Lead and Aroclor 1254 disrupt reproductive neuroendocrine function in Atlantic croaker

We have previously shown that lead (lead chloride) and a polychlorinated biphenyl (PCB) mixture (Aroclor 1254) can alter hypothalamic serotonin (5-hydroxytryptamine, 5-HT) content, and pituitary gonadotropin II (GTH II) release in vitro, in the Atlantic croaker (Micropogonias undulatus). In the present study we investigated whether impairment of the hypothalamic 5-HT pathway involves alterations in tryptophan hydroxylase (TPH), the rate-limiting enzyme in 5-HT biosynthesis, or monoamine oxidase (MAO), the catabolic enzyme. Aroclor 1254 (1 mg/kg body wt. for 30 days) significantly inhibited hypothalamic TPH activity without altering MAO activity, and caused a significant decline in 5-HT content. On the other hand, lead exposure (15 mg/kg body wt. for 30 days) only induced a slight decrease in hypothalamic 5-HT content and TPH activity, and a minor increase in MAO activity. However, both Aroclor 1254 and lead significantly inhibited the GTH II response to stimulation by a luteinizing hormone-releasing hormone analog (LHRHa) in vivo and caused reduced gonadal growth. These results demonstrate that impairment of hypothalamic serotonin metabolism by Aroclor 1254 involves inhibition of 5-HT synthesis, whereas lead does not exert a profound influence on 5-HT metabolism. The decline in 5-HT availability due to reduced 5-HT synthesis in the PCB-exposed fish may result in disruption of the stimulatory 5-HT-GnRH pathway controlling GTH II secretion leading to impairment of gonadal growth.     

Vitamin E co-treatment reduces Aroclor 1254-induced impairment of reproductive neuroendocrine function in Atlantic croaker

We have shown previously that exposure of Atlantic croaker to a PCB mixture (Aroclor 1254) results in impaired reproductive neuroendocrine function. In addition, we have identified hypothalamic tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin (5-hydroxytryptamine) synthesis, as a target of PCB neuroendocrine toxicity. In order to further elucidate the mechanisms of PCB neurotoxicity, the present study investigated whether PCB-induced decrease in hypothalamic TPH activity resulted from degradation of the enzyme protein. Fish were exposed to Aroclor 1254 in the diet (0.1 mg/100 g body weight (BW)/day) for 30 days. The PCB exposure elicited a significant decrease in hypothalamic TPH protein content, which could be at least partially responsible for the reduced TPH activity. To test whether a similar PCB exposure could cause oxidative damage in croaker hypothalamic tissues, we examined the formation of malondialdehyde (MDA) protein adducts as a marker of lipid peroxidation (LPO). The same dose of PCB increased the MDA-protein adduct formation in the hypothalamus. In a separate experiment, the role of vitamin E, an antioxidant, to prevent or decrease the effect of PCB on hypothalamic TPH activity and gonadal growth was examined. The vitamin E co-treatments (1 and 10 mg/100 g BW/day) with PCB significantly reduced the effects of PCB on TPH activity and gonadal growth. These results suggest possible involvement of oxidative processes in PCB neurotoxicity.     

细胞外钙和钾离子对调节离体鲤脑垂体促性腺激素分泌的影响

作者于１９９０年９月－１９９１年１月采用离体灌流培育方法对鲤鱼脑垂体促性激素的分泌调节进行研究。结果表明，脑垂体基础促性腺激素（ＧｔＨ）分泌不依赖于细胞外钙离子（Ｃａ＾２＋）；但脉冲式鲑鱼促性腺激素释放激素（ｓＧｎＲＨ）刺激的ＧｔＨ分泌，则依赖于细胞外Ｃａ＾２＋且细胞外Ｃａ＾２＋的作用至少部分通过电位敏感性Ｃａ＾２＋通道。５０ｍｍｏｌ／Ｌ高浓度钾离子（Ｋ＾＋）显著刺激脑垂体基础ＧｔＨ分泌，Ｋ＾＋的     

Effects of Aroclor 1254 on reproductive organ weight, sperm quality and testicular mitochondria oxidative stress of Boleophthalmus pectinirostris

