Prostaglandin E2(PGE2)has been reported to be involved in reproduction as one of the most abundant and widely distributed prostaglandins in invertebrates.Regulating gonad development and gamete production in mollusks ...Prostaglandin E2(PGE2)has been reported to be involved in reproduction as one of the most abundant and widely distributed prostaglandins in invertebrates.Regulating gonad development and gamete production in mollusks can greatly assist artificial breeding,which is believed to promote the commercial benefits of various mollusk species.In the present study,the contents of PGE2 in the gonads of abalone at different developmental stages(maturing stage,ripe stage,and spawning stage)and reproduction states(before and after spawning)were measured.A higher PGE2 level was found in the gonads of abalones at the ripe stage than other stages,and the highest PGE2 level was detected in the ovaries of female H.diversicolor in the UV-stimulated group.Then the effects of PGE2 injection on the spawning efficiency were explored,and the gamete emission rates of female and male abalone were significantly increased after PGE2 injection.Through qRT-PCR and in situ hybridization assay,higher expression levels and wide distribution of phospholipase A2(PLA2),cyclooxygenase 2(COX2),prostaglandin E synthases(PGES2),and prostaglandin E2 receptor 4(EP4)in the gonads of H.diversicolor were observed after spawning.It is speculated that PGE2 may promote the reproductive cycle in both male and female H.diversicolor,although the regulation mechanism of PGE2 in female and male abalone might be different.The results will be useful for the application of PGE2 in the abalone aquaculture practices to increase the reproduction efficiency in future.展开更多
The peritoneum is mainly composed of mesothelial cells,fiber cells,and fibers,with many free cells that come from tissue capillaries.Siniperca chuatsi is a valuable fish commodity and important freshwater aquaculture ...The peritoneum is mainly composed of mesothelial cells,fiber cells,and fibers,with many free cells that come from tissue capillaries.Siniperca chuatsi is a valuable fish commodity and important freshwater aquaculture species in China.With the increase in the scale of its breeding,bacterial and viral disease of the S.chuatsi has become increasingly rampant in recent years.This study analyzed the peritoneal gene expression profile using transcriptome technology for the first time.The results showed that 95,711 unigenes were expressed in the peritoneum with an average length of 920.72 bp.In addition to the housekeeping genes,the genes of extracellular matrix protein 1a(ecm1),Angiopoietin-related protein 4-like(angptl4)and cysteine-rich protein 1(crip1)were highly expressed genes in the peritoneum,and they are key factors in peritoneal development and the maintenance of morphology.Meanwhile,many immune genes were also expressed in the peritoneum,including beta-2-microglobulin(b2m),MHC II invariant chain(cd74),prostate stem cell antigen-like protein(psca),claudin(cldn),heat shock protein 90(hsp90),complement component 4(c4),radical S-adenosyl methionine domain-containing protein 2(rsad2),and cathepsin S(ctss).The RT-PCR results showed that different genes exhibited different expression profiles.The transcriptome data indicated that b2m,cldn,hsp90,c4,and rsad2 were highly expressed in the peritoneum.b2m,cd74,psca,c4,and ctss were highly expressed after embryonic hatching,whereas cldn,hsp90,and rsad2 were expressed at early embryonic stages.Quantitative PCR demonstrated that b2m,cd74,cldn,hsp90,c4,and ctss genes were highly expressed after immunostimulation.This study provides significant data for studying the morphological and functional diversity of fish peritoneum and lays a foundation for research on mandarin fish immunity.展开更多
