This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of t...This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.展开更多
This study examined the effect of salinity on the expression of Na+/K+-ATPase(NKA) α-subunit and vacuolar-type H+-ATPase(V-ATPase) β-subunit gene in the gill of Litopenaeus vannamei. Semi-quantitative reverse transc...This study examined the effect of salinity on the expression of Na+/K+-ATPase(NKA) α-subunit and vacuolar-type H+-ATPase(V-ATPase) β-subunit gene in the gill of Litopenaeus vannamei. Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) assay showed that the expression of NKA α-subunit and V-ATPase β-subunit gene was significantly influenced by salinity. It was found that the NKA activity significantly varied with salinity in time and dose dependent manner; whereas the V-ATPase activity did not. The abundance of NKA α-subunit gene transcript increased rapidly when the salinity decreased from 26 b to 21, and slowly when the salinity decreased from 26 to 31 within the first 24 h. When the salinity decreased from 26 to 21, the transcription of NKA α-subunit gene in gill epithelium was higher at 12 h than that at 0 h, which was consistent with the result of immunoblotting assay of NKA α-subunit. In addition, salinity had a significant time- and dose-dependent effect on the concentration of biogenic amines in both hemolymph and gill. As compared to other parameters, the concentration of dopamine(DA) and 5-hydroxytryptamine(5-HT) varied in different patterns when the salinity decreased from 26 to 21 or increased from 26 to 31, suggesting that DA and 5-HT played different regulatory roles in osmotic adaption and modulation of shrimp when salinity varies.展开更多
The objective of this study was to examine the effect ofbenzo[a]pyrene （BaP） on the detoxification and antioxidant systems of two microalgae, Isochrysis zhanjiangensis and Platymonas subcordiformis. In our study, th...The objective of this study was to examine the effect ofbenzo[a]pyrene （BaP） on the detoxification and antioxidant systems of two microalgae, Isochrysis zhanjiangensis and Platymonas subcordiformis. In our study, these two algae were exposed to BaP for 4 days at three different concentrations including 0.5 μg L-1 （low）, 3 μg L-1 （mid） and 18 μg L-1 （high）. The activity of detoxi- fication enzymes, ethoxyresorufin O-deethylase （EROD） and glutathione S-transferase （GST） increased in P subcordiformis in all BaP-treated groups. In 1. zhanjiangensis, the activity of these two enzymes increased at the beginning of exposure, and then de- creased in the groups treated with mid- and high BaP. The activity of antioxidant enzyme superoxide dismutase （SOD） increased in/. zhanjiangensis in all BaP-treated groups, and then decreased in high BaP-treated group, while no significant change was observed in P subcordiformis. The activity of antioxidant enzyme catalase （CAT） increased in I. zhanjiangensis and P subcordiformis in all BaP- treated groups. The content of malondialdehyde （MDA） in Isochrysis zhanjiangensis increased first, and then decreased in high BaP-treated group, while no change occurred in P. subcordiformis. These results demonstrated that BaP significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae. The metabolic related enzymes （EROD, GST and CAT） may serve as sensitive biomarkers of measuring the contamination level of BaP in marine water.展开更多
The Pacific white shrimp, Litopenaeus vannamei, is widely farmed in China. Salinity is a major environmental factor that affects its growth and distribution. Crustacean hyperglycemic hormone is verified to participate...The Pacific white shrimp, Litopenaeus vannamei, is widely farmed in China. Salinity is a major environmental factor that affects its growth and distribution. Crustacean hyperglycemic hormone is verified to participate in ion transport in response to the salinity challenge mediated by endocrine neurotransmitters(biogenic amines, BAs). In the present study, the contents of BAs and expressions of their receptors were detected in gills of Litopenaeus vannamei exposed to low salinity. The intracellular signaling molecules such as cyclic adenosine monophosphate(cAMP), protein kinase A(PKA), 14-3-3 protein, FXYD2 protein and cAMP response element-binding protein(CREB) were detected. The effects of low salinity on the expressions of Na^+-K^+-ATPase, Na^+/K^+/2 Cl-co-transporter and Cl-transporter and activity of Na^+-K^+-ATPase were also analyzed. The results