Boleophthalmus pectinirostris is an amphibious economic fish and wildly distributed in the southeast coast of China. In this study, Aroclor 1254 was intraperitoneally injected into B. pectinirostris with 1, 2 and 4 μg/(g·d) for 28 d to assay the reproductive organ weight, the sperm quality (sperm concentration and motility), and the testicular mitochondrial testicular mitochondria oxidative stress. The results show that the sperm number and motility in seminal vesicles, the absolute weight of testes and seminal vesicles of B. pectinirostris treated with 2 and 4 μg/(g·d) Aroclor 1254 decreased significantly as compared to the controls (p <0.05), while those treated with 1 μg/(g·d) Aroclor 1254 had no significant effects on these indictors. For the relative weight of reproductive organs, significant reduction (p <0.05) was only observed in the seminal vesicles of B. pectinirostris treated with 4 μg/(g·d). SOD activities and GSH levels in all the Aroclor 1254 treatments were significantly lower than those of the controls (p <0.05). The activities of CAT, GPx, GR and the levels of Vit C also decreased significantly in comparison with the controls (p <0.05) at the higher dose of 2 and 4 μg/(g·d) Aroclor 1254 treatments. In addition, both H₂O₂ level and MDA content in testicular mitochondria of B. pectinirostris had a close correlation with Aroclor 1254 dosage, and were significantly higher than the controls (p <0.05). Those indicate that Aroclor 1254 can induce the oxidative stress of testicular mitochondria, and impair the reproductive function of male B. pectinirostris.     

Regulation of hepatic glutathione S-transferase expression in flounder

The major glutathione S-transferase isoform of flounder liver, an antigenically related structural homologue of plaice GST-A, also displays mRNA homology. A 901bp cRNA probe for plaice GST-A cross-hybridised to a 1100bp flounder mRNA on northern blot analysis. The plaice antibody and cRNA probes were used to study effects of inducer treatment on GST-A expression in flounder liver. Six days after PAH treatment (3-methylcholanthrene) total hepatic GST activity was halved, levels of GST-A were 80% and GST-A mRNA levels were 25% of controls. A commercial PCB mixture (Aroclor 1254^TM) had little effect on total GST or GST-A levels despite halving GST-A mRNA levels. An epoxide, trans-stilbene oxide induced total GST activity 1·4 fold and GST-A protein levels 1·8-fold and its mRNA levels 3-fold. This reduced expression of the major flounder hepatic GST by agents which induce cytochrome P4501A1 may modulate cytoxicity of environmental pollutants in this species.     

PCB uptake and accumulation by oysters (Crassostrea virginica) exposed via a contaminated algal diet

Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 μg polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills+mantle and muscle of oysters fed algal paste containing 1.0 μg PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills+mantle tissues compared to the same tissues in the undosed control.     

PCB uptake and accumulation by oysters (Crassostrea virginica) exposed via a contaminated algal diet.

Reproductively active oysters were fed daily with 0.2 g algal paste containing 0, 0.1, and 1.0 microgram polychlorinated biphenyls (PCBs) (1:1:1 mixture of Aroclor 1242, 1254 and 1260) for either 15 or 30 days, and accumulation of PCBs in different organ tissues and eggs assessed. The effects of PCB exposure on lipid content, lipid class and fatty acid composition were also evaluated. PCBs were accumulated by the oysters and transferred to the eggs. PCB accumulation in oysters was dose, time and tissue dependent. Mean PCB contents were 3150, 1970, and 250 ng/g dry wt., respectively, in the visceral mass, gills + mantle and muscle of oysters fed algal paste containing 1.0 microgram PCBs for 30 days. The PCBs in the eggs from the same oysters reached 671 ng PCBs/g dry wt. Feeding oysters with PCB-sorbed algal paste for 30 days significantly increased phospholipid and free fatty acid contents in gills + mantle tissue compared to the same tissues in the undosed control.     

海湾扇贝幼虫变态过程中体内神经递质含量的变化

采用生化方法测定了不同发育阶段及人工诱导后海湾扇贝 (Argopectenirradians)幼虫体内去甲肾上腺素 (NE)、多巴胺 (DA)和 5 羟色胺 ( 5 HT)含量的变化。结果表明 ,海湾扇贝幼虫体内去甲肾上腺素含量在变态前后没有明显变化 ,变态前为 2 35 2pg/mg湿重 ,变态后为2 770pg/mg湿重。多巴胺和 5 羟色胺含量在变态前 (第 1 3天 )急剧增加 ,比第 1 2天的幼虫分别增加了 2 .8倍和 4.7倍 ,变态后急剧下降 ,变态后幼苗比第 1 3天的幼虫分别降低了 2 5 .1倍和 1 6.4倍。海湾扇贝幼虫体内DA∶NE比值和 5 HT∶NE比值在变态前后变化剧烈。DA∶NE比值和 5 HT∶NE比值在变态前 (第 1 3天 )急剧增加 ,比第 1 2天的幼虫增加了 3.0倍 (DA∶NE)和 5 .0倍 ( 5 HT∶NE) ;变态后急剧降低 ,变态后幼苗比第 1 3天的幼虫降低了 2 9.8倍 (DA∶NE)和 1 9.5倍 ( 5 HT∶NE)。海湾扇贝幼虫经氯化钾和氯化钙诱导 2 4h后 ,体内去甲肾上腺素、多巴胺和 5 羟色胺以及DA∶NE比值和 5 HT∶NE比值均有所降低。本实验的结果表明 ,多巴胺和 5 羟色胺可能启动了海湾扇贝幼虫的变态过程。     

