Female Scallops Research Articles

Annual variation in thermal and structural properties of yesso scallops (Mizuhopecten yessoensis) adductor muscle

The annual changes of the scallop’s adductor muscle in Ca2+-ATPase activity, chymotryptic digestion, endogenous fluorescence spectra, sulfhydryl content, and surface hydrophobicity were studied. Ca2+-ATPase activity peaked in February and reached its minimum in August for males and in July for females. However, no significant differences were observed between males and females throughout the year. The inactivation rate of Ca2+-ATPase activity increased by 17- to 36-fold when the temperature rose from 38°C to 45°C. The ratio of free myosin to bound myosin was approximately 5:4 at 0.5 M KCl and changed to 5:1 at 1.0 M, as determined by modeling. Chymotryptic digestion demonstrated that scallop myosin could be cleaved into S-1/rod portions at 0.1 M and HMM/LMM at 0.5 M KCl. Furthermore, no significant seasonal variations were observed in chymotryptic digestion patterns, endogenous fluorescence, surface hydrophobicity, or sulfhydryl content. In conclusion, the structure and thermal stability of both male and female scallops remained stable throughout the year, making them suitable for processing and preservation.

Effects of Benzo[a]pyrene on Food Metabolism and Reproductive Endocrine and Ovarian Development in Female Scallop Chlamys farreri at Different Reproductive Stages.

Benzo[a]pyrene (B[a]P), a polycyclic aromatic hydrocarbon (PAH) with the most carcinogenic effects of all the PAHs, has multiple toxic effects on marine bivalves. We investigated the interference mechanism of B[a]P on food metabolism (sugars, proteins, and sugars), and on reproductive endocrine and ovarian development in female scallops (Chlamys farreri). Scallops were exposed to different concentrations of B[a]P concentrations of 0, 0.38, 3.8, and 38 μg/L throughout gonadal development. Total cholesterol and triglyceride contents in the digestive glands were increased, and their synthesis genes were upregulated. The plasma glucose contents decreased with the inhibition of glycogen synthesis genes and the induction of glycolysis genes in the digestive gland. The results showed that B[a]P had endocrine-disrupting effects on scallops, that it negatively affected genes related to ovarian cell proliferation, sex differentiation, and egg development, and that it caused damage to ovarian tissue. Our findings supplement the information on B[a]P disruption in gonadal development of marine bivalves. Environ Toxicol Chem 2024;43:748-761. © 2023 SETAC.

Insights into mechanism of DNA damage and repair-apoptosis in digestive gland of female scallop Chlamys farreri under benzo[a]pyrene exposure during reproductive stage

As one of the most carcinogenic persistent organic pollutants (POPs), benzo[a]pyrene (B [a]P) brings high toxicity to marine bivalves. Digestive gland is the most important metabolism-related organ of aquatic animals. This study conducted the digestive gland transcriptome of Chlamys farreri under B[a]P treatment at reproductive stages. And the reproductive-stage dependence metabolism-DNA repair-apoptosis process of scallops under 0, 0.04, 0.4 and 4 μg/L B[a]P was studied by qRT-PCR. The results demonstrated that the detoxification metabolism was disturbed after ovulation except for CYP3A4. In antioxidant system, antioxidant enzyme CAT and GPX, and GGT1 (one of the non-enzymatic antioxidants synthesis gene) continuously served the function of antioxidant defense. Three types of DNA repair were activated under B[a]P stress, however, DNA strand breaks were still serious. B[a]P exposure weakened death receptor pathway as well as enhanced mitochondrial pathway, surprisingly suppressing apoptosis in scallops. In addition, ten indicators were screened by Spearman correlation analysis. This study will provide sound theoretical basis for bivalve toxicology and contribute to the biomonitoring of marine POPs pollution.

Identification and characterization of gonadotropin-releasing hormone (GnRH) in Zhikong scallop Chlamys farreri during gonadal development.

