vitellogenin (Vtg) gene expression, reducing the production of Vtg, a crucial protein for ovarian development. Principal component analysis (PCA) revealed the relationship between CYP and the biosynthesis of sex steroids. The toxic effect of CYP was also visible in antioxidant enzyme assay and related histological alterations. Overall, the study elucidated that long-term exposure to CYP, even at an environmentally relevant dose, may affect reproductive potential and fish recruitment. The study provides important insights into molecular mechanisms underlying CYP-induced endocrine disruption in fish, and it also raises questions about CYP's potential toxicity at environmentally relevant concentration in terms of understanding ecological risk.

Uma abordagem de multibiomarcadores para investigar os efeitos do paracetamol no eixo de regulação da reprodução de machos do bagre Neotropical Rhamdia quelen

Abstract: Aim Paracetamol (PCM), or acetaminophen, is one of the most used drugs for human treatment of pain and fever. Since it has often been found in the aquatic environment, the aim of this study was to investigate the effects of PCM on the reproductive axis of male Rhamdia quelen catfish. Methods Different biomarkers were evaluated in hypothalamus, liver and gonads, as well as the plasma sexual hormone quantification. The fish were exposed to three PCM concentrations: 0.25, 2.5 and 25 µg.L-1 and to a control group (solvent acetone 0.0003%). After 14 days of exposure, fish were anesthetized, for blood sampling and biometrics, and after euthanasia, the tissues were sampled. Plasma was used for 11- keto testosterone and 17β - estradiol quantification. The hypothalamus was collected for brain aromatase (cyp19a1b) gene expression; the liver for the vitellogenin (vtg) gene expression and biochemical biomarkers; and gonad for the biochemical and histological biomarkers analyses. Results No alterations were observed in the hormone’s levels, sexual maturation level or in cyp19a1b and vtg gene expression. In the liver the non-protein thiols concentration increased at 2.5 µg.L-1 of PCM, and the superoxide dismutase (SOD) activity was reduced at 0.25 µg.L-1 of PCM. In gonads, glutathione S-transferase (GST) activity decreased and SOD activity increased at 25 µg.L-1 of PCM, while the glutathione peroxidase (GPx) activity reduced after exposure to both PCM concentrations. Conclusion The results showed that environmental concentrations of PCM can cause alterations in the antioxidant system, mainly in the gonads of R. quelen males. However, without significant change in the hormones levels or in the expression of genes related to the reproduction axis. These alterations occurred at concentrations already found in aquatic environment, including in Brazil.

Evaluating the alterations of the estrogen-responsive genes in Cyprinodon variegatus larvae for biomonitoring the impacts of estrogenic endocrine disruptors (EEDs)

Currently, endocrine disruptors (EDs) can be found in all the environmental compartments. To understand the effects of estrogenic EDs (EEDs), adults of Cyprinodon variegatus have been classically used as a marine model. However, it is during development that exposure to contaminants may generate permanent consequences. Thus, the aim of this study was to verify the effects produced by acute exposure to 17α-ethinylestradiol (EE2) in C. variegatus larvae. Quantitative PCR (qPCR) results revealed the induction of vtg and zp gene expression on exposure to 1000 ng/L EE2 and the induction of vtgc, zp2, zp3 and cyp19a2, and inhibition of vtgab, wap and cyp1a1 on exposure to 100 ng/L EE2. Lower concentrations inhibited the gene expression of vtgab and wap (50 ng/L), cyp1a1 (25 ng/L) and zp2 (12.5 ng/L). These alterations in gene expression allow us to affirm that larvae of C. variegatus are an efficient and sensitive model for biomonitoring EEDs.

Effects of sex hormone rescue on gametogenesis in allotriploid crucian carp

Sex hormones play an important role in gametogenesis. During the breeding season, sex hormone levels in the sterile allotriploid crucian carp (3nAT) were significantly lower than those in the fertile diploid red crucian carp (2nRCC). Sex hormone rescue was performed to study the relationship between sex hormones and 3nAT sterility. For three months before breeding season, 3nAT females and males were fed the sex hormones estradiol and testosterone, respectively. During breeding season, hormone-rescued 3nAT fish (re-3nAT) showed normal reproductive behavior, and levels of estradiol in female re-3nAT and testosterone in male re-3nAT were similar to those of the diploid control (2nRCC). The vtg gene, which is associated with the hepatic synthesis of vitellogenin in the female, and the genes spef2, tekt1, and ift27, which are associated with the formation of the sperm flagellum, were significantly upregulated in re-3nAT as compared to 3nAT. Under artificially assisted reproduction, re-3nAT individuals produced a few eggs of different sizes and sperm with abnormal structures. However, few individuals had insemination ability. Neither zygotes produced by self-crossing re-3nAT nor those produced by crossing re-3nAT with the haploid gametes produced by 2nRCC exhibited normal embryonic development. Thus, although sex hormone treatment led to the production of abnormal gametes by re-3nAT, viable offspring did not develop, meaning that re-3nAT remained sterile. These results indicated that sex hormones affected gamete maturation but were not the determinant of triploid sterility.