Resistance may be an important mechanism by which marine microbes respond to environmental toxicants

The hypothesis, that at least certain marine microbial organisms respond to toxic stress by the development of resistance, was tested using the hypotric marine ciliate Euplotes vannus Muller as the test organism. Resistance to polychlorinated biphenyls (PCB, Aroclor 1254) was developed in E. vannus by exposing the animals to progressively higher PCB concentrations during a period of several months. The resistance to PCB persisted for at least 80 days (greater than 40 generations) after final exposure. This suggests either that genetic selection or persistent (lasting over many cell division cycles) phenotypic trait modification, possibly in the form of Dauermodification, had occurred. If resistance were widespread among marine microbial organisms in polluted environments it would be an important consideration in evaluating the long-term biological impact of both natural and man-induced chemical stress.     

Factors affecting the elimination of PCBs in the marine copepod Acartia tonsa

The effects of feeding, egg laying, and fecal pellet production on the elimination of polychlorinated biphenyls (PCBs) from the marine copepod Acartia tonsa were studied in a series of experiments. Copepods were exposed to ¹⁴C-labelled Aroclor 1254 and allowed to depurate in clean seawater. Copepods fed during depuration eliminated PCBs more rapidly than unfed copepods whether or not the original PCB exposure medium had contained food. Both eggs and fecal pellets contained PCBs during depuration, with the weight specific concentration of PCB in the eggs (up to 407 ppm, dry weight) exceeding four times that in the females that produced them. Female copepods eliminated PCBs twice as rapidly as males, indicating that egg production is an important route for PCB elimination.     

神经递质多巴胺(DA)和5-羟色胺(5-HT)对三种主要养殖对虾争胜行为的影响

本文采用肌肉注射和行为分析的方法,进行了注射多巴胺(DA)和5-羟色胺(5-HT)对对虾争胜行为的研究,比较分析了注射多巴胺(DA)和5-羟色胺(5-HT)对日本囊对虾(Penaeus japonicus)、凡纳滨对虾(Litopenaeus vannamei)和中国明对虾(Fenneropenaeus chinensis)争胜行为的影响。多巴胺注射浓度为2×10~(-3)mol/L、2×10~(-4)mol/L,5-羟色胺注射浓度为2×10~(–5)mol/L、2×10~(–6)mol/L,以生理盐水为对照,各设3个平行组。检测注射1h后对虾体内的DA、5-HT含量,分析比较注射DA、5-HT前后及不同对虾种类争胜行为的差异。结果表明,注射多巴胺后三种对虾间的争斗显著增加,HDA比LDA更易激发对虾间的争斗,同种对虾在注射不同浓度多巴胺后与对照组差异均显著(P0.05);相较于对照组,南美白对虾、日本囊对虾、中国明对虾在注射低浓度多巴胺后,对虾平均争斗次数分别增加13.8%、11.1%、21.8%,高浓度多巴胺组分别增加91.0%、64.0%、68.5%;对虾平均争斗次数随着DA浓度的升高而增加;平均优势指数总体呈下降趋势。随着5-HT浓度的增加,对虾的平均争斗次数随之减少,相较于对照组,低浓度5-羟色胺组降幅超过50%,高浓度5-羟色胺组降幅超过270%以上。争斗次数表现为中国明对虾凡纳滨对虾日本囊对虾。平均优势指数随注射5-HT浓度的增加而小幅度上升。     

