Vitellogenin Gene Expression Research Articles

Pacific White Shrimp (Litopenaeus vannamei) Vitellogenesis-Inhibiting Hormone (VIH) Is Predominantly Expressed in the Brain and Negatively Regulates Hepatopancreatic Vitellogenin (VTG) Gene Expression1

Ovarian maturation in crustaceans is temporally orchestrated by two processes: oogenesis and vitellogenesis. The peptide hormone vitellogenesis-inhibiting hormone (VIH), by far the most potent negative regulator of crustacean reproduction known, critically modulates crustacean ovarian maturation by suppressing vitellogenin (VTG) synthesis. In this study, cDNA encoding VIH was cloned from the eyestalk of Pacific white shrimp, Litopenaeus vannamei, a highly significant commercial culture species. Phylogenetic analysis suggests that L. vannamei VIH (lvVIH) can be classified as a member of the type II crustacean hyperglycemic hormone family. Northern blot and RT-PCR results reveal that both the brain and eyestalk were the major sources for lvVIH mRNA expression. In in vitro experiments on primary culture of shrimp hepatopancreatic cells, it was confirmed that some endogenous inhibitory factors existed in L. vannamei hemolymph, brain, and eyestalk that suppressed hepatopancreatic VTG gene expression. Purified recombinant lvVIH protein was effective in inhibiting VTG mRNA expression in both in vitro primary hepatopancreatic cell culture and in vivo injection experiments. Injection of recombinant VIH could also reverse ovarian growth induced by eyestalk ablation. Furthermore, unilateral eyestalk ablation reduced the mRNA level of lvVIH in the brain but not in the remaining contralateral eyestalk. Our study, as a whole, provides new insights on VIH regulation of shrimp reproduction: 1) the brain and eyestalk are both important sites of VIH expression and therefore possible coregulators of hepatopancreatic VTG mRNA expression and 2) eyestalk ablation could increase hepatopancreatic VTG expression by transcriptionally abolishing eyestalk-derived VIH and diminishing brain-derived VIH.

Tradeoff between reproduction and resistance evolution to Bt-toxin in Helicoverpa armigera: regulated by vitellogenin gene expression

Evolution of resistance to insecticides usually has fitness tradeoffs associated with adaptation to the stress. The basic regulation mechanism of tradeoff between reproduction and resistance evolution to Bacillus thuringiensis (Bt) toxin in the cotton bollworm, Helicoverpa armigera (Ha), based on the vitellogenin (Vg) gene expression was analyzed here. The full-length cDNA of the Vg gene HaVg (JX504706) was cloned and identified. HaVg has 5704 base pairs (bp) with an open reading frame (ORF) of 5265 bp, which encoded 1756 amino acid protein with a predicted molecular mass of 197.28 kDa and a proposed isoelectric point of 8.74. Sequence alignment analysis indicated that the amino acid sequence of HaVg contained all of the conserved domains detected in the Vgs of the other insects and had a high similarity with the Vgs of the Lepidoptera insects, especially Noctuidae. The resistance level to Cry1Ac Bt toxin and relative HaVg mRNA expression levels among the following four groups: Cry1Ac-susceptible strain (96S), Cry1Ac-resistant strain fed on artificial diet with Bt toxin for 135 generations (BtR stands for the Cry1Ac Bt resistance), progeny of the Cry1Ac-resistant strain with a non-Bt-toxin artificial diet for 38 generations (CK1) and the direct descendants of the 135th-generation resistant larvae which were fed on an artificial diet without the Cry1Ac protein (CK2) were analyzed. Compared with the 96S strain, the resistance ratios of the BtR strain, the CK1 strain and the CK2 strain were 2917.15-, 2.15- and 2037.67-fold, respectively. The maximum relative HaVg mRNA expression levels of the BtR strain were approximately 50% less than that of the 96S strain, and the coming of maximum expression was delayed for approximately 4 days. The overall trend of the HaVg mRNA expression levels in the CK1 strain was similar to that in the 96S strain, and the overall trend of the HaVg mRNA expression levels in the CK2 strain was similar to that in the BtR strain. Our results suggest that the changes in reproduction due to the Bt-toxin resistance evolution in the BtR strain may be regulated by the Vg gene expression. The down-regulation of HaVg at the early stages resulted in a period of delayed reproduction and decreased fecundity in the BtR strain. This performance disappeared when the Bt-toxin selection pressure was lost.