Role of vitamin a in the ovary development for female Eriocheir sinensis in the gonadal development stage

This study aimed to test the regulatory role of vitamin A in the vitellogenesis of female Eriocheir sinensis. Three diets containing a gradient of vitamin A level (0-control, 8000 and 30,000 IU/kg) were fed to female crabs (74.44 ± 0.81 g) for 90 days. Compared to the vitamin A devoid group, 8000 IU/kg of dietary vitamin A increased the content of hemolymph ecdysone, vitellogenin levels and the gonadosomatic index. The 8000 IU/kg vitamin A level induced vitellogenin (vtg) gene expression in the ovary and hepatopancreas. The vtg gene expression was induced by upregulating retinoid x receptor, ecdysone receptor, ecdysteroid responsive gene E75 and vitellogenin receptor. Lipid accumulation was also improved for ovarian development. Hepatosomatic index and lipid accumulation in the hepatopancreas and ovary were increased by 8000 IU/kg vitamin A supplement relative to the control. The 8000 IU/kg vitamin A diet upregulated the gene expressions in lipid uptake, synthesis and export, but downregulated the genes related to catabolism in the hepatopancreas. Only genes related to lipid uptake were positively influenced by 8000 IU/kg of vitamin A in the ovary. Furthermore, proper vitamin A could improve the health of crabs by enhancing their antioxidant capacity and immunity. In contrast, a negative effect on hepatopancreas antioxidant ability, lipid metabolism and vitellogenesis was found due to an excess vitamin A supplement (30,000 IU/kg) compared to 8000 IU/kg of dietary vitamin A. This study indicates the necessity of vitamin A supplements in formulated diets because they can optimize lipid metabolism and promote vitellogenesis during ovarian development.

Regulation of vtg and VtgR in mud crab Scylla paramamosain by miR-34.

Vitellogenin (Vtg) is the precursor of major yolk protein and plays a crucial role in the maturation of oocytes and the production of eggs in oviparous animals. Vitellogenin receptor (VtgR) mediates the transport of Vtg explicitly to oocytes in the membrane. In a previous study, we found that miR-34 can regulate the expression of some eyestalk genes and affect reproduction in mud crab Scylla paramamosain, one of the most important economic crabs on the coasts of southern China. In this study, firstly, we found that miR-34 can target at 3'-UTR of Vtg and VtgR genes by using bioinformatic tools and predicted miR-34 might depress the expression of Vtg and VtgR. Secondly, the relative luciferase activity of HEK293T cells co-transfected with miRNA mimic and pmir-RB-REPORTTM-Vtg/VtgR-3'UTR was significantly lower than those of cells co-transfected with mimic NC and pmir-RB-REPORTTM-Vtg/VtgR-3'UTR. Finally, in vivo experiments showed that agomiR-34 could repress the expression of Vtg and VtgR genes, while Antigomir-34 could promote the expression of these two genes. These results confirm our hypothesis and previous published results that miR-34 may indirectly regulate ovarian development by binding to the 3'-UTR of Vtg and VtgR genes and inhibiting their expression.

Effects of polyvinyl chloride microplastics on reproduction, oxidative stress and reproduction and detoxification-related genes in Daphnia magna

The drastic increase of microplastics (MPs) in aquatic environment has become a serious threat to marine and freshwater ecosystems. However, little information is available regarding the potential detrimental effects of polyvinyl chloride microplastics (PVC-MPs) on aquatic organisms. This study investigated the changes of reproduction parameters, oxidative stress and the expression of reproduction and detoxification-related genes in Daphnia magna after exposed to 2 ± 1 and 50 ± 10 μm PVC-MPs. The results showed that chronic exposure to 2 ± 1 μm PVC-MPs prolonged days to the first brood, increased total number of broods per female and frequency of molting per adult, decreased offspring number at first brood and total number of offspring per female in D. magna. Moreover, 2 ± 1 μm PVC-MPs also disturbed the activities of SOD and CAT, increased GSH and MDA levels. The expression of Vtg, SOD, CAT, CYP314 and CYP360A8 genes also exhibited different response patterns depending on exposure time. Furthermore, 50 ± 10 μm PVC-MPs decreased offspring at first brood and Vtg mRNA level, increased the transcription levels and activities of SOD and CAT. These results suggest that the presence of PVC-MPs in aquatic environment may cause reproduction toxicity by disrupting the reproduction and detoxification-related genes expression and inducing oxidative stress in D. magna.

Tissue distribution and endocrine disruption effects of chronic exposure to pharmaceuticals and personal care products mixture at environmentally relevant concentrations in zebrafish.

