Neuropeptides are widely distributed in vertebrates and invertebrates,regulating a variety of physiological activities in the organisms,such as metabolism,feeding and reproduction.In this study,to explore the function...Neuropeptides are widely distributed in vertebrates and invertebrates,regulating a variety of physiological activities in the organisms,such as metabolism,feeding and reproduction.In this study,to explore the function of neuropeptide LFRFamide in Sepia pharaonis,the full-length cDNA of LFRFamide-like gene(named SpLFRFL,MG869822.1)was identified with rapid amplification of cDNA ends(RACE)method.The sequence of SpLFRFL was 860 bp in length and encoded 188 amino acids containing 4 different mature peptides:1 copy of PHTPFRFamide,NSLFRFamide,TIFRFamide,and 3 copies of GNLFRFamide.Multiple alignment and phylogenetic analysis results showed that SpLFRFL shared high identity with LFRFamides of Sepia officinalis and Sepiella japonica and had the closest relationship with them.Through quantitative Real-time PCR(qRT-PCR),it was found that the SpLFRFL gene was highly expressed in the optic lobe and brain at three different stages during gonad development in both genders.Moreover,the four mature peptides at a concentration of 0.01μmol L^(−1) could inhibit the protein synthesis in the Chinese hamster ovary cell strain-K1(CHOK1)induced by SpGnRH.These data suggest that SpLFRFL might be involved in the development and reproduction of S.pharaonis.The results can contribute to future studies on neuropeptide evolution and function and benefit the cuttlefish farming.展开更多
With the rapid development of oil,energy,power and other industries,CO_(2) emissions rise sharply,which will cause a large amount of CO_(2) in the air be absorbed by the ocean and lead to ocean acidification.The growt...With the rapid development of oil,energy,power and other industries,CO_(2) emissions rise sharply,which will cause a large amount of CO_(2) in the air be absorbed by the ocean and lead to ocean acidification.The growth and development of organisms can be seriously affected by acidified seawater.Sepia esculenta is a mollusk with high nutritional and economic value and is widely cultured in offshore waters of China.Larvae are the early life forms of the organism and are more vulnerable to changes in the external environment.Too low pH will lead to some adverse reactions in larvae,which will affect metabolism,immune response and other life activities.In this study,we sequenced the transcriptome of S.esculenta subjected to acidified seawater stress and identified 1072differentially expressed genes(DEGs).The detected atypical expression of DEGs substantiates cellular malformation and translocation in S.esculenta under low pH stimulation.Simultaneously,this also substantiates the notable impact of ocean acidification on mollusks.These DEGs were used for functional enrichment analysis of GO and KEGG,and the top twenty items of the biological process classification in GO terms and 11 KEGG signaling pathways were significantly enriched.Finally,the constructed proteinprotein interaction network(PPI)was used to analyze protein-protein interactions,and 12 key DEGs and 3 hub genes were identified.The reliability of 12 genes was verified by quantitative RT-PCR.A comprehensive analysis of the KEGG signaling pathway and PPI revealed that ocean acidification leads to abnormalities in lipid metabolism in S.esculenta larvae,which can lead to cancer development and metastasis,accompanied by some degree of inflammation.The results of the study will help to further investigate the physiological processes of S.esculenta when stimulated by ocean acidification,and provide a reference to cope with the captive breeding of S.esculenta affected by acidification.展开更多
Thirty nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH,...Thirty nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample ( P < 0.99 ). The mean values of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017 , while those of the Rizhao sample are 0.023 and 0.025 respectively.展开更多
