Perfluorooctanoic acid(PFOA)is a widespread synthetic persistent organic pollutant that may enrich along the food chain and affect the growth,development,reproduction,and lipid metabolism of aquatic organisms,particul...Perfluorooctanoic acid(PFOA)is a widespread synthetic persistent organic pollutant that may enrich along the food chain and affect the growth,development,reproduction,and lipid metabolism of aquatic organisms,particularly the benthic organisms.How-ever,the toxic effects of PFOA on the half-smooth tongue sole Cynoglossus semilaevis,a commercial benthic fish in China,have rarely been reported.Because juvenile fish are sensitive to environmental pollutants,in the present study,histological assessment and tran-scriptome sequencing were performed to determine the short-term impact of PFOA on juvenile half-smooth tongue soles.Histologi-cal analysis showed that PFOA exposure caused hepatocyte rupture,intestinal villi breakage,increased goblet cell count,and brain ab-normal.Transcriptome results showed that some interesting signaling pathways,such as glycolysis/gluconeogenesis,PPAR signaling pathway and GABAergic synapse signaling pathway,were enriched after PFOA exposure.In addition,some metabolic,immune and neural genes were differentially expressed,which including ependymin,hbb1-like and gad 1,and they were up-regulated after 14 days of exposure.Transcriptome results also indicated that half-smooth tongue sole might improve energy metabolism in response to PFOA toxicity after 7 days of exposure.These findings provide a basis for studying the ecological effects of PFOA on marine benthic fishes.展开更多
A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 ...A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 passages for over 200 d in DMEM medium supplemented with 15% FBS （fetal bovine serum）, 2.5 ng/cm^3 bFGF （basic fibroblast growth factor）, 1 ng/cm^3 LIF （leukemia inhibitory factor） and 50 mmol/dm^3 2-ME （2-mecaptoethanol）. The cells grew well in the temperature range of 24-30 ℃ and the optimal growth temperature was 24 ℃. FBS and bFGF concentrations are the two key components for CSEC cells proliferation. Chromosome analysis reveals that CSEC cells have a normal diploid karyotype with 2n=42t. The significant fluorescent signals were observed in CSEC after transfection with the GFP reporter gene, suggesting that the CSEC cell line can be used as a useful tool for transgenic and genetic manipulation studies. CSEC cells showed the cytopathic effect （CPE） after infection with lymphosystis disease virus （LCDV） in 2 d. Moreover, the LCDV particles can be observed in the cytoplasm of virus-infected cells by electron microscopy. It suggests that CSEC could be potentially used for the study of aquatic virus.展开更多
A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subculturedmore than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ...A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subculturedmore than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1basic fi-broblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.TheSHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃and 30℃with the optimum growth at 24℃and with a reduced growth between 12℃and 20℃.The growth rate of the cells increased as the proportion ofFBS increased from 10%to 20%at 28℃with optimum growth at the concentration of 20%.The doubling time of the cells was de-termined to be 44.8 h.Chromosome analysis revealed that 52%of the SHK cells maintained a normal diploid chromosome number(2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFPgene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis diseasevirus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCRand electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.展开更多
基金supported by the National Key R&D Program(No.2018YFD0900301-03)the MNR Key Laboratory of Marine Eco-Environmental Science and Technology,China(No.MEEST-2021-04).
文摘Perfluorooctanoic acid(PFOA)is a widespread synthetic persistent organic pollutant that may enrich along the food chain and affect the growth,development,reproduction,and lipid metabolism of aquatic organisms,particularly the benthic organisms.How-ever,the toxic effects of PFOA on the half-smooth tongue sole Cynoglossus semilaevis,a commercial benthic fish in China,have rarely been reported.Because juvenile fish are sensitive to environmental pollutants,in the present study,histological assessment and tran-scriptome sequencing were performed to determine the short-term impact of PFOA on juvenile half-smooth tongue soles.Histologi-cal analysis showed that PFOA exposure caused hepatocyte rupture,intestinal villi breakage,increased goblet cell count,and brain ab-normal.Transcriptome results showed that some interesting signaling pathways,such as glycolysis/gluconeogenesis,PPAR signaling pathway and GABAergic synapse signaling pathway,were enriched after PFOA exposure.In addition,some metabolic,immune and neural genes were differentially expressed,which including ependymin,hbb1-like and gad 1,and they were up-regulated after 14 days of exposure.Transcriptome results also indicated that half-smooth tongue sole might improve energy metabolism in response to PFOA toxicity after 7 days of exposure.These findings provide a basis for studying the ecological effects of PFOA on marine benthic fishes.
基金The State "863" High-Technology Research and Development Project of China under contract Nos 2006AA09Z406 and 2006AA10A401the National Natural Science Foundation of China under contract No. 40376047the Taishan Scholar Project of Shandong Province and MOA for returned scientists from abroad
文摘A new cell line, CSEC, has been successfully established from embryos at gastrula stage of a cultured marine fish, half smooth tongue sole （Cynoglossus sernilaevis）. CSEC cells grow actively and stably more than 50 passages for over 200 d in DMEM medium supplemented with 15% FBS （fetal bovine serum）, 2.5 ng/cm^3 bFGF （basic fibroblast growth factor）, 1 ng/cm^3 LIF （leukemia inhibitory factor） and 50 mmol/dm^3 2-ME （2-mecaptoethanol）. The cells grew well in the temperature range of 24-30 ℃ and the optimal growth temperature was 24 ℃. FBS and bFGF concentrations are the two key components for CSEC cells proliferation. Chromosome analysis reveals that CSEC cells have a normal diploid karyotype with 2n=42t. The significant fluorescent signals were observed in CSEC after transfection with the GFP reporter gene, suggesting that the CSEC cell line can be used as a useful tool for transgenic and genetic manipulation studies. CSEC cells showed the cytopathic effect （CPE） after infection with lymphosystis disease virus （LCDV） in 2 d. Moreover, the LCDV particles can be observed in the cytoplasm of virus-infected cells by electron microscopy. It suggests that CSEC could be potentially used for the study of aquatic virus.
基金supported by grants from State 863High-Technology Rand Project of China(2006AA09Z406,2006AA10A401)Taishan Scholar Project of Shan-dong Province
文摘A cell line,SHK,was derived from the kidney of spotted halibut Verasper variegates.The cell line was subculturedmore than 40 passages in minimum essential medium(MEM)supplemented with fetal bovine serum(FBS)and 10 ng ml-1basic fi-broblast growth factor(bFGF).Cell morphology from primary culture and subculture was observed continuously by microscopy.TheSHK cell line consisted predominantly of fibroblast-like cells.The cell line was able to grow between 20℃and 30℃with the optimum growth at 24℃and with a reduced growth between 12℃and 20℃.The growth rate of the cells increased as the proportion ofFBS increased from 10%to 20%at 28℃with optimum growth at the concentration of 20%.The doubling time of the cells was de-termined to be 44.8 h.Chromosome analysis revealed that 52%of the SHK cells maintained a normal diploid chromosome number(2n=46).The cells were successfully transfected with green fluorescent protein(GFP)reporter plasmids and the expression of GFPgene in the cells indicated the possible utility of the cells in gene expression studies.The cells were infected by lymphosystis diseasevirus(LCDV)and found to be susceptible to the virus in cytopathic effect(CPE)observation.The infection was confirmed by PCRand electron microscopy experiments,which proved the existence of the viral particles in the cytoplasm of the virus-infected cells.