Gonadotropin-releasing hormone (GnRH) controls synthesis of sex steroid hormones through hypothalamic-pituitary-gonadal (HPG) axis in vertebrates. But in mollusks, research on neuroendocrine control of gonadal function, such as the function of GnRH during gonadal development is limited. In this study, we investigated the morphology and structure of the nerve ganglia of Zhikong scallop Chlamys farreri by physiological and histological observations. We also cloned the ORF and studied the expression patterns of GnRH in the scallop. Tissue expression analysis showed that GnRH was highly expressed in parietovisceral ganglion (PVG). The in situ hybridization result further confirmed that GnRH mRNA only distributed in some good-sized neurons in the posterior lobe (PL) and some pint-sized neurons in the lateral lobe (LL). In addition, by examining the expression of GnRH during gonadal development in ganglia, we found GnRH displayed higher expression in the female scallops, and showed significant high expression at the growing stage of female scallops in PVG. This study would contribute to gaining insight into the mechanism underlying reproduction regulation by GnRH in the scallop and help to provide a better understanding of reproductive neuroendocrine in mollusks.

Reproductive toxicity induced by benzo[a]pyrene exposure: first exploration highlighting the multi-stage molecular mechanism in female scallop Chlamys farreri.

Reproductive toxicity induced by benzo[a]pyrene (B[a]P) exposure has received great ecotoxicological concerns. However, huge gaps on the molecular mechanism still exist in bivalves. In this study, reproduction-related indicators were investigated in female scallops Chlamys farreri during life cycle of proliferative, growth, mature, and spawn stages, under gradient concentrations of B[a]P at 0, 0.04, 0.4, and 4μg/L. Meanwhile, a multi-stage ovarian transcriptome analysis under 4μg/L B[a]P exposure was also conducted to elucidate the potential molecular mechanisms. The results indicated that life-cycle exposure to 0.4 and 4μg/L B[a]P significantly decreased GSI and sex steroid levels. Even 0.04μg/L B[a]P could play the adverse role in DNA integrity at the mature and spawn stages. Ovarian histological sections showed that B[a]P inhibited the maturation and release of oocytes. Through the functional enrichment analysis of differentially expressed genes (DEGs) from transcriptome data, 18 genes involved in endocrine disruption effects, DNA damage and repair, and oogenesis were selected and further determined by qRT-PCR. The downregulation of genes involved in steroidogenic and estrogen signaling pathways indicated that B[a]P could cause endocrine disruption through both receptor-dependent and receptor-independent pathways. The variations of gene expressions involved in DNA single-strand break and repair implied the presence of toxic mechanisms similar with vertebrates. Additionally, the changes of gene expressions of cell cycle, apoptosis, and cell adhesion suggested that exposure to B[a]P possibly caused the reproductive toxicity effects by affecting oogenesis. Taken together, this study was a pioneer in combining genome-wide transcriptomic analysis with its corresponding reproductive indicators (GSI, sex steroid levels, DNA single-strand break, and histological sections) to explore the bivalves' toxic mechanisms under B[a]P exposure. Meanwhile, some genes involved in estrogen signaling pathway and DNA damage were firstly analyzed in bivalves, and the expression data might be useful in establishing new hypotheses and discovering new biomarkers for marine biomonitoring.

Insights into disruption of lipid metabolism in digestive gland of female scallop Chlamys farreri under B[a]P exposure

Lipids are the main energy support during gametogenesis. Digestive gland is the key organ of aquatic animal metabolism for storing nutrition and supplying energy. It participates in a variety of life activities (such as growth, digestion, immunity, and reproduction). Nutrients stored in digestive glands, especially lipids, provide energy for reproductive behaviors such as gametogenesis and ovulation. A large number of studies have confirmed the accumulation of lipids from digestive gland to gonad during gametogenesis. At present, the research on the interference mechanism of persistent organic pollutants (POPs) on lipid metabolism of aquatic animals and the adaptive response of aquatic animals to POPs stress focus on biochemical levels or a few genes. The potential molecular mechanism of lipid metabolism interference needs to be further studied. In addition, as an important stage of aquatic animals, the reproductive period is a vigorous period of lipid metabolism. However, at present, there is no report on the molecular mechanism of POPs interfering with the lipid metabolism of the digestive gland in the reproductive process of aquatic animals. In this study, female scallop C. farreri was cultured in natural seawater and exposed to 4 μg/L B[a]P in seawater. Transcriptome analysis of digestive glands at multiple stages (proliferative stage, growth stage, mature stage and spawn stage) was performed, and iPath pathway analysis was used to analyze lipid metabolism pathways and differential genes. The interference mechanism of lipid metabolism in bivalves during reproductive period was revealed. This study will provide valuable genomic information on the role of digestive glands in lipid metabolism and reproduction of C. farreri, and will contribute to further functional genomics of bivalves and other closely related species.

