By inserting the CMV with HSV TK promoter into the plasmid pBlue-cGH which contained cGH gene, a chimeric expression vector was constructed for transgenic fish. After the vector was transferred into fertilized eggs of...By inserting the CMV with HSV TK promoter into the plasmid pBlue-cGH which contained cGH gene, a chimeric expression vector was constructed for transgenic fish. After the vector was transferred into fertilized eggs of crucian carp by means of microinjection, the transgenic crucian carp was obtained. The result of PCR detection showed that the growth hormone gene of the exogenous carp was integrated into the genome of crucian carp, and the transgenic crucian carp displayed enhancement of its growth.展开更多
Salt tolerance and freeze preservation of crucian are always important concerns of people. There are rare reports on effect of different salinity on fresh- ness and quality. However, water salinity will directly affec...Salt tolerance and freeze preservation of crucian are always important concerns of people. There are rare reports on effect of different salinity on fresh- ness and quality. However, water salinity will directly affect osmotic pressure in fish, thereby affecting meat quality. In this study, the tolerance of crucian to different salinities （1‰, 2‰, 5‰, 8‰, 10‰ and 15‰） was investigated. In addition, the ef- fect of different salinity on freezing freshness and quality of crucian was studied. The results showed that the tolerance threshold of crucian to salinity was about 8‰, and the freezing quality of crucian in water with salinity of ‰ was significantly better than that in fresh water.展开更多
In order to provide data for joints control of our recently designed crucian hke biomlmetlc robot fish, an A-ray photograph technology was adopted to determine the number and length of vertebral joints. A frame-by-fra...In order to provide data for joints control of our recently designed crucian hke biomlmetlc robot fish, an A-ray photograph technology was adopted to determine the number and length of vertebral joints. A frame-by-frame analysis of high-speed videotapes was conducted to quantify the kinematics of crucian at four speeds （12.651 cm·s^-1, 18.201 cm·s^-1, 21.901 cm·s^-1, 24.368cm·s^-1） during cruising. In addition to a brief introduction to experimental conditions and methods, we analyzed the influence of individual diversity on the absolute length as well as the non-dimensional length of vertebral joints. We also presented the maximal angular velocity and acceleration of vertebral joints under four swimming speeds, and provided the change of relative rotation angle, angular difference, angular velocity and angular acceleration of the rear vertebral joints with time at a certain swimming speed of 12.651 cm·s^-1. At last, we presented the maximal lateral displacement of each mark at that speed. The study found that the influence of individual diversity on the non-dimensional length of vertebral joints is not significant; the maximal angular velocity and acceleration of vertebral joints increase with swimming speed; angular difference, angular velocity and angular acceleration exhibit two maximal values over one period at a certain swimming speed.展开更多
The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimicti...The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimictic colored crucian carp. MPF activity was measured by using histone H1 as phosphorylation substrate. There were two similar oscillatory MPF kinase activity changes during oocyte maturation in two kinds of fishes with different reproductive modes, but there existed some subtle difference between them. The subtle difference was that the first peak of MPF kinase activity was kept to a longerlasting time in the gynogenetic silver crucian carp than in the amphimictic colored crucian carp. It was suggested that the difference may be related to the spindle behavior changes, such as tripolar spindle formation and spindle rearrangement in the gynogenetic crucian carp.展开更多
The kinematics of turning maneuvers of startled Crucian Carp (Carassius auratus) are presented. All escape responses observed are C-type fast-starts. The position of the center of mass and the moment of inertia of the...The kinematics of turning maneuvers of startled Crucian Carp (Carassius auratus) are presented. All escape responses observed are C-type fast-starts. The position of the center of mass and the moment of inertia of the fish are calculated. The results show that the position of the center of mass is always at 35% of the length of the fish from the head and the position of the center of mass and moment of inertia can be considered unchanged during C-start of Crucian Carp. Hydrodynamic analysis of the C-start is given based on the kinematics data from our experiments. The C-start consists of three stages. In stage 1, the tail fin of fish rapidly flaps in one direction, and a large moment acts on the fish′s body, which rotates around the center of mass with an angular acceleration. In stage 2, the tail fin flaps more slowly in the opposite direction at slower speed, the fish′s body rotates around the center of mass with angular deceleration and the center of mass of the fish moves along an arc. In stage 3, the moment approximately equals zero, the fish′s body stops rotating and the center of mass the moves along a straight line.展开更多
The essay studies the influence of the different intensities of fishing stress on body weight （7.30 ± 1.48） g of red crucian carp juvenile body length, weight, fatness, feeding rate, specific growth rate, food ...The essay studies the influence of the different intensities of fishing stress on body weight （7.30 ± 1.48） g of red crucian carp juvenile body length, weight, fatness, feeding rate, specific growth rate, food conversion rate, intestinal protease, amylase and immunoglobulin. The result showed that body length, weight, fatness, feeding rate, specific growth rate of red crucian carp juvenile were significantly inhibited from fishing stress compared with the control group, and the magnitude of the inhibitory effect increased with the time going and increased. Fishing stress could increase the food conversion rate of red crucian carp juvenile, and the different intensity stress had no significant different influence. The intestinal protease activity decreased in the beginning, then increased, and was consistent with the level of the control group ultimately. High-stress group accelerated the decline of intestinal amylase activity of the juvenile. The stress increased the number of immunoglobulin of juvenile red crucian carp significantly, but the increase disappeard with the stress time extended.展开更多
Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenes...Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels,little is currently known regarding the regulation of sperm flagella assembly by microRNAs(miRNAs).The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied.Results:In this study,we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp.As one of the testis-specific candidate markers,Tekt1 was predominantly expressed in the testis.Quantitative real-time PCR(qRT-PCR)analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1.Through bioinformatics analysis,we identified a putative miR-199-5p binding site in the Tekt1 mRNA.We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay.Finally,we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp.Moreover,miR-199-5p gainof-function could lead to spermatids apoptosis and abnormal spermatozoa structure,which is similar to that of allotriploid crucian carp.Conclusions:Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1,thereby causing sperm abnormalities of male allotriploid crucian carp.展开更多
[Objective] To investigate the effects of amino acid powder, fermented rapeseed meal and fermented cottonseed meal on the growth performance, amino acid absorption, body composition and liver function of allogynogenti...[Objective] To investigate the effects of amino acid powder, fermented rapeseed meal and fermented cottonseed meal on the growth performance, amino acid absorption, body composition and liver function of allogynogentic crucian carp. [ Method] The allogynogentic crucian carps with the average initial weight of 17.3 g were divided into control group and six experimental groups randomly. The allogynogentic crucian carps in the control group and experimental groups were fed a basal diet and the basal diets supplemented by amino acid powder, fermented rapeseed meal and fermented cottonseed meal, respectively. The growth rate and specific growth rate of allogynogentic crucian carps, the content of amino acids in serum and feeds, the composition indicators of body, muscle and liver, as well as the activities of glutamic-pyruvic transaminase （GPT） and glutamic-oxaloacetic transaminase （GOT） in liver and serum were determined, respectively. [ Result] The growth rate and specific growth rate （SGR） of allogynogentic crucian carps in the 3% amino acid powder group and the 6% fermented cottonseed meal group were increased greatly （ P 〈 0.05）, and no significant difference was found between experimental group and control group. The serum lysine content of the 3% amino acid powder group and the 6% fermented cottonseed meal group was higher than that of the control group. In the experimental groups, the absorption of four essential amino acids was up to a peak at 4 h after feeding, which was consistent with the control group. The body composition and liver function of allogynogentic crucian carps had no significant difference between the experimental groups and the control group. [ Conclusion] The 3% amino acid powder, 6% fermented cottonseed meal and 6% fermented rapeseed meal can substitute fish meal, cottonseed meal and rapeseed meal to improve the clrowth performance, amino acid absorption of the allogynogentic crucian carps.展开更多
The interest in the analysis of phospholipids(PLs), especially phosphatidylcholine(PC), has been increasing due to the importance of them in biochemistry as well as in industry. A method was reported based on the offl...The interest in the analysis of phospholipids(PLs), especially phosphatidylcholine(PC), has been increasing due to the importance of them in biochemistry as well as in industry. A method was reported based on the offline combination of MALDI-TOF MS and normal-phase HPLC for analyzing PLs extracted from crucian carp. Total PLs of crucian carp were extracted and then separated by HPLC before the collected subfractions were analyzed by MALDI-TOF MS. The mass spectra obtained show peaks of H+, Na+ and K+ adducts of PC molecules. It is shown that the prior separation of PLs by HPLC is highly necessary to remove the signal suppressing and to avoid the possible overlapping. With this method, 9 possible PC molecules in crucian carp and the corresponding fatty acid compositions were given from the well-resolved mass spectra.展开更多
Inorganic lead （Pb） is one of the most common environmental pollutants. Much evidence indicates that Pb exposure could directly affect fish growth and development. In this study, we investigated the cytotoxic effect...Inorganic lead （Pb） is one of the most common environmental pollutants. Much evidence indicates that Pb exposure could directly affect fish growth and development. In this study, we investigated the cytotoxic effects of Pb on cytoskeletal protein stability at both protein and mRNA level in crucian carp Carassius auratus. Pb（NO3）2 treatment in concentration of 100 μmol/L resulted in decreased expression of both α- and β-tubulin but γ-tubulin as assayed with SDS-PAGE, Western Blot, and ELISA. In vivo and in vitro analyses on protein expression of tubulins are consistent. The effect of Pb on mRNA expression varied among different tissues. Our results suggest that cytotoxicity of Pb at protein translation level is stronger than at mRNA expression level.展开更多
Marinate the clams in brine for 3 hours, until silt isdiffused, and keep them fresh. Draw the crucian carp,and make 3 or 4 diagonal cuts on the surface of bothsides.