外源神经递质5-羟色胺(5-HT)和多巴胺(DA)调控三疣梭子蟹争斗行为的初步研究

影响甲壳类动物争斗行为的因素包括性别差异、体型和生殖状态等,神经递质对甲壳动物的争斗行为也具有调控作用。为探索三疣梭子蟹在养殖和繁殖过程中由于争斗行为受损或死亡的行为机制,采用双抗体夹心法测定争斗前后三疣梭子蟹血淋巴中5-HT和DA的含量,并分析了争斗前后肌肉、胸神经节、肝胰腺三种组织中5-HT和DA受体基因表达量变化。同时,通过注射外源5-HT和DA,记录并分析了三疣梭子蟹争斗行为差异以及三种组织中5-HT和DA受体基因表达量变化。结果表明:(1)争斗后三疣梭子蟹血淋巴中5-HT含量显著升高,DA含量显著降低(P<0.05);(2)注射10^–6 mol5-HT能显著减少三疣梭子蟹的争斗行为发生次数以及争斗持续时间(P<0.05),而注射10^–6 molDA却能够显著增加三疣梭子蟹的争斗行为发生次数和争斗持续时间(P<0.05);(3)争斗前后5-HT和DA受体在肌肉和胸神经节中表达量发生显著变化(P<0.05),且不同受体在相同组织中未呈现相同的变化趋势;(4)注射外源DA能够调节5-HT1B、5-HT2B、5-H...     

The use of polyclonal antibodies raised against rat and trout cytochrome P450 CYP1A1 orthologues to monitor environmental induction in the channel catfish (Ictalurus punctatus)

Induction of hepatic cytochrome P450-dependent microsomal mono-oxygenase by xenobiotics is a well-established phenomenon in teleost fish. As in laboratory mammals, fish possess multiple forms of cytochrome P450 that display overlapping substrate specificity. One such isoform, CYP1A1, which has been cloned and sequenced from rainbow trout, has been shown to be orthologous to rat CYP1A1 and, as in mammals, is inducible up to several hundred-fold by planar aromatic hydrocarbons, PCBs and dioxins. It has been suggested that induction of CYP1A1 orthologues might provide a sensitive biomonitor for environmental pollution by mixtures of such compounds. In the current study, polyclonal antibodies directed against CYP1A1 purified from rat and trout liver were used to monitor induction of the CYP1A1 orthologue in hepatic microsomes from the fresh water species, the channel catfish (Ictalurus punctatus). Catfish from a local fish farm were induced in the laboratory by three daily injections of 50 mg/kg of the PCB mixture Aroclor 1254 and compared with fish taken from a site in central Arkansas—the Bayou Meto, known to be polluted with dioxin. Hepatic microsomal activities towards ethoxyresorufin (EROD) and pentoxyresorufin (PROD) were measured and Western blot analysis carried out with the two antibodies. EROD was elevated in both the Aroclor-treated fish and in the Bayou Meto fish compared with untreated fish farm controls; smaller but significant increases were observed in PROD. Spearman's rank correlations of 0·74 and 0·89 were observed between EROD and immunoquantified cross-reactivity towards the rat CYP1A1 and trout CYP1A1 antibodies.     

Metabolism of perylene by monooxygenases of the skin and liver of Ambystoma tigrinum

The prevalence of dermal neoplasia among neotenic tiger salamanders (Ambystoma tigrinum) collected from the Reese Air Force Base sewage lagoon far exceeded that among salamanders from nearby uncontaminated lagoons. Perylene was the predominant pglycyclic aromatic hydrocarbon contaminant in this lagoon. Perylene was shown to be a substrate for monooxygenases in skin and liver homogenates and formed DNA-binding metabolites. Perylene exposure appeared to augment subsequent perylene metabolism by dermal enzymes but it was not a strong inducer of hepatic monooxygenases. Aroclor 1254 induced benzo(a)pyrene and perylene metabolism by both skin and liver enzymes. However, perylene apparently was not activated by salamander enzymes, as suggested by its lack of in vitro mutagenicity and failure to induce tumors.     

Behavior and brain chemistry correlates in mummichogs (Fundulus heteroclitus) from polluted and unpolluted environments

The ability of Fundulus from a polluted environment to capture a prey organism, the grass shrimp (Palaemonetes pugio), was compared to that of Fundulus from an unpolluted reference site. Fish from the unpolluted site took significantly more prey (p < 0.1) during the first 5–10 min of each trial than fish from the polluted site.Analysis of the neurotransmitters dopamine (DA) and serotonin (5-HT) and their metabolites, DOPAC, homovanillic acid (HVA), and 5-hydroxy-indolacetic acid (5-HIAA) in cerebellums and medullas was performed by HPLC. Mummichogs from the polluted site had significantly lower concentrations of 5-HT and its metabolite 5-HIAA in their medullas, but not cerebellums, than fish from the reference site (p < 0.05). Concentrations of DA, DOPAC and HVA were not different between populations in either part of the brain. Neurotransmitter concentrations were shown to be independent of size and therefore, presumably, age.Analysis of Hg in livers and brains of these mummichogs by atomic absorption spectrophotometry showed significant differences between the two populations for both organs. Earlier work showed that the behavioral deficit could be elicited by Hg; thus, this was the putative neurotoxic agent. However, there are other candidates which may contribute to neurotoxicity in the study site.     