Pharmaceuticals and personal care products (PPCPs) as emerging contaminants are ubiquitously present in the aquatic environment. Using in vivo and in silico techniques, this study aims to elucidate tissue distribution and endocrine disruption effects of chronic exposure (120 days) to PPCP mixture at environmentally relevant concentrations (ERCs) in adult zebrafish. Results from UHPLC-MS/MS analyses showed elevated distribution of PPCPs in zebrafish tissues in the order of liver > gonad > brain. Upregulation of steroid hormone receptors, both gonadotropin, and steroidogenic genes perturb the HPG axis pathway in females, while male fish exhibited significantly downregulated expressions of vtg, cyp17, and 17βhsd genes with inhibited fecundity. The Spearman correlation indicated a significant positive relationship between PPCPs bioaccumulation and mRNA levels of HPG axis genes. In silico molecular docking (MD) revealed specific amino acid residues of PPCPs binding with zebrafish estrogen receptors. Furthermore, the strongest binding energies of sulfamethoxazole, carbamazepine, and triclosan were discovered in erα and erβ estrogen receptors, confirming PPCPs' xenoestrogenic behavior. To summarize, chronic exposure to ERCs resulted in a high accumulation of PPCPs in the liver and gonad tissues of adult zebrafish, as well as associated perturbed genetic responses. As a result, strict environmental regulations for the disposal of PPCPs should be ensured to protect ecological and public health.

Effect of benzotriazole on oxidative stress response and transcriptional gene expression in Oryzias latipes and Danio rerio embryo

Emerging contaminants (EC) such as benzotriazole are being released into the environment in various ways, therefore it is necessary to understand how organisms are affected by EC. In this study, we exposed medaka (Oryzias latipes) and zebrafish (Danio rerio) during their embryonic period (1 day after hatching) to benzotriazole to investigate its effects on oxidative stress (ROS, GSH, GST, SOD, CAT and MDA) and changes in gene expression patterns. In both medaka and zebrafish, the influence of oxidative stress was confirmed through an increased MDA level and changes in the ROS and GSH levels. Antioxidant enzymes such as GST, CAT, and SOD were affected by benzotriazole; however, medaka and zebrafish showed different patterns in the effects by benzotriazole. Results of oxidative stress genes expression showed that medaka had either no influence or had a decrease in the gene expression profile, whereas zebrafish had a statistically significant increase in the expression of some genes. The cyp1a gene expression was increased in both species. However, vtg gene expression was increased only in zebrafish but decreased in medaka, indicating no estrogenic effects in medaka. Apoptosis genes showed changes in expression in both the species but was these changes were not dose-dependent. However, zebrafish caspase-9 gene expression was increased in all of the exposed groups, suggesting the effects on the intrinsic pathway associated with caspase-9. In conclusion, the results indicate that the toxic effects of benzotriazole differ at various levels in the two small fish medaka and zebrafish embryos.

The effect of polyethylene terephthalate and abamectin on oxidative damages and expression of vtg and cyp1a genes in juvenile zebrafish

Increasing the abundance and wide distribution of microplastics is one of the most important concerns of biological communities. These particles can act as carriers of environmental chemical pollutants and pesticides. Absorption of these particles by living organisms, especially aquatic organisms, may cause changes in the gene expression as well as enzymes activity in the antioxidant pathway. In this study, combined and individual effects of polyethylene terephthalate (PET) microplastics, as one of the most widely used plastics in industry, and abamectin (ABM) pesticide was evaluated on glutathione content, total antioxidant capacity, free radical scavenging, vitellogenin (vtg) and cytochrome P450, family 1, subfamily a (cyp1a) gene expression in zebrafish. The results indicated that microplastics (MPs) in combination with pesticides reduced the content of glutathione. The gene expression of cyp1a showed significant decreased in zebrafish after treatment with ABM alone and in combination with microplastic, while the expression of vtg gene was significantly increased in samples treated with combination of ABM and MPs. Analysis of antioxidant activity revealed that glutathione content, glutathione peroxidase and DPPH activity were not affected by MPs when applied alone, but it increased the total antioxidant capacity and also inhibited the superoxide dismutase (SOD) activity. ABM had no effect on glutathione peroxidase and DPPH activity, but it reduced SOD activity. Also, the combination of microplastic and toxin increased glutathione peroxidase activity and decreased the glutathione content, SOD and DPPH activity. In general, the result of this study demonstrated that the effect of MPs cannot be ignored in increasing the toxicity of other pollutants.

Changes in the Hepatic Expression of Three Vitellogenin Subtype Genes During Ovarian Development in Female White-Edged Rockfish (Sebastes taczanowskii).

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-17β 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.