Cell culture and genetic transfection methods for the Japanese scallop, Patinopecten yessoensis.

Cell cultures can simplify assays of biological phenomena; therefore, cell culture systems have been established for many species, even invertebrates. However, there are few primary culture systems from marine invertebrates that can be maintained long term. The Japanese scallop, Patinopecten yessoensis, is a marine bivalve. Cell culture systems for the scallop have only been established for a few organ‐derived cell types and for embryonic cells. We developed a primary culture system for cells from male and female scallop gonads, hepatopancreas, and adductor muscle by utilizing culture conditions closer to those in nature, with regard to temperature, osmolarity, and nutrition. Primary cultured female gonadal cells were maintained for more than 1 month and had potential for proliferation. Furthermore, a genetic transfection system was attempted using a scallop‐derived promoter and a lipofection reagent. GFP‐positive cells were detected in the attempt. These technical developments would promote our understanding of biochemical mechanisms in scallops as well as providing clues for establishment of immortalized molluscan cell lines.

Carotenoids content and lipid nutritional quality variation in tissues of male and female polymorphic (golden and brown) noble scallops Chlamys nobilis

The noble scallop Chlamys nobilis is one of the most important cultured shellfish in Southern China, known for its rich total carotenoids (TCC) and polyunsaturated fatty acids (PUFA). However, carotenoid and lipid nutritional quality of C. nobilis are greatly affected by many factors, including season, sex, and color of C. nobilis. Therefore, this study aims to assess the TCC and lipid nutritional quality of tissues of male and female, golden and brown scallops. The results of the this study revealed that golden scallops generally have much higher TCC, total lipid content (TLC) and PUFA than brown scallops, while female scallops generally have higher TCC, but lower TLC and PUFA than male scallops. In conclusion, golden scallops are more nutritious than brown scallops. Female golden scallops may be more attractive to consumers interested in high natural antioxidant (TCC), while male golden scallops may be more popular for consumers interested in high quality lipids (TLC and PUFA). These data will help consumers determine the best dietary choices based on their nutritional interests.

Temporal transcriptome analysis in female scallop Chlamys farreri: First molecular insights into the disturbing mechanism on lipid metabolism of reproductive-stage dependence under benzo[a]pyrene exposure

Polycyclic aromatic hydrocarbons (PAHs) are one of the most widespread persistent organic pollutants (POPs) in marine environment. Benzo[a]pyrene (B[a]P), the most toxic carcinogen of PAHs, is widely studied as a representative that interferes with lipid metabolism. However, the underlying molecular mechanisms of lipid metabolism by B[a]P interference towards bivalve, one of the marine-pollution bio-indicators have not been elucidated yet, especially during gonadal development which is closely associated with lipids. In this study, female scallops Chlamys farreri were cultured with natural and 4 μg/L B[a]P exposed seawater, respectively, and a multi-stage (proliferative, growth, mature, and spawn stage) ovarian transcriptome profiling was performed to decipher the reproductive stage-dependence disturbing mechanisms on lipid metabolism caused by B[a]P in bivalves. The results revealed the potential molecular mechanism of B[a]P-induced triglycerides (TGs) accumulation, which probably resulted from the collaboration of promoting synthesis and inhibiting metabolization of TGs, notably, this mechanism also occurred at spawn stage. Correspondingly, B[a]P and TGs contents measured in ovary offered direct biochemical evidences for the interference effects and stage-dependent accumulation patterns of B[a]P. Moreover, the gene expressions of fatty acids synthesis related enzymes were down-regulated cooperatively, illustrating the molecular compensatory mechanism that reduced susceptibility from oxidative damage. And these results further emphasized the important role of prostaglandins (PGs) in immune response mediated by arachidonic acid metabolism. In addition, this study explored the underlying molecular mechanism affected by B[a]P on sterol metabolism, which possibly posed a threat to normal reproductive functions in bivalves. Taken together, our findings filled the gap of the stage-dependent interference molecular mechanisms on lipid metabolism behind bivalves, and provided a new perspective for investigating the adaptive mechanisms of bivalves under POPs stress.