The erythrocytes or red blood cells(RBCs)and leucocytes or white blood cells(WBCs)are the most abundant cell types in the circulatory system of vertebrates.The RBCs are responsible for gas exchange while WBCs are sole...The erythrocytes or red blood cells(RBCs)and leucocytes or white blood cells(WBCs)are the most abundant cell types in the circulatory system of vertebrates.The RBCs are responsible for gas exchange while WBCs are solely involved with immunity.In this study,we present evidence that fish RBCs despite their respiratory role they also express immune-related genes and responses in the teleost fish,Nile tilapia(Oreochromis niloticus).Fluorescent staining identified the presence of numerous mitochondria and lysosomes in tilapia RBCs.RNA sequencing of tilapia RBCs and WBCs led to the assembly and annotation of 20.876 transcripts of which 1017 and 3628 transcripts were specific to RBCs and WBCs,respectively.Pairwise sequence comparisons identified 3251 transcripts that were significantly differential expressed of which 707(21.75%)were up-regulated and 2544(78.25%)down-regulated in RBCs when compared to WBCs.809 transcripts were assigned to 16 immune system pathways,21 and 163 transcripts were specifically expressed in tilapia RBCs and WBCs respectively,and 23 and 233 transcripts were highly expressed in RBCs and WBCs respectively.Expression of 7 immune genes(interferon regulatory factors(irf)1e5 and 9 and tlr3)in both WBCs and RBCs was validated by RT-PCR analysis.Quantitative real-time PCR revealed that all genes were up-regulated in poly(I:C)-challenged WBCs and irf(1,3,4,and 9)and tlr3 were also significantly up-regulated in poly(I:C)-challenged RBCs.Therefore,fish RBCs expressed immunity genes and responses and they may play a complementary role in vertebrate immunity.展开更多
MicroRNAs(miRNAs)are short noncoding sequences that can regulate the expression of target genes.Erythrocytes,also called red blood cells(RBCs),are the largest proportion of cells in blood.Erythrocytes deliver oxygen i...MicroRNAs(miRNAs)are short noncoding sequences that can regulate the expression of target genes.Erythrocytes,also called red blood cells(RBCs),are the largest proportion of cells in blood.Erythrocytes deliver oxygen in the body of vertebrates and contribute to the body’s immune function.Illumina sequencing technology was used to analyze pooled RNA samples isolated from erythrocytes,in order to characterize their gene expression profile and increase the range of miRNAs identified in Nile tilapia erythrocytes.This study identified conserved(n=309)and novel(n=194)miRNAs.Quantitative reverse-transcription polymerase chain reaction and next-generation sequencing results indicated that miR-451 was the most abundant in RBCs,followed by let-7a-1,miR-2188 and miR-144.Moreover,the tissue expression mode of these highly expressed miRNAs was characterized.Results show that miR-451,miR-2188,and miR-144 were all highly expressed in RBCs but were low abundance in other tissues.miR-1 and miR-10b were also abundantly expressed in muscle,while miR-122 was abundantly expressed in the liver.let-7a-1,miR-10b,and miR-1 were significantly higher in the testis than in the ovary.The tissue distribution of miRNAs implies that these noncoding sequences have a potential role in regulating tissue differentiation or maintaining tissue characteristics.The results of a dual-luciferase reporter assay indicated that miR-144 and miR-10b inhibited the expression of the meningioma expressed antigen 5 gene in vitro.This study contributes to the Nile tilapia miRNA database and reveals the profile of erythrocyte miRNA expression.展开更多
The Lateolabrax maculatus is an important marine fish in China that can adapt to a wide range of water salinities.The gill is the main organ involved in fish osmoregulation and in this study,a cDNA library of the L.ma...The Lateolabrax maculatus is an important marine fish in China that can adapt to a wide range of water salinities.The gill is the main organ involved in fish osmoregulation and in this study,a cDNA library of the L.maculatus gill was constructed to understand gill function in Lateolabrax maculatus and it adaptation to salinity.A total raw data of 5.3G base pairs was obtained,followed removal of low-quality and linker DNA sequences and it yielded a total of 23,038,590 clean reads.After splicing and clustering,31,976 unigenes were obtained.The average transcript length was 1327 bp and N50 was 2057 bp.In comparisons with other tissues analysis of the gill transcriptome revealed a high level of relative expression of apolipoprotein E,CD74,keratin,type I cytoskeleton 13,carbonic anhydrase,betaine-homocysteine S-methyltransferase,and aquaporin 3(AQP3)genes.The L.maculatus aqp3(Lm-aqp3)gene