A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on scre...A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.展开更多
Crucian carp(Carassius auratus gibelio),an extensively cultivated freshwater fish,was one of the model species for the study of fish immunology.Polyclonal antibodies were advantageous molecular tools for studying tele...Crucian carp(Carassius auratus gibelio),an extensively cultivated freshwater fish,was one of the model species for the study of fish immunology.Polyclonal antibodies were advantageous molecular tools for studying teleost immune system.Specifically,polyclonal antibodies reacting with immunoglobulins(Ig)were used successfully in studies of the teleost fishes.In the present study,we produced polyclonal antibody against CH2 domains of crucian carp IgM,and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 infection.The recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic cells.The specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp serum.Quantitative RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 infection.The expression of IgM in the kidney increased and reached a maximum at 6 h post-infection(hpi),while dropped to a low level at 5 days post-infection(dpi).In conclusion,the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2,indicating that IgM played a potential role in systemic immunity against viral infection.Polyclonal antibody against crucian carp IgM had certain clinical relevance,which might provide insight into the early stage of virus infection and prevention of the disease.展开更多
Tender and delicious,crucian carp is one of the favorites of Wuhan locals who love dining on fish.However,it is very annoying to pick out tiny fishbones while eating.Recently,following more than a year of research,a t...Tender and delicious,crucian carp is one of the favorites of Wuhan locals who love dining on fish.However,it is very annoying to pick out tiny fishbones while eating.Recently,following more than a year of research,a team of academicians led by Gui Jianfang of the Institute of Hydrobiology at the Chinese Academy of Sciences has now released its latest findings related to this issue-the boneless crucian fish has been created.展开更多
In this study, we conducted a cross of white crucian carp （♀）xred crucian carp （♂） （WR）, and characterized the morphology, reproduction and genetics of the progeny. Different from parents, WR with the gray col...In this study, we conducted a cross of white crucian carp （♀）xred crucian carp （♂） （WR）, and characterized the morphology, reproduction and genetics of the progeny. Different from parents, WR with the gray color showed the hybrid morphological traits of both parents. WR possessed normal gonads producing mature eggs or sperm, and exhibited high fertilization rate （90.2%） and high hatchery rate （80.5%）, which contributed to produce and enlarge the population. WR with the same DNA content as parents was a diploid fish with 100 chromosomes （2n=100）. Amplified ITS of 45S rDNA, in WR the sequences consisting of 884 bp bases of the entire ITS-1 region, 5.8 S region, and entire ITS-2 region. The sequences showed high similarity between WR and its parents and leaned towards male inheritance. In WR, NTS of 5S rDNA consisted of three length types with total 654 bp bases. From sequence analysis of NTS, WR shared 94.2% and 95.1% similarities with their female and male parent, respectively. Sequence analysis of ITS and NTS revealed that there existed recombination and variation in the hybrid progeny, which was the genetic base for adaptation and speciation. In conclusion, we obtained WR from hybridization and it exhibited hybrid traits in morphology and variation in genetic composition showing essential difference with its parents. The obtainment of WR has important significance in fish genetic breeding.展开更多
文摘By inserting the CMV with HSV TK promoter into the plasmid pBlue-cGH which contained cGH gene, a chimeric expression vector was constructed for transgenic fish. After the vector was transferred into fertilized eggs of crucian carp by means of microinjection, the transgenic crucian carp was obtained. The result of PCR detection showed that the growth hormone gene of the exogenous carp was integrated into the genome of crucian carp, and the transgenic crucian carp displayed enhancement of its growth.