Characterizing transcriptome in female scallop Chlamys farreri provides new insights into the molecular mechanisms of reproductive regulation during ovarian development and spawn

Bivalve mollusks are descendants of an early-Cambrian lineage and have successfully evolved unique strategies for reproduction. Nonetheless, the molecular mechanisms underlying reproductive regulation in mollusks remain to be elucidated. In this study, transcriptomes of ovary at four reproductive stages in female Chlamys farreri were characterized by RNA-Seq. Regarding signaling pathways, ECM-receptor interaction pathway, mTOR signaling pathway, Fanconi anemia pathway, FoxO signaling pathway, Wnt signaling pathway and Hedgehog signaling pathway were enriched during ovarian development processes. In addition, pathways related to energy metabolism such as Nitrogen metabolism and Arachidonic acid metabolism were enriched at spawn stage. Interestingly, Neuroactive ligand-receptor interaction was significantly enriched involved in ovarian development and spawn, and indicated the potential functions of nervous system on reproductive regulation in C. farreri. What's more, this study identified and characterized fourteen genes involved in "sex hormones synthesis and regulation", "ovarian development and spawn" and "maternal immunity" during the four reproductive stages in C. farreri. We determined that CYP17 uniquely affected gamete release by influencing the physiological balance among the steroid hormones and showed that receptors of the 5-HT and GABA neurotransmitters were tightly associated with ovarian maturation. Furthermore, to the best of our knowledge, this is the first study to report the maternal effect gene Zar1 in bivalve mollusks, likewise the maternal immunity genes displayed coordinated and cooperative expression during reproductive periods, which strengthened the environmental adaptation mechanisms of bivalves. Taken together, this study provides the first dynamic transcriptomic analysis of C. farreri at four key reproductive stages, which will assist in revealing the molecular mechanisms underlying bivalves on reproductive regulation in ovarian development and spawn.

Benzo[a]pyrene exposure induced reproductive endocrine-disrupting effects via the steroidogenic pathway and estrogen signaling pathway in female scallop Chlamys farreri

Benzo[a]pyrene (B[a]P), as one of the typical polycyclic aromatic hydrocarbons and environmental contaminants, may cause endocrine disrupting effects and reproductive impairments in bivalves. However, the molecular mechanisms are still not fully understood. In this study, three reproductive stages (proliferative stage, growing stage and mature stage) of female scallops Chlamys farreri were exposed to B[a]P at 0, 0.38 and 3.8 μg/L. The present study determined the adverse effects of B[a]P on gonadosomatic index, circulating hormone concentrations, endocrine-associated gene expression and ovarian histology. Significant decrease in sex hormones including progesterone (P), testosterone (T) and 17β-estradiol (E2), was observed in B[a]P-treated C. farreri at growing stage and mature stage. These effects were associated with down-regulated expression of steroidogenic enzymes, including 3β-HSD, CYP17 and 17β-HSD, which were regulated by the upstream adenylate cyclase (Adcy) - protein kinase (PKA) signaling pathway. Ovarian transcript levels of estrogen receptor (ER) and caveolin-1 (cav-1) were decreased in B[a]P-treated C. farreri. Vitellogenin (Vtg), an estrogen-mediated gene involved in ovarian development, was down-regulated by B[a]P. Furthermore, ovarian histology was investigated to clarify the impairment of B[a]P on ovaries at growing stage and mature stage. Overall, the present results elucidated the anti-estrogenic mechanisms along the steroidogenic pathway and estrogen signaling pathway for the stage-dependent endocrine-disrupting effects of B[a]P. This finding provides important information regarding to the underlying molecular mechanisms of B[a]P-induced endocrine disruption in different reproductive stages of bivalves. In addition, the adverse effects should be taken into concertation during protection of bivalves germplasm resources and comprehensive evaluation of ecological risks.

Chu kỳ sinh sản của điệp (giống Chlamys, họ Pectinidae) phân bố tại đảo Nam Du, tỉnh Kiên Giang

Scallops, belonging to Pectinidae family, were collected from Nam Du Island, Kien Giang province from September 2017 to August 2018 to study the reproductive biology, especially reproductive cycle. Gonadal somatic index (GSI) of scallops reached high values from September to November and from January to March, in which, the maximum value was recorded in November (8.29±2.51%) and January (8.91±2.69%). Female scallops presented high GSI from September to November, from January to February and in June, in which highest GSI occurred in January (12.48 ± 4.05%) and November (9.59 ± 4.22%). The positive relationship between GSI and egg volume of female scallops was observed in January, June, September and November. The results showed that scallops in Nam Du Island could spawn all year round; however, highest spawning synchronization occurred in January, June and November.