FEEDING ON A BEGOMOVIRUS‐INFECTED PLANT ENHANCES FECUNDITY VIA INCREASED EXPRESSION OF AN INSULIN‐LIKE PEPTIDE IN THE WHITEFLY, MEAM1

The Middle East-Minor 1 cryptic species (MEAM1), Bemisia tabaci (Gennadius) is a globally invasive pest. It spreads widely due to its high fecundity and mutualistic interactions with the virus they vector. Feeding on virus (tomato yellow leaf curl China virus, TYLCCNV)-infected host plants improves their fecundity, however, the key factor regulating the signaling transduction in reproduction of whitefly remains to be identified. Here, we cloned a full length cDNA encoding an insulin-like peptide in MEAM1 (BtILP1) and investigated its expression profile, functions, and the expression induced by feeding on virus-infected tobacco plants. The full length cDNA of BtILP1 was 590 bps and encoded an open reading frame containing 149 amino acid residues. Multiple sequences alignment results showed BtILP1 contained the structural features typical of the insulin family. Expression dynamics associated with development showed the expression level of BtILP1 peaked at 5 days posteclosion (PE). During 1 to 3 days PE, BtILP1 was expressed highly in the head and abdomen of female adults and highly in the head during 5 to 7 days PE. Knockdown of the BtILP1 expression also impaired vitellogenin gene expression at both transcript and protein levels. Downregulating BtILP1 expression decreased fecundity of female adults and hatching rate of eggs. Feeding on virus-infected tobacco increased BtILP1 expression in MEAM1 female adults. We infer feeding on begomovirus-infected tobacco enhances the reproduction of MEAM1 by inducing BtILP1 expression. Our results give a new sight into the mutualistic interactions between virus and its insect vector.

Effects of 17α-ethinyl estradiol exposure on estrogen receptors α and β and vitellogenins A, B and C mRNA expression in the liver of sand goby (Pomatoschistus minutus)

This study aims to characterize the estrogen receptor (er) in sand goby (Pomatoschistus minutus) and determine the temporal effects of 17α-ethinyl estradiol (EE2) on erα and vitellogenin (vtg) gene expression in males. Two partial cDNA sequences (erα and erβ1) are presented showing conserved structural features with ers of other species. Transcript levels for both ers were low in control fish but EE2 exposure (11 ng/L, for 29 days) increased both to a pattern similar to vitellogenic females. The relative expression of three vtg genes (vtga, vtgb and vtgc) along with erα was determined in control and male fish exposed to EE2 (11 ng/L) at multiple time-points over 29 days. All four transcripts were significantly induced due to exposure and expression rose during the time course with distinct temporal patterns and vtga reached a substantially higher level at the end of the time course coinciding with rapid elevation in erα expression.

Evaluation of Cage Designs and Feeding Regimes for Honey Bee (Hymenoptera: Apidae) Laboratory Experiments

The aim of this study was to improve cage systems for maintaining adult honey bee (Apis mellifera L.) workers under in vitro laboratory conditions. To achieve this goal, we experimentally evaluated the impact of different cages, developed by scientists of the international research network COLOSS (Prevention of honey bee COlony LOSSes), on the physiology and survival of honey bees. We identified three cages that promoted good survival of honey bees. The bees from cages that exhibited greater survival had relatively lower titers of deformed wing virus, suggesting that deformed wing virus is a significant marker reflecting stress level and health status of the host. We also determined that a leak- and drip-proof feeder was an integral part of a cage system and a feeder modified from a 20-ml plastic syringe displayed the best result in providing steady food supply to bees. Finally, we also demonstrated that the addition of protein to the bees' diet could significantly increase the level ofvitellogenin gene expression and improve bees' survival. This international collaborative study represents a critical step toward improvement of cage designs and feeding regimes for honey bee laboratory experiments.