基金Supported by Incentive Program Subsidized by Department of Science and Technology of Hebei Province(15926620H)Key Technology Research and Development Program of Science and Technology Bureau of Qinhuangdao City(201401A067201402B043)~~
文摘Salt tolerance and freeze preservation of crucian are always important concerns of people. There are rare reports on effect of different salinity on fresh- ness and quality. However, water salinity will directly affect osmotic pressure in fish, thereby affecting meat quality. In this study, the tolerance of crucian to different salinities （1‰, 2‰, 5‰, 8‰, 10‰ and 15‰） was investigated. In addition, the ef- fect of different salinity on freezing freshness and quality of crucian was studied. The results showed that the tolerance threshold of crucian to salinity was about 8‰, and the freezing quality of crucian in water with salinity of ‰ was significantly better than that in fresh water.
文摘In order to provide data for joints control of our recently designed crucian hke biomlmetlc robot fish, an A-ray photograph technology was adopted to determine the number and length of vertebral joints. A frame-by-frame analysis of high-speed videotapes was conducted to quantify the kinematics of crucian at four speeds （12.651 cm·s^-1, 18.201 cm·s^-1, 21.901 cm·s^-1, 24.368cm·s^-1） during cruising. In addition to a brief introduction to experimental conditions and methods, we analyzed the influence of individual diversity on the absolute length as well as the non-dimensional length of vertebral joints. We also presented the maximal angular velocity and acceleration of vertebral joints under four swimming speeds, and provided the change of relative rotation angle, angular difference, angular velocity and angular acceleration of the rear vertebral joints with time at a certain swimming speed of 12.651 cm·s^-1. At last, we presented the maximal lateral displacement of each mark at that speed. The study found that the influence of individual diversity on the non-dimensional length of vertebral joints is not significant; the maximal angular velocity and acceleration of vertebral joints increase with swimming speed; angular difference, angular velocity and angular acceleration exhibit two maximal values over one period at a certain swimming speed.
文摘The spindle behavior and MPF activity changes in the progression of oocyte maturation were investigated and compared with cytological observation and kinase assay between gynogenetic silver crucian carp and amphimictic colored crucian carp. MPF activity was measured by using histone H1 as phosphorylation substrate. There were two similar oscillatory MPF kinase activity changes during oocyte maturation in two kinds of fishes with different reproductive modes, but there existed some subtle difference between them. The subtle difference was that the first peak of MPF kinase activity was kept to a longerlasting time in the gynogenetic silver crucian carp than in the amphimictic colored crucian carp. It was suggested that the difference may be related to the spindle behavior changes, such as tripolar spindle formation and spindle rearrangement in the gynogenetic crucian carp.
基金This work was supported by the National Natural Science Foundation(No.10332040,10072063)the Innovation Project of the Chinese Academy of Sciences(No.KJCX—SW-L04).
文摘The kinematics of turning maneuvers of startled Crucian Carp (Carassius auratus) are presented. All escape responses observed are C-type fast-starts. The position of the center of mass and the moment of inertia of the fish are calculated. The results show that the position of the center of mass is always at 35% of the length of the fish from the head and the position of the center of mass and moment of inertia can be considered unchanged during C-start of Crucian Carp. Hydrodynamic analysis of the C-start is given based on the kinematics data from our experiments. The C-start consists of three stages. In stage 1, the tail fin of fish rapidly flaps in one direction, and a large moment acts on the fish′s body, which rotates around the center of mass with an angular acceleration. In stage 2, the tail fin flaps more slowly in the opposite direction at slower speed, the fish′s body rotates around the center of mass with angular deceleration and the center of mass of the fish moves along an arc. In stage 3, the moment approximately equals zero, the fish′s body stops rotating and the center of mass the moves along a straight line.
文摘The essay studies the influence of the different intensities of fishing stress on body weight （7.30 ± 1.48） g of red crucian carp juvenile body length, weight, fatness, feeding rate, specific growth rate, food conversion rate, intestinal protease, amylase and immunoglobulin. The result showed that body length, weight, fatness, feeding rate, specific growth rate of red crucian carp juvenile were significantly inhibited from fishing stress compared with the control group, and the magnitude of the inhibitory effect increased with the time going and increased. Fishing stress could increase the food conversion rate of red crucian carp juvenile, and the different intensity stress had no significant different influence. The intestinal protease activity decreased in the beginning, then increased, and was consistent with the level of the control group ultimately. High-stress group accelerated the decline of intestinal amylase activity of the juvenile. The stress increased the number of immunoglobulin of juvenile red crucian carp significantly, but the increase disappeard with the stress time extended.