多氯联苯(PCB1254)对栉孔扇贝消化盲囊和鳃丝抗氧化系统及脂质过氧化水平的影响

本实验研究了浓度梯度为0.5,1.0,和10.0μg/L的多氯联苯化舍物(PCB1254)处理30d栉孔扇贝消化盲囊和鳃丝SOD,CAT和GPx活性和脂质过氧化水平的变化.研究结果表明:低浓度组(0.5μg/L和1.0μg/L)的抗氧化酶活力能够被PCB1254显著的诱导,但是在高浓度组,抗氧化酶快速被诱导升高然后随着...     

Transport and accumulation of organochlorines in the ovaries of Atlantic croaker (Micropogonias undulatus)

The role of vitellogenin in the transport of organochlorines was investigated in Atlantic croaker (Micropogonias undulatus) by exposing them through the diet to o,p′-DDT at a concentration of 10.8 μg/100 g fish/day for 14 days or Aroclor 1254 (PCB) at a concentration of 0.5 mg/100 g fish/day for 30 days during gonadal recrudescence. Tissue samples were taken from the fish at various times after initial exposure, and o,p′-DDT and PCBs were extracted in acetonitrile and analyzed by gas chromatography. Analysis of the ovarian tissue collected 2 and 3 weeks from the start of exposure revealed that the o,p′-DDT concentration increases as the gonadosomatic index (GSI) increases (r² = 0.63), with accumulation ranging from less than 1% to as much as 8% of the total dosage. Interestingly, o,p′-DDT did not accumulate in the testes during the same exposure period. Accumulation of PCBs was found to be 40 times higher in the ovaries than in the testes. Gel filtration of plasma from exposed females showed that o,p′-DDT elutes in the low density lipoprotein and vitellogenin fractions. Control plasma incubated with o,p′-DDT at 4 °C for 16 h followed by chromatography on Sepharose 6B gave similar results with an o,p′-DDT concentration of 0.6μg/mg protein in the vitellogenin fraction. Furthermore, both o,p′-DDT and PCBs were found to bind to purified croaker vitellogenin. These results suggest that lipoproteins, including vitellogenin, are involved in the transport and accumulation of organochlorines in the ovaries of exposed fish.     

皱瘤海鞘神经递质和性类固醇激素含量与性腺发育成熟的关系

运用免疫组织化学和化学发光免疫分析方法研究了皱瘤海鞘(Styela plicata)神经递质多巴胺(DA)、5-羟色胺(5-HT)及其受体以及性类固醇激素含量与性腺发育成熟的关系.结果显示,DA和5-HT免疫活性定位在脑神经节中大小2种类型的神经细胞,DA免疫活性还定位在不同发育时期神经腺上皮细胞.5-HT受体免疫活性存在于脑神经节小型神经细胞和大生长期神经腺上皮细胞.在小生长期和大生长期性腺中均检测到DA、5-HT及其受体,提示DA和5-HT可能参与调节皱瘤海鞘的性腺发育.皱瘤海鞘为雌雄同体,化学发光免疫分析证实,海鞘性腺能够合成和分泌性类固醇激素,大生长期性腺中雄激素、孕激素和雌激素的含量分别为25.8±0.0、38.9±0.2和14.3±0.6pg/mg,提示性腺既是生殖组织也是内分泌组织.上述发现为证明尾索类海鞘可能像头索类文昌鱼一样,具有原始的生殖内分泌调控轴(脑神经节-神经腺-性腺)提供科学依据.     

Reproductive endocrine function in female atlantic croaker exposed to pollutants

To examine whether xenobiotics impair teleost reproduction by altering reproductive endocrine function, steroid hormone secretion and ovarian growth were investigated in female Atlantic croaker (Micropogonias undulatus) after chronic oral administration with sublethal doses of three classes of reproductive toxins (lead, benzo[a]pyrene and Aroclor 1254). All the sublethal treatments significantly impaired ovarian growth as assessed by the gonadosomatic index. Reduced ovarian growth was accompanied by a significant decline in circulating 17β-estradiol levels in fish exposed to lead and benzo[a]pyrene. All three toxicants significantly decreased plasma testosterone levels. However, the steroidogenic capacity of ovarian tissue in vitro was not altered by xenobiotic exposure. The data suggest that the decreased ovarian growth in croaker after pollutant exposure may be a consequence of the decline in plasma 17β-estradiol levels. Further, this decrease in circulating levels of gonadal steroids does not appear to be caused by a direct effect of the chemicals on ovarian steroid synthesis.