was specific for the gills Lm-aqp3 and this gene has a typical transmembrane structure of six alpha helices and a water channel domain.Phylogenetic analysis indicated that Lm-aqp3 clusters with the homologue genes from Labrus bergylta and Dicentrarchus labrax.Quantitative RT-PCR confirmed that Lm-aqp3 was highly abundant in the gills and low expressed in other tissues(stomach,liver,brain,spleen and testis).Analysis of Lm-aqp3 in freshwater aquaculture fish revealed that gene expression was significantly statistical higher than in seawater(20 ppt)fish,suggesting that Lm-aqp3 may play an important role in water metabolism and osmotic pressure regulation in L.maculatus.展开更多
The Chinese mitten crab(Eriocheir sinensis)possesses an open circulatory system,in which hemolymph moves through interconnected sinuses or spaces surrounding organs called hemocoels.The hemocytes are classified into h...The Chinese mitten crab(Eriocheir sinensis)possesses an open circulatory system,in which hemolymph moves through interconnected sinuses or spaces surrounding organs called hemocoels.The hemocytes are classified into hyalinocytes,semigranulocytes,and granulocytes;and limited transcriptomics research is available.In this study,the transcriptome of the crab E.sinensis hemocytes was characterized.A total of 14,380,229 clean reads representing a total of 4.31 Gb nucleotides dataset were produced.A total of 67,047 contigs were obtained and 12.49%(8375)and 9.74%(6533)of the contigs were matched to data publicly available from the GenBank nr nucleotide and Uniprot databases,respectively.Among these contigs,4344 contigs belong to three categories of Gene Ontology,126 contigs to 21 subcategories of KEGG,and 4962 contigs to 25 categories of the COG database.A total of 508,379 and 345 transcripts of the E.sinensis showed>40%sequence similarity with transcripts expressed in the vertebrate blood cells from tilapia(Oreochromis niloticus),Chinese softshell turtle(Trionyx sinensis)and chicken(Gallus gallus)respectively.A total of 53,077 single nucleotide polymorphisms were identified and 9912 SSRs for microsatellite mining were found.The most frequent repeat motifs detected were dinucleotide repeats,accounting for 37.52%of the total repeats.Our data provides an important gene resource for the exploitation of molecular marker-assisted breeding,immune mechanisms and conservation of germplasm in the crab E.sinensis.展开更多
基金supported by grants from National Natural Science Foundation of China (No.31860728 and 32172961)Special Fund for Ocean and Fisheries of Xiamen (No.21CZY018HJ01).
文摘Prostaglandin E2(PGE2)has been reported to be involved in reproduction as one of the most abundant and widely distributed prostaglandins in invertebrates.Regulating gonad development and gamete production in mollusks can greatly assist artificial breeding,which is believed to promote the commercial benefits of various mollusk species.In the present study,the contents of PGE2 in the gonads of abalone at different developmental stages(maturing stage,ripe stage,and spawning stage)and reproduction states(before and after spawning)were measured.A higher PGE2 level was found in the gonads of abalones at the ripe stage than other stages,and the highest PGE2 level was detected in the ovaries of female H.diversicolor in the UV-stimulated group.Then the effects of PGE2 injection on the spawning efficiency were explored,and the gamete emission rates of female and male abalone were significantly increased after PGE2 injection.Through qRT-PCR and in situ hybridization assay,higher expression levels and wide distribution of phospholipase A2(PLA2),cyclooxygenase 2(COX2),prostaglandin E synthases(PGES2),and prostaglandin E2 receptor 4(EP4)in the gonads of H.diversicolor were observed after spawning.It is speculated that PGE2 may promote the reproductive cycle in both male and female H.diversicolor,although the regulation mechanism of PGE2 in female and male abalone might be different.The results will be useful for the application of PGE2 in the abalone aquaculture practices to increase the reproduction efficiency in future.
基金supported by grants from the China Agriculture Research System (CARS-46)the Key national and special project of blue granary science and technology innovation (2020YFD0900400).