showed that dopamine and epinephrine concentrations and their receptor expressions were significantly affected by low salinity. The changes of c AMP and PKA were obvious and the expressions of 14-3-3 and FXYD2 peaked at early stages. However, the expression of CREB was only significantly up-regulated on day 9. The activity and expression of Na^+-K^+-ATPase(α subunit) reached a peak on day 1. The expressions of Na^+/K^+/2 Cl-co-transporter and Cl-transporter up-regulated obviously. It suggests that BAs can activate the cAMP-PKA pathway, which further acts on the 14-3-3 and FXYD2 proteins, and ultimately improve the activity of Na^+-K^+-ATPase. Furthermore, after BAs stimulate the cAMP-PKA pathway, PKA phosphorylates the transcription factor CREB and regulates the expressions of ion transport enzymes/transporters. The results in this study are helpful for understanding the response mechanism of endocrine neurotransmitters on osmoregulation in crustaceans.展开更多
The ovary is an excellent system for studying stem cell renewal and differentiation, which is under the control of ovarian somatic cells. In order to understand oogenesis in Fugu rubripes （Temminck et. Schlegel） as ...The ovary is an excellent system for studying stem cell renewal and differentiation, which is under the control of ovarian somatic cells. In order to understand oogenesis in Fugu rubripes （Temminck et. Schlegel） as a marine fish model of aquaculture importance, we established cell lines called TSOC 1 and TSOC2 from a juvenile ovary of this organism. TSOC1 is composed of spindle epithelial-like cells, while the other is cobblestone-like ceils. Therefore, TSOC1 and TSOC2 appear to consist of ovarian somatic cells. Growth requirement condition was investigated including temperature, concentration of FBS and pH. Significant fluorescent signals were observed after TSOC1 and TSOC2 cells were transfected with pEGFP-N3 vector, indicating its potential utility for genetic manipulation such as gene function studies. It is shown that these cell lines are effective for infection by the turbot reddish body iridovirus and flounder lymphosystis disease virus as evidenced by the appearance of cytopathic effect and virus propagation in the virus-infected cells, and most convincingly, the observation of viral particles by electron microscopy, demonstrating that TSOC1 and TSOC2 are suitable to study interactions between virus and host cells. It is believed that TSOC1 and TSOC2 will be useful tools to study sex-related events and interactions between primordial germ cells and oogonia cells during oogenesis. Therefore, establishment of ovary cell lines from Fugu rubripes seems to be significant for those research areas.展开更多
基金supported by the Natural Science Foundation of Shandong Province, China (No. ZR2016 CM21)
文摘This study investigated the effects of ammonia-N(control and 1 and 5 mg L^(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.
基金supported by the National Natural Sciences Foundation of China (31072193)the Scientific Research Foundation for Outstanding Young Scientists of Shandong Province (2006BS07005)
文摘This study examined the effect of salinity on the expression of Na+/K+-ATPase(NKA) α-subunit and vacuolar-type H+-ATPase(V-ATPase) β-subunit gene in the gill of Litopenaeus vannamei. Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) assay showed that the expression of NKA α-subunit and V-ATPase β-subunit gene was significantly influenced by salinity. It was found that the NKA activity significantly varied with salinity in time and dose dependent manner; whereas the V-ATPase activity did not. The abundance of NKA α-subunit gene transcript increased rapidly when the salinity decreased from 26 b to 21, and slowly when the salinity decreased from 26 to 31 within the first 24 h. When the salinity decreased from 26 to 21, the transcription of NKA α-subunit gene in gill epithelium was higher at 12 h than that at 0 h, which was consistent with the result of immunoblotting assay of NKA α-subunit. In addition, salinity had a significant time- and dose-dependent effect on the concentration of biogenic amines in both hemolymph and gill. As compared to other parameters, the concentration of dopamine(DA) and 5-hydroxytryptamine(5-HT) varied in different patterns when the salinity decreased from 26 to 21 or increased from 26 to 31, suggesting that DA and 5-HT played different regulatory roles in osmotic adaption and modulation of shrimp when salinity varies.