Seasonal variation of total carotenoids content in the tissues of male and female golden noble scallops Chlamys nobilis

The noble scallop Chlamys nobilis is an economically important edible marine bivalve that has been cultivated in the Southern Sea of China since the 1980s. Noble scallops, particularly the golden scallops, are rich in carotenoids, are of interest for their potential beneficial uses in human healthcare, food processing and pharmaceuticals. However, very little is known about the seasonal variation of total carotenoids content (TCC) in the golden scallops. Therefore, present study was conducted to determine the seasonal variation of TCC in the tissues of male and female of golden scallops. The results of present study revealed that the TCC in adductor, mantle and gonads of golden scallops were ranged from 16.79 to 138.86 μg/ g, 92.86 to 312.98 μg/ g, and 71.5 to 750.0 μg/ g, respectively. Generally, the gonads of golden scallops contain the highest TCC, followed by the mantle and adductor. In comparison of male and female scallops, TCC in the same tissue (except for gonad) was not statistically significant. However, female gonads contain much higher TCC than male gonads. The temporal variation of the TCC in golden scallops showed the highest in March to April and the lowest in October. The findings of current study can be very useful for scallop farmers and industry to determine the best harvest time to obtain the highest quality of golden scallops with high TCC.

Toxicological effects benzotriazole to the marine scallop Chlamys nobilis: a 2-month exposure study.

Benzotriazole and its associated derivatives (BTs) are widely used as ultraviolet stabilizers and corrosion inhibitors. They have been extensively found in marine environments and are bioaccumulative through the food chain. However, the toxicities of BTs to marine organisms are seldom identified and no assessment has been conducted for filter-feeding bivalves. In this study, a marine scallop Chlamys nobilis was exposed to 0, 0.01, 0.1, and 1.0mg/L of BT for 60days. Effects of BT on endocrine system, cytochrome P450 activity, antioxidant activity, and neural activity of C. nobilis were examined. The results showed that BT exerted significant estrogenic effects on both male and female scallops and inhibited EROD activities of C. nobilis even at 0.01mg/L level. BT at ≥ 0.01mg/L levels also caused significant oxidative stress on C. nobilis. Moreover, most of the adverse effects of BT to C. nobilis were found from day 35 and 0.01mg/L was the lowest concentration with observed effects, showing the long-term toxic effects of BT to C. nobilis. Thus, the adverse effects of BT and its derivatives to marine benthic communities deserve more attention in future research.

First evidence of marine diesel effects on biomarker responses in the Icelandic scallops, Chlamys islandica.

The decrease of ice cover in polar areas is expected to lead to an increase in ship traffic. In this context, the risk associated with exposure of the aquatic environment to oil-related chemicals from spills and/or accidental fuel discharges from ships will increase in the near future, potentially causing negative impacts on sensitive Arctic species. This study investigated for the first time the biomarker responses of the Icelandic scallops (Chlamys islandica) to marine diesel exposure. Antioxidant response, neurotoxic effect, gonad maturation, and lipid content were assessed in male and female scallops, Chlamys islandica after a 7-day exposure to marine diesel. At the end of exposure, results showed an inhibition of acetylcholinesterase activity in Icelandic scallops exposed to high concentration of diesel. At the end of exposure, results showed an increase in naphthalene residues in hemolymph, together with an inhibition of AChE activity in Icelandic scallops exposed to high concentration of diesel. In addition, alteration of the escape capacity was observed suggesting effects on behavior responses. Overall, this study contributes to the better knowledge of physiological effects of marine diesel in Arctic marine organisms.

Toxic effects upon exposure to polycyclic aromatic hydrocarbon (chrysene) in scallop Chlamys farreri during the reproduction period.