文摘The peritoneum is mainly composed of mesothelial cells,fiber cells,and fibers,with many free cells that come from tissue capillaries.Siniperca chuatsi is a valuable fish commodity and important freshwater aquaculture species in China.With the increase in the scale of its breeding,bacterial and viral disease of the S.chuatsi has become increasingly rampant in recent years.This study analyzed the peritoneal gene expression profile using transcriptome technology for the first time.The results showed that 95,711 unigenes were expressed in the peritoneum with an average length of 920.72 bp.In addition to the housekeeping genes,the genes of extracellular matrix protein 1a(ecm1),Angiopoietin-related protein 4-like(angptl4)and cysteine-rich protein 1(crip1)were highly expressed genes in the peritoneum,and they are key factors in peritoneal development and the maintenance of morphology.Meanwhile,many immune genes were also expressed in the peritoneum,including beta-2-microglobulin(b2m),MHC II invariant chain(cd74),prostate stem cell antigen-like protein(psca),claudin(cldn),heat shock protein 90(hsp90),complement component 4(c4),radical S-adenosyl methionine domain-containing protein 2(rsad2),and cathepsin S(ctss).The RT-PCR results showed that different genes exhibited different expression profiles.The transcriptome data indicated that b2m,cldn,hsp90,c4,and rsad2 were highly expressed in the peritoneum.b2m,cd74,psca,c4,and ctss were highly expressed after embryonic hatching,whereas cldn,hsp90,and rsad2 were expressed at early embryonic stages.Quantitative PCR demonstrated that b2m,cd74,cldn,hsp90,c4,and ctss genes were highly expressed after immunostimulation.This study provides significant data for studying the morphological and functional diversity of fish peritoneum and lays a foundation for research on mandarin fish immunity.
基金This work was supported by China Agriculture Research System(CARS-46)Shanghai Collaborate Innovation Center for Aquatic Animal Genetics and Breeding(ZF1206)by the Open Fund of Key Laboratory of Experimental Marine Biology from the Chinese Academy of Sciences(No.KF2017NO2).
文摘The erythrocytes or red blood cells(RBCs)and leucocytes or white blood cells(WBCs)are the most abundant cell types in the circulatory system of vertebrates.The RBCs are responsible for gas exchange while WBCs are solely involved with immunity.In this study,we present evidence that fish RBCs despite their respiratory role they also express immune-related genes and responses in the teleost fish,Nile tilapia(Oreochromis niloticus).Fluorescent staining identified the presence of numerous mitochondria and lysosomes in tilapia RBCs.RNA sequencing of tilapia RBCs and WBCs led to the assembly and annotation of 20.876 transcripts of which 1017 and 3628 transcripts were specific to RBCs and WBCs,respectively.Pairwise sequence comparisons identified 3251 transcripts that were significantly differential expressed of which 707(21.75%)were up-regulated and 2544(78.25%)down-regulated in RBCs when compared to WBCs.809 transcripts were assigned to 16 immune system pathways,21 and 163 transcripts were specifically expressed in tilapia RBCs and WBCs respectively,and 23 and 233 transcripts were highly expressed in RBCs and WBCs respectively.Expression of 7 immune genes(interferon regulatory factors(irf)1e5 and 9 and tlr3)in both WBCs and RBCs was validated by RT-PCR analysis.Quantitative real-time PCR revealed that all genes were up-regulated in poly(I:C)-challenged WBCs and irf(1,3,4,and 9)and tlr3 were also significantly up-regulated in poly(I:C)-challenged RBCs.Therefore,fish RBCs expressed immunity genes and responses and they may play a complementary role in vertebrate immunity.
基金This work was supported by the China Agriculture Research System(CARS-46)Shanghai Collaborate Innovation Center for Aquatic Animal Genetics and Breeding(ZF1206).
文摘MicroRNAs(miRNAs)are short noncoding sequences that can regulate the expression of target genes.Erythrocytes,also called red blood cells(RBCs),are the largest proportion of cells in blood.Erythrocytes deliver oxygen in the body of vertebrates and contribute to the body’s immune function.Illumina sequencing technology was used to analyze pooled RNA samples isolated from erythrocytes,in order to characterize their gene expression profile and increase the range of miRNAs identified in Nile tilapia erythrocytes.This study identified conserved(n=309)and novel(n=194)miRNAs.Quantitative reverse-transcription polymerase chain reaction and next-generation sequencing results indicated that miR-451 was the most abundant in RBCs,followed by let-7a-1,miR-2188 and miR-144.Moreover,the tissue expression mode of these highly expressed miRNAs was characterized.Results show that miR-451,miR-2188,and miR-144 were all highly expressed in RBCs but were low abundance in other tissues.miR-1 and miR-10b were also abundantly expressed in muscle,while miR-122 was abundantly expressed in the liver.let-7a-1,miR-10b,and miR-1 were significantly higher in the testis than in the ovary.The tissue distribution of miRNAs implies that these noncoding sequences have a potential role in regulating tissue differentiation or maintaining tissue characteristics.The results of a dual-luciferase reporter assay indicated that miR-144 and miR-10b inhibited the expression of the meningioma expressed antigen 5 gene in vitro.This study contributes to the Nile tilapia miRNA database and reveals the profile of erythrocyte miRNA expression.