基金supported by the State Oceanic Administration Specific Public Project of China (201105013)
文摘The objective of this study was to examine the effect ofbenzo[a]pyrene （BaP） on the detoxification and antioxidant systems of two microalgae, Isochrysis zhanjiangensis and Platymonas subcordiformis. In our study, these two algae were exposed to BaP for 4 days at three different concentrations including 0.5 μg L-1 （low）, 3 μg L-1 （mid） and 18 μg L-1 （high）. The activity of detoxi- fication enzymes, ethoxyresorufin O-deethylase （EROD） and glutathione S-transferase （GST） increased in P subcordiformis in all BaP-treated groups. In 1. zhanjiangensis, the activity of these two enzymes increased at the beginning of exposure, and then de- creased in the groups treated with mid- and high BaP. The activity of antioxidant enzyme superoxide dismutase （SOD） increased in/. zhanjiangensis in all BaP-treated groups, and then decreased in high BaP-treated group, while no significant change was observed in P subcordiformis. The activity of antioxidant enzyme catalase （CAT） increased in I. zhanjiangensis and P subcordiformis in all BaP- treated groups. The content of malondialdehyde （MDA） in Isochrysis zhanjiangensis increased first, and then decreased in high BaP-treated group, while no change occurred in P. subcordiformis. These results demonstrated that BaP significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae. The metabolic related enzymes （EROD, GST and CAT） may serve as sensitive biomarkers of measuring the contamination level of BaP in marine water.
基金supported by State Oceanic Administration Specific Public Project of China (No. 201305005)the National Natural Science Foundation of China (No. 31072193)
文摘The Pacific white shrimp, Litopenaeus vannamei, is widely farmed in China. Salinity is a major environmental factor that affects its growth and distribution. Crustacean hyperglycemic hormone is verified to participate in ion transport in response to the salinity challenge mediated by endocrine neurotransmitters(biogenic amines, BAs). In the present study, the contents of BAs and expressions of their receptors were detected in gills of Litopenaeus vannamei exposed to low salinity. The intracellular signaling molecules such as cyclic adenosine monophosphate(cAMP), protein kinase A(PKA), 14-3-3 protein, FXYD2 protein and cAMP response element-binding protein(CREB) were detected. The effects of low salinity on the expressions of Na^+-K^+-ATPase, Na^+/K^+/2 Cl-co-transporter and Cl-transporter and activity of Na^+-K^+-ATPase were also analyzed. The results showed that dopamine and epinephrine concentrations and their receptor expressions were significantly affected by low salinity. The changes of c AMP and PKA were obvious and the expressions of 14-3-3 and FXYD2 peaked at early stages. However, the expression of CREB was only significantly up-regulated on day 9. The activity and expression of Na^+-K^+-ATPase(α subunit) reached a peak on day 1. The expressions of Na^+/K^+/2 Cl-co-transporter and Cl-transporter up-regulated obviously. It suggests that BAs can activate the cAMP-PKA pathway, which further acts on the 14-3-3 and FXYD2 proteins, and ultimately improve the activity of Na^+-K^+-ATPase. Furthermore, after BAs stimulate the cAMP-PKA pathway, PKA phosphorylates the transcription factor CREB and regulates the expressions of ion transport enzymes/transporters. The results in this study are helpful for understanding the response mechanism of endocrine neurotransmitters on osmoregulation in crustaceans.
基金The finance special program of Tianjinthe transformation project of Tianjin Agricultural Achievements
文摘The ovary is an excellent system for studying stem cell renewal and differentiation, which is under the control of ovarian somatic cells. In order to understand oogenesis in Fugu rubripes （Temminck et. Schlegel） as a marine fish model of aquaculture importance, we established cell lines called TSOC 1 and TSOC2 from a juvenile ovary of this organism. TSOC1 is composed of spindle epithelial-like cells, while the other is cobblestone-like ceils. Therefore, TSOC1 and TSOC2 appear to consist of ovarian somatic cells. Growth requirement condition was investigated including temperature, concentration of FBS and pH. Significant fluorescent signals were observed after TSOC1 and TSOC2 cells were transfected with pEGFP-N3 vector, indicating its potential utility for genetic manipulation such as gene function studies. It is shown that these cell lines are effective for infection by the turbot reddish body iridovirus and flounder lymphosystis disease virus as evidenced by the appearance of cytopathic effect and virus propagation in the virus-infected cells, and most convincingly, the observation of viral particles by electron microscopy, demonstrating that TSOC1 and TSOC2 are suitable to study interactions between virus and host cells. It is believed that TSOC1 and TSOC2 will be useful tools to study sex-related events and interactions between primordial germ cells and oogonia cells during oogenesis. Therefore, establishment of ovary cell lines from Fugu rubripes seems to be significant for those research areas.