This study aims to investigate potential toxic effects of chrysene (CHR) on mature scallop Chlamys farreri during the reproduction period, using indicators of antioxidant defences and oxidative stress. Scallops were exposed to 0.2, 0.8 and 3.2μg/L waterborne CHR for 21 days, at day 10 scallops were induced to spawn. At days 1, 3, 6, 10, 11, 15 and 21, aryl hydrocarbon hydroxylase (AHH), 7-ethoxyresorufin-O-deethylase (EROD), glutathione-s-transferase (GST), glutathione (GSH), superoxide dismutase (SOD), lipid peroxidation (LPO), protein carbonyl (PC) and DNA strand breaks in digestive glands were examined by separately analysing male and female scallops. During the pre-spawn period, Levels of enzymatic activities and oxidative stress were all induced by the exposure to CHR for females and males. GST activity presented a good time- and dose-dependent relationship only in males, and GSH content showed a dose-dependent manner in both sexes. During the post-spawn period, different trends were observed, while PC contents maintained growth in time- and dose-dependent manner. Overall, males were more sensitive than females to CHR exposure in enzyme activities, and correspondingly, females suffered from more serious oxidative damages. Both GSH and PC contents seemed to be potential biomarkers for PAH exposure. These results will offer the information on toxicity of CHR in this species, and ensure the influence of gender and reproductive status on PAH detoxification metabolism.

Gender differences in detoxification metabolism of polycyclic aromatic hydrocarbon (chrysene) in scallop Chlamys farreri during the reproduction period

This study aims to investigate the effects of chrysene (CHR) on biotransformation and detoxification responses of mature scallop Chlamys farreri during the reproduction period. Scallops were exposed to 0.2, 0.8 and 3.2μg/L CHR for 21days; at day 10 scallops were induced to spawn. At days 1, 3, 6, 10, 11, 15 and 21, enzymatic activities of 7-ethoxyresorufin O-deethylase (EROD) and glutathione-s-transferase (GST), related mRNA expression levels of CYP1A1, GST-pi and P-glycoprotein (P-gp) in digestive glands and CHR bioaccumulation in tissues were examined by separately analyzing male and female scallops. During the pre-spawn period, CHR concentrations of the treated groups in tissues except the hemolymph increased rapidly. Levels of enzymatic activities and related gene expressions were all induced by the exposure to CHR for females and males. GST activity and GST-pi mRNA expression showed a good time- and dose-dependent relationship only in males, and P-gp mRNA expression exhibited a dose-dependent manner in both sexes. During the post-spawn period, spawning caused significant reductions of bioaccumulation in tissues but the gill and hemolymph. Enzymatic activities and related gene expressions were for females significantly depressed at day 21 at 0.8 or 3.2μg/L CHR. Overall, females accumulated more CHR than males, while males were more sensitive than females to CHR exposure in gene expressions and enzyme activities. P-gp mRNA expression seemed to be a potential biomarker for PAH exposure. These results will offer the information on CHR biotransformation in this species, and ensure the influence of gender and reproductive status on PAH detoxification metabolism.

An investigation of endocrine disrupting effects and toxic mechanisms modulated by benzo[a]pyrene in female scallop Chlamys farreri

The purpose of this study was to investigate the endocrine disrupting effects induced by benzo[a]pyrene (B[a]P) and explore the underlying mechanisms in mollusks. In this study, sexually mature female Chlamys farreri were exposed to benzo[a]pyrene for 10 days at four different concentrations as 0, 0.025, 0.5 and 10μg/L. Sex steroids were identified and quantified by electrochemiluminescence immunoassay (ECLIA) method and results showed that exposure to B[a]P exerts great suppression on 17β-estradiol, testosterone production and disrupts progesterone levels in ovary. Transcription of genes were detected and measured by real-time RT-PCR. It showed that at day 10 B[a]P inhibited 3β-HSD, CYP17 and 17β-HSD mRNA expression in a dose-dependent manner, which suggests that they could be potential targets of B[a]P that disrupt steroidogenic machinery. Moreover, 0.025μg/L B[a]P activated transcription of aryl hydrocarbon receptor (AHR), AHR nuclear translocator (ARNT), CYP1A1 and estrogen receptor (ER), while 10μg/L B[a]P suppressed all of them. The consistency of their responses to B[a]P exposure implies that AHR action may be involved in invertebrate CYP regulation and ER transcription despite of unknown mechanisms. Additionally, B[a]P exposure could induce ovarian impairment and developmental delay in C. farreri. Overall, sensitivity of C. farreri to endocrine disruption and toxicity suggests that C. farreri is a suitable species for study of endocrine-disrupting effects in marine invertebrates. This study will form a solid basis for a realistic extrapolation of endocrine disrupting effects across taxonomic groups and phyla.