The secretion function of mantle is closely related to shell formation in some bivalves and gastropods. Up to now, few researches have been reported for cuttlebone formation in the class Cephalopoda. In this study, th...The secretion function of mantle is closely related to shell formation in some bivalves and gastropods. Up to now, few researches have been reported for cuttlebone formation in the class Cephalopoda. In this study, the structure and secretion function of cuttlebone sac of the golden cuttlefish Sepia esculenta was analyzed using the histological and histochemical methods. The results showed that high and columnar cells located in sac epithelium, and fiat cells existed near the base membrane. A lot of fibroblasts were found in the lateral mantle collective tissue. Some mucus, mucopolysaccharide and alkaline phosphatase （ALP） were found in the sac. The ultrastructural characteristics of Quasi-connective-tissue-calcium cells （QCTCC） were observed using a transmission electron microscope （TEM）. The relationship between cuttlebone sac secretion function and shell formation was discussed.展开更多
The morphological characteristics and the cuttlebone formation of Sepia esculenta exposed to different water temperature fluctuations were investigated under laboratory conditions. Temperature fluctuation cycles （15 ...The morphological characteristics and the cuttlebone formation of Sepia esculenta exposed to different water temperature fluctuations were investigated under laboratory conditions. Temperature fluctuation cycles （15 cycles, 60 d in total） consisted of the following three regimes of 4 d duration： keeping water temperature in 26℃ for 3 d （Group A）, 2 d （Group B）, 0 d （Group C, control）; then keeping water temperature in 16℃ for the next 1, 2, 4 d. No significant difference in the survival rate was observed between the control and temperature fluctuation groups （P〉0.05）. Lamellar depositions in a temperature fluctuation cycle were 2.45±0.02 for Group A, 2.00±0.02 for Group B, and 1.78±0.02 for Group C （P〈0.05）. The relationship between age and number of lamellas in the cuttlebone of S. esculenta under each water temperature fluctuation could be described as the linear model and the number of lamellas in the cuttlebone did not correspond to actual age. Group A had the highest cuttlebone growth index （CGI）, the lowest locular index （LI）, and inter-streak distances comparing with those of control group. However, the number of lamellas and LI or CGI showed a quadratic relationship for each temperature fluctuation group. In addition, temperature fluctuations caused the breakage of cuttlebone dark rings, which was considered a thermal mark. The position of the breakage in the dark rings was random. This thermal mark can be used as supplementary information for marking and releasing techniques.展开更多
As a stenohaline species,the survival of Sepia pharaonis can be affected by salinity significantly.This study aimed to explore the function of decreasing salinity on the survival of Sepia pharaonis,which can provide a...As a stenohaline species,the survival of Sepia pharaonis can be affected by salinity significantly.This study aimed to explore the function of decreasing salinity on the survival of Sepia pharaonis,which can provide an advanced production guide on the culture of S.pharaonis in the rainy season.Salinity was gradually decreased from 29 to 22 within 48 h to acclimate S.pharaonis to a low-salinity environment.After ten days of breeding under low-salinity of 22,the death rate was high.In this process,changes in tissue and cell structures in the larval liver,biochemical indicators,and osmoregulation-related gene expression were examined.In-terestingly,hepatocytes in the low-salinity group were irregular,had dissolved tissue inclusions,and contained vacuolized cells.There-fore,low salinity caused severe damages at a cellular level that can elevate the mortality rate.A gradual decline in salinity limited the full adaptation of S.pharaonis.Biochemical indicators and osmoregulation-related gene expression changed similarly.For instance,the trend of malondialdehyde(MAD)as a product of lipid peroxidation reflected the degree of damage to the body by free radicals.The antioxidant system of S.pharaonis could cope with oxidative stress caused by the change in salinity to a certain extent.Osmo-regulation-related genes’expression also showed an optimistic result,that is,S.pharaonis responded positively to the change in sali-nity by adjusting the expression of osmoregulation-related genes.Conversely,the increase in mortality at day 10 also proved the weak adaptation capability of S.pharaonis.This study indicated that S.pharaonis can adapt to a low-salinity environment with a li-mited extent.展开更多