基金China Agriculture Research System(CARS-47)and Open Fund of Key Laboratory of Experimental Marine Biology,Chinese Academy of Sciences(KF2017NO2).
文摘The Lateolabrax maculatus is an important marine fish in China that can adapt to a wide range of water salinities.The gill is the main organ involved in fish osmoregulation and in this study,a cDNA library of the L.maculatus gill was constructed to understand gill function in Lateolabrax maculatus and it adaptation to salinity.A total raw data of 5.3G base pairs was obtained,followed removal of low-quality and linker DNA sequences and it yielded a total of 23,038,590 clean reads.After splicing and clustering,31,976 unigenes were obtained.The average transcript length was 1327 bp and N50 was 2057 bp.In comparisons with other tissues analysis of the gill transcriptome revealed a high level of relative expression of apolipoprotein E,CD74,keratin,type I cytoskeleton 13,carbonic anhydrase,betaine-homocysteine S-methyltransferase,and aquaporin 3(AQP3)genes.The L.maculatus aqp3(Lm-aqp3)gene was specific for the gills Lm-aqp3 and this gene has a typical transmembrane structure of six alpha helices and a water channel domain.Phylogenetic analysis indicated that Lm-aqp3 clusters with the homologue genes from Labrus bergylta and Dicentrarchus labrax.Quantitative RT-PCR confirmed that Lm-aqp3 was highly abundant in the gills and low expressed in other tissues(stomach,liver,brain,spleen and testis).Analysis of Lm-aqp3 in freshwater aquaculture fish revealed that gene expression was significantly statistical higher than in seawater(20 ppt)fish,suggesting that Lm-aqp3 may play an important role in water metabolism and osmotic pressure regulation in L.maculatus.
基金This research was supported by General Project from the Natural Science Foundation of China(No.31572630)The capacity promoting project of Shanghai Engineering and Technology Center from Shanghai Municipal Science and Technology Commission(No.16DZ2281200)Shanghai Universities Top Disciplines Project of Fisheries(No.2015-0908)from Shanghai Education Commission。
文摘The Chinese mitten crab(Eriocheir sinensis)possesses an open circulatory system,in which hemolymph moves through interconnected sinuses or spaces surrounding organs called hemocoels.The hemocytes are classified into hyalinocytes,semigranulocytes,and granulocytes;and limited transcriptomics research is available.In this study,the transcriptome of the crab E.sinensis hemocytes was characterized.A total of 14,380,229 clean reads representing a total of 4.31 Gb nucleotides dataset were produced.A total of 67,047 contigs were obtained and 12.49%(8375)and 9.74%(6533)of the contigs were matched to data publicly available from the GenBank nr nucleotide and Uniprot databases,respectively.Among these contigs,4344 contigs belong to three categories of Gene Ontology,126 contigs to 21 subcategories of KEGG,and 4962 contigs to 25 categories of the COG database.A total of 508,379 and 345 transcripts of the E.sinensis showed>40%sequence similarity with transcripts expressed in the vertebrate blood cells from tilapia(Oreochromis niloticus),Chinese softshell turtle(Trionyx sinensis)and chicken(Gallus gallus)respectively.A total of 53,077 single nucleotide polymorphisms were identified and 9912 SSRs for microsatellite mining were found.The most frequent repeat motifs detected were dinucleotide repeats,accounting for 37.52%of the total repeats.Our data provides an important gene resource for the exploitation of molecular marker-assisted breeding,immune mechanisms and conservation of germplasm in the crab E.sinensis.