Juvenile hormone signaling during reproduction and development of the linden bug, Pyrrhocoris apterus

Juvenile hormone (JH), a sesquiterpenoid produced by the insect corpus allatum gland (CA), prevents metamorphosis in larvae and stimulates vitellogenesis in adult females. Whether the same JH signaling pathway regulates both processes is presently unknown. Here, we employ the robust JH response during reproduction and development of the linden bug, Pyrrhocoris apterus, to compare the function of key JH-signaling genes encoding the JH receptor, Methoprene-tolerant (Met), its binding partner Taiman (Tai), and a JH-inducible protein, Krüppel-homolog 1 (Kr-h1). RNA interference (RNAi) with Met or Tai, but not Kr-h1, blocked ovarian development and suppressed vitellogenin gene expression in the fat body of females raised under reproduction-inducing conditions. Loss of Met and Tai matched the effects of CA ablation or the natural absence of JH during reproductive diapause. Stimulation of vitellogenesis by treatment of diapausing females with a JH mimic methoprene also required both Met and Tai in the fat body, whereas Kr-h1 RNAi had no effect. Therefore, the Met-Tai complex likely functions as a JH receptor during vitellogenesis. In contrast to Met and Kr-h1 that are both required for JH to prevent precocious metamorphosis in P. apterus larvae, removal of Tai disrupted larval ecdysis without causing premature adult development. Our results show that while Met operates during metamorphosis in larvae and reproduction in adult females, its partner Tai is only required for the latter. The diverse functions of JH thus likely rely on a common receptor whose actions are modulated by distinct components.

A real-time PCR assay for differential expression of vitellogenin I and II genes in the liver of the sentinel fish species Lipophrys pholis

The recent advances in molecular biology techniques have prompted the use of vitellogenin (VTG) gene expression as a sensitive and reliable indicator of estrogenic chemicals (EC) exposure. However, data on the dynamic response of the different VTGs genes upon EC exposure is still poorly understood, particularly in sentinel fish species used in field monitoring studies. Hence, the present study aimed at developing a sensitive real-time PCR assay for determining the response of VTG I and II in the recently proposed marine sentinel species Lipophrys pholis upon exposure to the model EC 17α-ethinylestradiol (EE2). The findings of the laboratory study indicate that L. pholis VTG I proved to be not only more inducible but also more sensitive to EE2 exposure than VTG II, for the same range of concentrations. In fact, VTG I gene induction was 475-fold higher than VTG II at 15 ng/L EE2, and 13-fold at 5 ng/L EE2. Overall, the findings of the present study indicate that in the field, expression of VTG I in L. pholis should be preferentially used in the screening of EC exposure because of its higher sensitivity. Furthermore, the present study favors L. pholis integration in monitoring programs associated with EC’s pollution within the European water policy legislation.

Effects of estrone on the early life stages and expression of vitellogenin and estrogen receptor genes of Japanese medaka (Oryzias latipes)

The fertilized eggs of Japanese medaka (Oryzias latipes) were exposed to estrone (E1) at 5–5000ngL−1 for 15d, and the hatched fry were exposed continuously to the same concentrations for the additional 15d. Adverse effects on hatchability, time to hatching, and gross abnormalities occurred at 50ngL−1 or above. Then the fry were divided into a continual exposure group, and a water recovery group. When the fry were exposed to E1 for another 60d, there was a decrease in the hepatosomatic index (HSI) of males and the influence disappeared in the water recovery group. The gonadosonatic index (GSI) of females at 500ngL−1 decreased significantly in another 60d exposure. While the fry were maintained in dechlorinated tap water for 60d, a significant decrease in female GSI was observed at 50ngL−1 or above. An increased GSI was found in males in both continual exposure and water recovery groups at all E1 treatments. Quantitative RT-PCR showed that vitellogenin-I (Vtg-I) gene expressions in the female liver were significantly down-regulated at 50ngL−1 in the continual exposure group, and at 500ngL−1 in the water recovery group, while male Vtg-I genes were significantly up-regulated for all E1 treatments. In addition, all E1 treatments caused sex reversal of males. These results suggest that E1 at 5ngL−1 or above have unrecoverable impacts on the gonadal growth and development of medaka, even if only early life stages were exposed to E1.