The distribution of chitinolytic enzymes in eight organs of the golden cuttlefish Sepia esculenta was determined. Chitinase activity (activity of endo-type chitinolytic enzyme) was measured using pNP-(GlcNAc)n (n = 2,...The distribution of chitinolytic enzymes in eight organs of the golden cuttlefish Sepia esculenta was determined. Chitinase activity (activity of endo-type chitinolytic enzyme) was measured using pNP-(GlcNAc)n (n = 2, 3) as substrates, with high activity detected in the liver, posterior salivary gland, and stomach. β-N-acetylhexosaminidase (Hex) activity (activity of exo-type chitinolytic enzyme) was determined using pNP-(GlcNAc) as a substrate, and high activity was observed in six organs, including the liver, branchial heart, posterior salivary gland, and stomach. In addition, two chitin-binding proteins (CBP-A, CBP-B) were isolated from the liver using a chitin affinity column. Two full-length cDNAs (SeChi-1: 1484 bp;SeChi-2: 1748 bp) encoding chitinases were obtained from the liver of S. esculenta. SeChi-1 contained a 1377-bp open reading frame (ORF) encoding 459 amino acids, and SeChi-2 contained a 1656-bp ORF encoding 552 amino acids. Domain structures predicted from the deduced amino acid sequences of SeChi-1 and SeChi-2 (SeChi-1, SeChi-2) contained signal peptides, a GH Family 18 catalytic domain, one chitin binding domain (CBD) in SeChi-1, and two CBDs in SeChi-2. Proteome analysis revealed that 125 peptide residues of CBP-A were present in SeChi-1, and 116 peptide residues of CBP-B were present in SeChi-2. Organ expression analysis revealed that SeChi-1 and SeChi-2 were expressed only in the liver of S. esculenta. Phylogenetic analysis of SeChi-1, SeChi-2, and GH family 18 chitinases revealed that SeChi-2 belongs to a group of previously reported squid chitinases, while SeChi-1 does not belong to any previously reported group of mollusk chitinases.展开更多
Diabetes mellitus(DM)has emerged as a serious public health concern,due to the high morbidity and mortality resulted from its complications,such as diabetic nephropathy,diabetic cardiovascular complication,and diabeti...Diabetes mellitus(DM)has emerged as a serious public health concern,due to the high morbidity and mortality resulted from its complications,such as diabetic nephropathy,diabetic cardiovascular complication,and diabetic neuropathy,etc.In this study,we investigated the beneficial effects of sepia ink melanin(SIM)on hyperglycaemia and the restoration of diabetic symptoms in streptozotocin(STZ)-induced diabetic model mice.At first,the normal experimental mice were performed with intraperitoneal injection of STZ(40 mg(kg BW)^(−1))(BW,body weight)to attain diabetes and then were treated with different concentrations of SIM(120,240 and 480 mg(kg BW)^(−1))for four weeks.After treatment,significant decrease in gluconeogenesis were determined,accompanied by a notable increase in both glycolysis and oxidative enzyme activities in SIM-treated groups,such as liver marker enzymes in the serum and key antioxidant enzymes in liver.qPCR results revealed the transcriptional alterations in SIM-treated groups.SIM exposure increased the expression levels of several genes related to insulin transduction and PI3K/Akt pathway,including PI3K,Akt,Irs-2,and InsR.Meanwhile,expression levels of Dicarbonyl/l-xylulose reductase(Dcxr)and UDP-glucose dehydrogenase(Ugdh),which are in-volved in pentose-glucuronate interconversion pathway,were also elevated in SIM-treated groups.Furthermore,histological observation results indicated that nuclear deformation and organelle dissolution were improved,thus could enhance the liver function.These results demonstrated that SIM can be effective in ameliorating diabetic symptoms and improving disease management for diabetic patients.展开更多
基金supported by the National Natural Science Foundation of China(No.31872547)the Natural Science Foundation of Zhejiang Province,China(No.LY20 C190007)+1 种基金the Open Foundation from Marine Sciences in the First-Class Subjects of Zhejiang(No.OFMS004)the Excellent Postdoctoral Program of Jiangsu Province(No.314865).