The genomic structure, alternative splicing and immune response of Chlamys farreri thioester-containing protein

CfTEP is a member of thioester-containing protein (TEP) family found in Zhikong scallop Chlamys farreri and is involved in innate immunity against invading microbes. In the present study, the genomic DNA of CfTEP was cloned and characterized. The genomic DNA sequence of CfTEP consisted of 40 exons and 39 introns spanning 35 kb with all exon–intron junction sequences agreeing with the GT/AG consensus. The genomic organization of CfTEP was similar to human and mouse C3 rather than ciona C3-1 and Drosophila dTEP2. By RT-PCR technique, seven different cDNA variants of CfTEP (designated as CfTEP-A–CfTEP-G) were cloned from scallop gonad. CfTEP-A–CfTEP-F were produced by alternative splicing of six mutually exclusive exons (exons 19–24), respectively, which encoded the highly variable central region. While in CfTEP-G, the deletion of all the six exons introduced a new translation stop site and might trigger nonsense mediated decay (NMD). The mRNA expression and the proportion of the seven CfTEP variant transcripts were examined in the gonad of scallops after bacterial challenge. The fragments containing the highly variable central region of CfTEP were amplified by RT-PCR and a 100 positive clones were sequenced randomly. The expression profiles of the seven CfTEP variants were different and displayed the sex and bacteria dependent manner. In the blank, sea water and Listonella anguillarum challenged subgroups of male scallops, all the transcripts detected were CfTEP-G isoform. In the Micrococcus luteus challenged subgroup, the isoforms expressed and their proportions were CfTEP-F (54%), CfTEP-B (23%), CfTEP-A (10%), CfTEP-C (7%) and CfTEP-E (6%). However, in the gonad of female scallops, only CfTEP-A were found in blank and sea water challenged subgroups. After L. anguillarum or M. luteus challenge, four and five isoforms were detected, respectively, with CfTEP-F isoform being the most one in the both subgroups. These results suggested that the evolution of TEP genes was very complex, and that the diverse CfTEP transcripts generated by alternative splicing played an important role as pattern recognition receptors in the innate immune defense of scallops.

Uptake and effects of copper and cadmium in the gonad of the scallop Placopecten magellanicus: concurrent metal exposure

Sea scallops Placopecten magellanicus in early gametogenesis from the southern shelf of Hudson Canyon, New Jersey, USA, were exposed in late winter 1984 to sublethal levels of Cu and Cd in a flowing seawater system at the NMFS Milford Laboratory. Exposure was to copper (10 and 20 μgl-1: low-Cu and high-Cu groups) or to a combination of copper and equimolar cadmium (10μg Cu+17.7 μg Cdl-1: low-Cu/Cd group) for eight weeks, with sampling at 2-wk intervals. Copper had a strongly inhibitory effect on gamete production and maturation, which in some respects was partially moderated in the presence of cadmium in the female gonad only. Total gamete weight per scallop doubled in control individuals but dropped by 60% in both high-metal exposure groups over the 8-wk exposure period, with a smaller, temporary decrease in the low-Cu group. Cadmium did not add to the inhibition by copper of gamete development in the low-Cu/Cd group, but there was no partial recovery at 8 wk, as was seen in female scallops exposed to low-Cu alone. Gonadal RNA, higher in the females, decreased proportionately more in that sex than in the males of the metal exposure groups. Conversely, DNA levels were higher in the male than in the female gonad, and decreased sharply in all metal-exposed males. Gonadal protein concentration also dropped in all metal-exposed scallops with time and degree of metal exposure. Copper uptake in the gonad increased with time and metal exposure concentration, and cadmium increased in the low-Cu/Cd group with time. In contrast, manganese decreased significantly in the gonads of Cu-exposed scallops, especially in the high-Cu group. In the low-Cu/Cd group, manganese concentrations stabilized after an initial sharp drop at 2 wk in the low-Cu/Cd group, then rose at 6 and 8 wk. We ascribe this phenomenon to the induction by cadmium of metal-binding proteins (Fowler and Megginson 1986). In both sexes, gonadal magnesium concentrations did not change with increasing tissue burdens of copper and cadmium, but instead rose initially in proportion to the degree of metal exposure, homeostasis being maintained thereafter.