Reproductive toxicity effects of 4-nonylphenol with known endocrine disrupting effects and induction of vitellogenin gene expression in silkworm, Bombyx mori

4-Nonylphenol (4-NP) a known endocrine disrupting chemical is a persistent environmental contaminant. However, the mechanism of reproductive toxicity caused by 4-NP is still largely unresolved in invertebrates. In this study, Bombyx mori larvae were constantly fed 4-NP at concentrations ranging from 0.05 to 0.4gkg−1, reproductive toxicity and induction of vitellogenin gene (Vg) expression were investigated in this organism which is an ideal lepidopteran model insect. The results showed that gonad development was retarded and maturity was decreased in both male and female pupae, while the sex ratio was unaffected by 4-NP exposure. In the 4-NP exposed animals, the corresponding egg yolk protein, vitellin, involved in energy reserves for embryonic development in oviparous animals, was present in the testis of male pupae, and the mRNA transcript of the Vg gene was detected in the fat body, a specific organ of Vg synthesis, which is normally silent in males. In addition, expression of the Vg gene was up-regulated in the fat body of female pupae and adults, while the protein was decreased in developing eggs. Furthermore, expression of the ecdysone receptor gene (EcR) in the ovaries of pupae was down-regulated, suggested that the transport of Vg from the fat body to developing oocytes was disturbed by 4-NP due to interference in the expression of EcR related to ecdysone activity.

Estrogen-like Effects in Male Goldfish Co-exposed to Fluoxetine and 17 Alpha-Ethinylestradiol

The antidepressant fluoxetine (FLX) and the synthetic estrogen, 17 alpha-ethinylestradiol (EE2), are present in municipal sewage discharges. To better understand possible interactions between them, male goldfish were exposed to an ethanol control or to nominal concentrations of FLX (0.54 μg/L) and EE2 (5 ng/L) alone and in combination for 14 days. Real-time reverse-transcription polymerase chain reaction was used to assess effects on hepatic gene expression and liquid chromatography tandem mass spectrometry to analyze the plasma proteome. The results showed an increase in estrogen receptor alpha (esr1) and vitellogenin (vtg) gene expression by 1.9-2.4-fold in the FLX and EE2 groups, but this did not reach statistical significance. In contrast, co-exposure up regulated esr1 and vtg gene expression by 5.5- and 5.3-fold, respectively. Fluoxetine and EE2 alone did not affect estrogen receptor beta (esr2), but the co-exposure down regulated esr2 expression by 50%. There was a significant increase in the number of plasma proteins that were related to endocrine system disorders in the FLX and FLX plus EE2 groups. The level of VTG protein was increased in the plasma from goldfish exposed to EE2, FLX, and FLX plus EE2. Our study demonstrates that low concentrations of FLX and EE2 in a simple mixture produce strong estrogen-like effects in the male goldfish.

Induction of gene responses in St. Lawrence River northern pike (Esox lucius) environmentally exposed to perfluorinated compounds

Municipal waste water effluents (MWWEs) are important sources of chemical contamination for aquatic environments. This study investigated the presence and effects of perfluorinated compounds (PFCs) in environmentally exposed northern pike (Esox lucius) collected upstream and downstream a major municipal waste water treatment plant (WWTP) in the St. Lawrence River, Canada. Twelve PFCs, including the newly detected perfluoroethylcyclohexane sulfonate (PFECHS), were quantified in fish muscle, liver, and plasma. Additionally, the expression of eight genes and the activity of three biomarkers were analyzed in fish tissues at both sites. Mean ∑PFC concentration in fish plasma collected upstream the WWTP was 185ng/g w.w. compared to 545ng/g w.w. downstream the point of release. PFECHS was quantified for the first time in St. Lawrence River fish (mean plasma concentration in MWWE fish: 5.07±4.72ng/g w.w.). Results of transcriptomic responses were tissue-specific and indicated significant up-regulation for metallothionein (MT) in blood and MT, glutathion-S-transferase (GST), superoxide dismutase (SOD), and cytochromes P450 1A1 (CYP1A1) in gill tissue of fish collected in the MWWE suggesting greater stress responses for organisms at this location. Significant relationships were found between vitellogenin (Vtg) gene expression in liver, Vtg activity in plasma and perfluorotridecanoic acid (PFTrA), perfluorotetradecanoic acid (PFTeA), and perfluorodecane sulfonate (PFDS) plasma concentrations. The possible endocrine effects of these PFCs should be further investigated.