基金supported by the National Natural Science Foundation of China(Grant No.31872551)Laboratory of Lingnan Modern Agriculture Project(Grant No.NT2021008)+4 种基金the Natural Science Foundation of Hunan Province for Distinguished Young Scholars(Grant No.2020JJ2022)Hunan Provincial Innovation Foundation for Postgraduate(Grant No.CX20200480)the earmarked fund for China Agriculture Research System(Grant No.CARS-45)the Key Research and Development Program of Hunan Province(Grant No.2020NK2016)111 Project(D20007).
文摘Background:Sperm abnormalities are one of the primary factors leading to male sterility,but their pathogenesis is still unclear.Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels,little is currently known regarding the regulation of sperm flagella assembly by microRNAs(miRNAs).The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied.Results:In this study,we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp.As one of the testis-specific candidate markers,Tekt1 was predominantly expressed in the testis.Quantitative real-time PCR(qRT-PCR)analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1.Through bioinformatics analysis,we identified a putative miR-199-5p binding site in the Tekt1 mRNA.We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay.Finally,we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp.Moreover,miR-199-5p gainof-function could lead to spermatids apoptosis and abnormal spermatozoa structure,which is similar to that of allotriploid crucian carp.Conclusions:Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1,thereby causing sperm abnormalities of male allotriploid crucian carp.
基金supported by the Applied Basic Research Projects of Suzhou City (YJG0912)
文摘[Objective] To investigate the effects of amino acid powder, fermented rapeseed meal and fermented cottonseed meal on the growth performance, amino acid absorption, body composition and liver function of allogynogentic crucian carp. [ Method] The allogynogentic crucian carps with the average initial weight of 17.3 g were divided into control group and six experimental groups randomly. The allogynogentic crucian carps in the control group and experimental groups were fed a basal diet and the basal diets supplemented by amino acid powder, fermented rapeseed meal and fermented cottonseed meal, respectively. The growth rate and specific growth rate of allogynogentic crucian carps, the content of amino acids in serum and feeds, the composition indicators of body, muscle and liver, as well as the activities of glutamic-pyruvic transaminase （GPT） and glutamic-oxaloacetic transaminase （GOT） in liver and serum were determined, respectively. [ Result] The growth rate and specific growth rate （SGR） of allogynogentic crucian carps in the 3% amino acid powder group and the 6% fermented cottonseed meal group were increased greatly （ P 〈 0.05）, and no significant difference was found between experimental group and control group. The serum lysine content of the 3% amino acid powder group and the 6% fermented cottonseed meal group was higher than that of the control group. In the experimental groups, the absorption of four essential amino acids was up to a peak at 4 h after feeding, which was consistent with the control group. The body composition and liver function of allogynogentic crucian carps had no significant difference between the experimental groups and the control group. [ Conclusion] The 3% amino acid powder, 6% fermented cottonseed meal and 6% fermented rapeseed meal can substitute fish meal, cottonseed meal and rapeseed meal to improve the clrowth performance, amino acid absorption of the allogynogentic crucian carps.
基金Supported by the Knowledge Innovation Program of the Chinese Academy of Sciences(No.KSCX2-YW-N-035)Projects of Shenyang City Science and Technology Burea, China(Nos.F10-224-4-00, 1091239-6-00)
文摘The interest in the analysis of phospholipids(PLs), especially phosphatidylcholine(PC), has been increasing due to the importance of them in biochemistry as well as in industry. A method was reported based on the offline combination of MALDI-TOF MS and normal-phase HPLC for analyzing PLs extracted from crucian carp. Total PLs of crucian carp were extracted and then separated by HPLC before the collected subfractions were analyzed by MALDI-TOF MS. The mass spectra obtained show peaks of H+, Na+ and K+ adducts of PC molecules. It is shown that the prior separation of PLs by HPLC is highly necessary to remove the signal suppressing and to avoid the possible overlapping. With this method, 9 possible PC molecules in crucian carp and the corresponding fatty acid compositions were given from the well-resolved mass spectra.
基金Supported by National Natural Science Foundation of China (No.30571414,No 30640015)Changsha Municipal Project of Science and Technology (No.K051127-72).