Effects of cadmium on the female reproductive axis of a Neotropical fish

Metals are one of the contaminants released from the increase of anthropic activities. They can be classified as endocrine disruptors once they can affect the reproductive parameters of different organisms. The aim of the study was to evaluate the potential effects of cadmium on regulatory reproduction axis (Hypothalamic-Pituitary-Gonadal-Liver, the HPGL axis) in females of Rhamdia quelen exposed to nominal concentrations of 0.1; 1; 10 and 100 μg.L−1 of cadmium. After 15 days, tissues were collected for hormonal quantification, brain aromatase (cyp19a1b), hepatic vitellogenin (vtg) gene expression, and biomarkers analysis. Cadmium was quantified in water, gonad and liver samples. The plasma levels of estradiol, testosterone and gonad and hepatosomatic indexes did not changed after Cd exposure. The cyp19a1b was not different among the groups. Cadmium was detected at higher concentrations in the liver compared to the gonads. No genotoxicity was observed, only erythrocytes nuclear alterations. Metallothionein was reduced at 10 μg.L−1 in the liver and 10 and 100 μg.L−1 in the gonad. Hepatic superoxide dismutase activity increased and this can lead to a hydrogen peroxide increase, one of reactive oxygen species. This increase without a compensation of other enzymes of the antioxidant system can lead to lipoperoxidation, as occurred at 100 μg.L−1. Hepatic vitellogenin gene expression increased as well as the injury index at 0,1 and 100 μg.L−1. The tested cadmium concentrations have been found in the freshwater ecosystems and can affect the female reproductive regulation axis HPGL of the Neotropical species R. quelen.

The vitellogenin genes in Cynops orientalis: New insights on the evolution of the vtg gene family in amphibians.

The vitellogenins (Vtgs) are glycolipophosphoproteins that play a key role in constituting nutritional reserves for embryo development in nonmammalian vertebrates. However, additional functional roles have been evidenced. These vtg genes are present in multiple copies, different in number and sequences in various vertebrate lineages. The comprehension of the vtg gene family evolutionary history remains a matter of intense interrogation for this field of research. In tetrapods, information about vtg genes are limited to few taxa. Up to date concerning amphibians, detailed studies have been conducted only in Anura. Therefore, in this study, to further increase knowledge about vtg genes in Amphibia class, the urodele Cynops orientalis (Amphibia: Caudata) was analyzed and four complete vtg sequences were obtained. Moreover, genomic data available for the caecilians Microcaecilia unicolor and Rhinatrema bivittatum (Amphibia: Gymnophiona) were also included. In these amphibians, our findings evidenced the presence of a vtgI sequence ortholog to that of tetrapods, absent in Anura. Moreover, microsyntenic, phylogenetic, and gene conversion analyses allowed postulating two hypotheses to explain the complex evolutionary history of this gene family.

Extra-Fortification of Zinc Upsets Vitellogenin Gene Expression and Antioxidant Status in Female of Clarias magur brooders.

The present experiment was designed to evaluate the effect of graded level of zinc on Vitellogenin gene (Vtg) expression and antioxidant enzymes in threatened catfish, Clarias magur (C. magur). One hundred and eighty female C. magur with an average weight of 145 ± 5g were allocated in twelve cemented tanks with dimension 4.5 × 2 × 1m for a period of 60days. Fish were distributed in four groups with three replicates following the completely randomised design. The first group treated as control (C) fed with basal diet contained normal zinc level, and remaining groups were fed with basal diets having 50, 200 and 300mg/kg zinc acetate and treated as T1, T2 and T3 respectively. To evaluate the effect of dietary zinc supplementation on Vtg gene expression, three sampling were carried out, I sampling (April, before starting the experimental trail), II sampling (May, after 1month of feeding trail) and III sampling (June before breeding season). In the present study, a dose-dependent relationship between Vtg gene expression and zinc inclusion in the diet of threatened catfish, C. magur, was reported. Vtg gene expression increased in all groups from I sampling to II sampling but the highest Vtg gene expression was found in T1 group and the lowest in T3 group at II sampling. Vtg gene expression among the treatments differs significantly (P < 0.05) in each sampling. Accumulation of zinc was measured by Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES) in C. magur and it was reported that the significantly higher (P < 0.05) zinc was accumulated in the liver and ovary of T3 group as compared to other groups. The antioxidant enzyme activities (superoxide dismutase, SOD, catalase and GST) were also measured in different tissues (liver, gill and ovary) to evaluate the effect of extra-supplementation of zinc on the antioxidant status. In T3 group, SOD, catalase and GST activities were significantly higher than those in other groups. In the current study, serum glucose level was also measured and it was found in increasing trend with inclusion of zinc in the diet of C. magur. In the present study, it can be concluded that the zinc exhibits beneficial effect only up to 50mg/kg. Thus, it is concluded that supplementation of zinc at 200mg/kg or more disrupts Vtg gene expression and antioxidant status.