Molecular cloning of a cDNA encoding vitellogenesis-inhibiting hormone in the whiteleg shrimp Litopenaeus vannamei and preparation of its recombinant peptide using an E. coli expression system

The cDNA encoding a precursor of Liv-SGP-G, the most abundant crustacean hyperglycemic hormone-family peptide in the sinus gland of the whiteleg shrimp Litopenaeus vannamei, was cloned by RT-PCR coupled with 5′- and 3′-RACE. The determined cDNA sequence consisted of 621 nucleotides comprising a 69-bp 5′-untranslated region, a 357-bp open reading frame, and a 195-bp 3′-untranslated region. The deduced sequence of 72 amino acid residues corresponding to mature Liv-SGP-G was completely identical to that of natural Liv-SGP-G, determined in our previous study. The recombinant Liv-SGP-G was thereafter heterologously expressed in Escherichia coli, and its vitellogenesis-inhibiting activity in ex vivo cultured ovarian fragments was examined. The recombinant peptides inhibited vitellogenin gene expression with almost the same efficacy as that of natural Liv-SGP-G purified from sinus glands.

Interaction of 17β-estradiol and ketoconazole on endocrine function in goldfish (Carassius auratus)

An understanding of the effects of toxic mixtures of endocrine disrupting chemicals (EDCs) on aquatic organisms is challenging as these organisms are exposed to multiple classes of contaminants in their natural habitat. The aim of the present study was to evaluate the interactions of two classes of EDCs, 17β-estradiol (E2) and ketoconazole (KTC), on endocrine function in male goldfish (Carassius auratus), including vitellogenesis, metabolic capability and serum steroid synthesis. Changes in vitellogenin (VTG) concentration, liver 7-ethoxyresorufin-O-deethylase (EROD) activity and circulating serum E2 level were examined. The expression of related genes was also determined using quantitative real-time polymerase chain reaction. Exposure to E2 caused a significant increase in VTG concentrations which corresponded with the gene expression of VTG and estrogen receptor (ER) in males, which were further elevated after combined exposure to E2 and KTC, indicative of a synergetic relationship. Exposure to E2 also resulted in a distinct increase in serum steroid biosynthesis and associated cytochrome P450 (CYP) aromatase expression after 10 days. However, these changes were inhibited by the presence of KTC, which acted as a steroidogenic inhibitor in fish. Moreover, KTC significantly decreased liver EROD activity and increased the related gene expression of CYP1A. However, these KTC-mediated metabolic reactions in goldfish were up-regulated following exposure to KTC in combination with E2. These findings reveal complex interactions on endocrine functions in male goldfish when exposed to multiple contaminations and may provide a better understanding of the effects of toxic mixtures.

Effects of illegal cyanide fishing on vitellogenin in the freshwater African catfish, Clarias gariepinus (Burchell, 1822)

The effects of cyanide, used in illegal fishing, on one of the most economically important Nile fishes, the African catfish (Clarias gariepinus), were studied. Cyanide impacts were evaluated in terms of biochemical, molecular and histopathological characteristics. After exposure to sublethal concentration (0.05mg/l) of potassium cyanide (KCN) for two and four weeks, GOT (glutamate oxaloacetate transaminase) was significantly increased in both male and female, while GPT (glutamate pyruvate transaminase), total plasma protein, phosphoprotein phosphorus (Vgt) in serum, vitellogenin gene expression (Vtg mRNA) and estrogen receptors (ER mRNA) were significantly decreased in female. On the other hand, male C. gariepinus showed a significant increase in Vtg and Vtg mRNA. Liver, testis and ovaries showed distinct histopathological changes. It was concluded that, cyanide caused damaging effects to fish and can cause serious disturbance in the natural reproduction and a drastic decline in fish population. Therefore, it is recommended that, the use of cyanide compounds must be prohibited to conserve the fisheries resources.