文摘Inorganic lead （Pb） is one of the most common environmental pollutants. Much evidence indicates that Pb exposure could directly affect fish growth and development. In this study, we investigated the cytotoxic effects of Pb on cytoskeletal protein stability at both protein and mRNA level in crucian carp Carassius auratus. Pb（NO3）2 treatment in concentration of 100 μmol/L resulted in decreased expression of both α- and β-tubulin but γ-tubulin as assayed with SDS-PAGE, Western Blot, and ELISA. In vivo and in vitro analyses on protein expression of tubulins are consistent. The effect of Pb on mRNA expression varied among different tissues. Our results suggest that cytotoxicity of Pb at protein translation level is stronger than at mRNA expression level.
文摘Marinate the clams in brine for 3 hours, until silt isdiffused, and keep them fresh. Draw the crucian carp,and make 3 or 4 diagonal cuts on the surface of bothsides.
基金This work was supported by the National Nat-ural Science Foundation of China(Grant No.30130240),the Chinese Academy of Sciences(GrantNo.KSCX2-SW-303).
文摘A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.
基金supported by the General Fund Project of China Shanghai Natural Science Foundation(Program No.22ZR1427200).
文摘Crucian carp(Carassius auratus gibelio),an extensively cultivated freshwater fish,was one of the model species for the study of fish immunology.Polyclonal antibodies were advantageous molecular tools for studying teleost immune system.Specifically,polyclonal antibodies reacting with immunoglobulins(Ig)were used successfully in studies of the teleost fishes.In the present study,we produced polyclonal antibody against CH2 domains of crucian carp IgM,and measured the in vivo dynamics of IgM mRNA caused by CyHV-2 infection.The recombinant protein IgM with relative molecular weight about 53 KD was correctly expressed in prokaryotic cells.The specificity of the polyclonal antibody was evaluated by Western blotting and results revealed that the antibody not only specifically recognized crucian carp serum but also cross-reacted with grass carp serum.Quantitative RT-PCR analysis demonstrated the expression of IgM mRNA changed significantly after CyHV-2 infection.The expression of IgM in the kidney increased and reached a maximum at 6 h post-infection(hpi),while dropped to a low level at 5 days post-infection(dpi).In conclusion,the expression of IgM was significantly upregulated in the kidney of crucian carp infected with CyHV-2,indicating that IgM played a potential role in systemic immunity against viral infection.Polyclonal antibody against crucian carp IgM had certain clinical relevance,which might provide insight into the early stage of virus infection and prevention of the disease.
文摘Tender and delicious,crucian carp is one of the favorites of Wuhan locals who love dining on fish.However,it is very annoying to pick out tiny fishbones while eating.Recently,following more than a year of research,a team of academicians led by Gui Jianfang of the Institute of Hydrobiology at the Chinese Academy of Sciences has now released its latest findings related to this issue-the boneless crucian fish has been created.
基金supported by Major International Cooperation Projects of the National Natural Science Foundation of China(31210103918)the National Natural Science Foundation of China(31201980,31430088 and 31272651)+5 种基金the National High Technology Research and Development Program of China(2011AA100403)the Natural Science Foundation of Hunan Province,China(14JJ3072)Science-Technology Foundation of Hunan Province,China(2014FJ3084)Research Foundation of Education Bureau of Hunan Province,China(13C523)the Cooperative Innovation Center of Engineering and New Products for Developmental Biology of Hunan Province(20134486)the Construct Program of the Key Discipline in Hunan Province,and Construct Program of the National Key Discipline
文摘In this study, we conducted a cross of white crucian carp （♀）xred crucian carp （♂） （WR）, and characterized the morphology, reproduction and genetics of the progeny. Different from parents, WR with the gray color showed the hybrid morphological traits of both parents. WR possessed normal gonads producing mature eggs or sperm, and exhibited high fertilization rate （90.2%） and high hatchery rate （80.5%）, which contributed to produce and enlarge the population. WR with the same DNA content as parents was a diploid fish with 100 chromosomes （2n=100）. Amplified ITS of 45S rDNA, in WR the sequences consisting of 884 bp bases of the entire ITS-1 region, 5.8 S region, and entire ITS-2 region. The sequences showed high similarity between WR and its parents and leaned towards male inheritance. In WR, NTS of 5S rDNA consisted of three length types with total 654 bp bases. From sequence analysis of NTS, WR shared 94.2% and 95.1% similarities with their female and male parent, respectively. Sequence analysis of ITS and NTS revealed that there existed recombination and variation in the hybrid progeny, which was the genetic base for adaptation and speciation. In conclusion, we obtained WR from hybridization and it exhibited hybrid traits in morphology and variation in genetic composition showing essential difference with its parents. The obtainment of WR has important significance in fish genetic breeding.