文摘Neuropeptides are widely distributed in vertebrates and invertebrates,regulating a variety of physiological activities in the organisms,such as metabolism,feeding and reproduction.In this study,to explore the function of neuropeptide LFRFamide in Sepia pharaonis,the full-length cDNA of LFRFamide-like gene(named SpLFRFL,MG869822.1)was identified with rapid amplification of cDNA ends(RACE)method.The sequence of SpLFRFL was 860 bp in length and encoded 188 amino acids containing 4 different mature peptides:1 copy of PHTPFRFamide,NSLFRFamide,TIFRFamide,and 3 copies of GNLFRFamide.Multiple alignment and phylogenetic analysis results showed that SpLFRFL shared high identity with LFRFamides of Sepia officinalis and Sepiella japonica and had the closest relationship with them.Through quantitative Real-time PCR(qRT-PCR),it was found that the SpLFRFL gene was highly expressed in the optic lobe and brain at three different stages during gonad development in both genders.Moreover,the four mature peptides at a concentration of 0.01μmol L^(−1) could inhibit the protein synthesis in the Chinese hamster ovary cell strain-K1(CHOK1)induced by SpGnRH.These data suggest that SpLFRFL might be involved in the development and reproduction of S.pharaonis.The results can contribute to future studies on neuropeptide evolution and function and benefit the cuttlefish farming.
基金funded by the Ministry of Agriculture of the People’s Republic of China (No.CARS-49)。
文摘With the rapid development of oil,energy,power and other industries,CO_(2) emissions rise sharply,which will cause a large amount of CO_(2) in the air be absorbed by the ocean and lead to ocean acidification.The growth and development of organisms can be seriously affected by acidified seawater.Sepia esculenta is a mollusk with high nutritional and economic value and is widely cultured in offshore waters of China.Larvae are the early life forms of the organism and are more vulnerable to changes in the external environment.Too low pH will lead to some adverse reactions in larvae,which will affect metabolism,immune response and other life activities.In this study,we sequenced the transcriptome of S.esculenta subjected to acidified seawater stress and identified 1072differentially expressed genes(DEGs).The detected atypical expression of DEGs substantiates cellular malformation and translocation in S.esculenta under low pH stimulation.Simultaneously,this also substantiates the notable impact of ocean acidification on mollusks.These DEGs were used for functional enrichment analysis of GO and KEGG,and the top twenty items of the biological process classification in GO terms and 11 KEGG signaling pathways were significantly enriched.Finally,the constructed proteinprotein interaction network(PPI)was used to analyze protein-protein interactions,and 12 key DEGs and 3 hub genes were identified.The reliability of 12 genes was verified by quantitative RT-PCR.A comprehensive analysis of the KEGG signaling pathway and PPI revealed that ocean acidification leads to abnormalities in lipid metabolism in S.esculenta larvae,which can lead to cancer development and metastasis,accompanied by some degree of inflammation.The results of the study will help to further investigate the physiological processes of S.esculenta when stimulated by ocean acidification,and provide a reference to cope with the captive breeding of S.esculenta affected by acidification.
文摘Thirty nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample ( P < 0.99 ). The mean values of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017 , while those of the Rizhao sample are 0.023 and 0.025 respectively.
基金This study was supported by the grants from the National Natural Science Foundation of China （No. 30600463）the Key Laboratory of Mariculture of Ministry of Education, 0cean University of China （No. 200610）.
文摘The secretion function of mantle is closely related to shell formation in some bivalves and gastropods. Up to now, few researches have been reported for cuttlebone formation in the class Cephalopoda. In this study, the structure and secretion function of cuttlebone sac of the golden cuttlefish Sepia esculenta was analyzed using the histological and histochemical methods. The results showed that high and columnar cells located in sac epithelium, and fiat cells existed near the base membrane. A lot of fibroblasts were found in the lateral mantle collective tissue. Some mucus, mucopolysaccharide and alkaline phosphatase （ALP） were found in the sac. The ultrastructural characteristics of Quasi-connective-tissue-calcium cells （QCTCC） were observed using a transmission electron microscope （TEM）. The relationship between cuttlebone sac secretion function and shell formation was discussed.