Multiple ferritins are vital to successful blood feeding and reproduction of the hard tick Haemaphysalis longicornis

Ticks are obligate hematophagous parasites and important vectors of diseases. The large amount of blood they consume contains great quantities of iron, an essential but also toxic element. The function of ferritin, an iron storage protein, and iron metabolism in ticks need to be further elucidated. Here, we investigated the function a newly identified secreted ferritin from the hard tick Haemaphysalis longicornis (HlFER2), together with the previously identified intracellular ferritin (HlFER1). Recombinant ferritins, expressed in Escherichia coli, were used for anti-serum preparation and were also assayed for iron-binding activity. RT-PCR and western blot analyses of different organs and developmental stages of the tick during blood feeding were performed. The localization of ferritins in different organs was demonstrated through an indirect immunofluorescent antibody test. RNA interference (RNAi) was performed to evaluate the importance of ferritin in blood feeding and reproduction of ticks. The midgut was also examined after RNAi using light and transmission electron microscopy. RT-PCR showed differences in gene expression in some organs and developmental stages. Interestingly, only HlFER2 was detected in the ovary during oviposition and in the egg despite the low mRNA transcript. RNAi induced a reduction in post-blood meal body weight, high mortality and decreased fecundity. The expression of vitellogenin genes was affected by silencing of ferritin. Abnormalities in digestive cells, including disrupted microvilli, and alteration of digestive activity were also observed. Taken altogether, our results show that the iron storage and protective functions of ferritin are crucial to successful blood feeding and reproduction of H. longicornis.

Effect of TBT and PAHs on CYP1A, AhR and Vitellogenin Gene Expression in the Japanese Eel, Anguilla japonica.

Gene expressions of cytochrome P4501A (CYP1A), aryl hydrocarbon receptor (AhR) and vitellogenin (Vg) by endocrine disruptors, benzo[α]pyrene (B[a]P) and tributyltin (TBT) were examined in cultured eel hepatocytes which were isolated from eels treated previously with B[a]P (10 mg/kg) or estradiol-17β (20 mg/kg) in vivo, and the relationship between CYP1A, AhR and Vg genes were studied. When the cultured eel hepatocytes were treated with B[a]P (10-6-10-5 M) the gene expressions of CYP1A and AhR were enhanced in a concentration-dependent manner. However, when treated with TBT (10-9-10-5 M) the gene expressions of CYP1A and AhR were suppressed at high concentrations (10-6-10-5 M), while having no effects at low concentrations (10-9-10-7 M). Gene expression of Vg was also suppressed by TBT in a concentration-dependent manner in cultured eel hepatocytes which was previously treated in vivo with estradiol-17β.

The ex vivo effects of eyestalk peptides on ovarian vitellogenin gene expression in the kuruma prawn Marsupenaeus japonicus

Seven major peptides belonging to the crustacean hyperglycemic hormone family were purified from the sinus gland located in the eyestalk of the kuruma prawn Marsupenaeus japonicus, and their effects on vitellogenin gene expression were examined using the ex vivo ovary incubation system. Six molecular species of crustacean hyperglycemic hormone, Pej-SGP-I, -II, -III, -V, VI, and VII, displayed significant inhibitory effects on vg expression with almost the same efficacies, whereas Pej-SGP-IV (known as molt-inhibiting hormone) did not. Two chromatophorotropic peptides, red pigment-concentrating hormone and pigment-dispersing hormone, which were also present in the sinus glands, did not have a clear effect on the gene expression levels in this incubation system. These results suggest that the six crustacean hyperglycemic hormones are potentially capable of acting as vitellogenesis-inhibiting hormones in M. japonicus.

Suppression of fecundity, Nlvg gene expression and vitellin content in Nilaparvata lugens Stål (Hemiptera: Delphacidae) adult females exposed to indoxacarb and chlorantraniliprole

The brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), is a classic example of a pest that experiences insecticide-induced fecundity. The insecticides indoxacarb and chlorantraniliprole were recently developed with high insecticidal activity in the control of rice yellow stem borer, Tryporyza incertulas (walker) and rice leaffolder, Cnaphalocrocis medinalis (Guenee). We examined the changes in the fecundity of N. lugens, the vitellogenin (Nlvg) gene expression and the vitellin contents of adult females exposed to these new insecticides. Our findings showed that indoxacarb and chlorantraniliprole significantly suppressed the fecundity of the females and the expression of Nlvg mRNA. Treatment with 10, 20, 40 and 80mg/L indoxacarb decreased Nlvg mRNA expression by 35.4%, 74.4%, 79.6% and 61.9%, respectively, and treatment with 7.5, 15, 30, 60 and 80mg/L chlorantraniliprole decreased Nlvg mRNA expression by 79.3%, 40.2%, 40.0%, 79.8% and 51.8%, respectively. The vitellin contents in both the ovaries and fat bodies of the adult females also decreased, indicating that the two insecticides suppressed the synthesis of vitellin in the fat bodies and the transfer of vitellin to the ovaries. This decrease in the vitellin contents of the ovaries may be attributed to the low Nlvg expression, as there was a positive correlation between the vitellin content and the expression of this gene, indicating that these insecticides influenced the fecundity of N. lugens by affecting the expression of Nlvg mRNA. The study of the effects of indoxacarb and chlorantraniliprole on fecundity should have significance for the wide usage in rice plants.

Effects of butachlor on estrogen receptor, vitellogenin and P450 aromatase gene expression in the early life stage of zebrafish

Butachlor has adverse effects on fecundity and disrupts sex hormone homeostasis in adult zebrafish, but the underlying molecular mechanisms are still unclear. In the present study, zebrafish (Danio rerio) embryos were exposed to various concentrations of butachlor from 2 h post-fertilization (hpf) to 30 days post-fertilization (dpf). The transcription of genes involved estrogen receptors (ERα, ERβ1 and ERβ2), vitellogenins (VTG I and II), and cytochrome P450 aromatase (CYP19a) was analyzed by real-time quantitative PCR. The results showed that there was no significant alteration in the expression of VTGI, ERα, ERβ1, ERβ2 and CYP19a after 30 days of butachlor exposure, whereas the transcription of VTG II gene was significantly up-regulated in zebrafish exposed to 100 μg/L butachlor. It is suggested that butachlor may be a weak estrogen, and more endpoints need to be investigated to assess the effects of butachlor on the hypothalamus–pituitary–gonadal axis of zebrafish.

Cloning and expression of vitellogenin (Vg) gene and its correlations with total carotenoids content and total antioxidant capacity in noble scallop Chlamys nobilis (Bivalve: Pectinidae)

As a nonpolar molecular carrier and a storage protein, vitellogenin (Vg) cannot only combine and transfer lipids, proteins, vitamin and carotenoids to oocytes during the oogenesis, but also be linked with the host immune defense. In this study, the full-length cDNA encoding Vg in noble scallop Chlamys nobilis was cloned. The complete Vg cDNA consists of 7760 nucleotides with an open reading frame encoding 2289 amino acid residues. Phylogenetic analysis of Vg gene revealed that Ch. nobilis was clustered together firstly with its sister species Chlamys farreri and another scallop Mizuhopecten yessoensis, then with other mollusks such as oyster and abalone, and finally with vertebrates. Tissue-specific checking results indicated that Vg gene was only expressed in the ovary. At different gonadal development stages, both orange and brown shell color scallops show the same trend that the amount of Vg mRNA expression kept at a high level at the growing stage, then dramatically decreased at the mature stage, and finally resumed to higher level at the post-spawning stage. Another very important finding is that significantly positive correlations existed between the Vg gene expression level and total carotenoids content, as well as total antioxidant capacity. The relationship among them needs further study.