基金Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2010AA10A404)the National Marine Public Welfare Research Project (No. 200805069)the NMOE Project (No. 1011010603)
文摘The morphological characteristics and the cuttlebone formation of Sepia esculenta exposed to different water temperature fluctuations were investigated under laboratory conditions. Temperature fluctuation cycles （15 cycles, 60 d in total） consisted of the following three regimes of 4 d duration： keeping water temperature in 26℃ for 3 d （Group A）, 2 d （Group B）, 0 d （Group C, control）; then keeping water temperature in 16℃ for the next 1, 2, 4 d. No significant difference in the survival rate was observed between the control and temperature fluctuation groups （P〉0.05）. Lamellar depositions in a temperature fluctuation cycle were 2.45±0.02 for Group A, 2.00±0.02 for Group B, and 1.78±0.02 for Group C （P〈0.05）. The relationship between age and number of lamellas in the cuttlebone of S. esculenta under each water temperature fluctuation could be described as the linear model and the number of lamellas in the cuttlebone did not correspond to actual age. Group A had the highest cuttlebone growth index （CGI）, the lowest locular index （LI）, and inter-streak distances comparing with those of control group. However, the number of lamellas and LI or CGI showed a quadratic relationship for each temperature fluctuation group. In addition, temperature fluctuations caused the breakage of cuttlebone dark rings, which was considered a thermal mark. The position of the breakage in the dark rings was random. This thermal mark can be used as supplementary information for marking and releasing techniques.
基金supported by the Ningbo Agricultural Major Projects (No. 201401C1111001)the Foundation of Zhejiang Educational Committee (No. Y2019409 57)sponsored by K.C.Wong Magna Fund in Ningbo University
文摘As a stenohaline species,the survival of Sepia pharaonis can be affected by salinity significantly.This study aimed to explore the function of decreasing salinity on the survival of Sepia pharaonis,which can provide an advanced production guide on the culture of S.pharaonis in the rainy season.Salinity was gradually decreased from 29 to 22 within 48 h to acclimate S.pharaonis to a low-salinity environment.After ten days of breeding under low-salinity of 22,the death rate was high.In this process,changes in tissue and cell structures in the larval liver,biochemical indicators,and osmoregulation-related gene expression were examined.In-terestingly,hepatocytes in the low-salinity group were irregular,had dissolved tissue inclusions,and contained vacuolized cells.There-fore,low salinity caused severe damages at a cellular level that can elevate the mortality rate.A gradual decline in salinity limited the full adaptation of S.pharaonis.Biochemical indicators and osmoregulation-related gene expression changed similarly.For instance,the trend of malondialdehyde(MAD)as a product of lipid peroxidation reflected the degree of damage to the body by free radicals.The antioxidant system of S.pharaonis could cope with oxidative stress caused by the change in salinity to a certain extent.Osmo-regulation-related genes’expression also showed an optimistic result,that is,S.pharaonis responded positively to the change in sali-nity by adjusting the expression of osmoregulation-related genes.Conversely,the increase in mortality at day 10 also proved the weak adaptation capability of S.pharaonis.This study indicated that S.pharaonis can adapt to a low-salinity environment with a li-mited extent.
文摘The distribution of chitinolytic enzymes in eight organs of the golden cuttlefish Sepia esculenta was determined. Chitinase activity (activity of endo-type chitinolytic enzyme) was measured using pNP-(GlcNAc)n (n = 2, 3) as substrates, with high activity detected in the liver, posterior salivary gland, and stomach. β-N-acetylhexosaminidase (Hex) activity (activity of exo-type chitinolytic enzyme) was determined using pNP-(GlcNAc) as a substrate, and high activity was observed in six organs, including the liver, branchial heart, posterior salivary gland, and stomach. In addition, two chitin-binding proteins (CBP-A, CBP-B) were isolated from the liver using a chitin affinity column. Two full-length cDNAs (SeChi-1: 1484 bp;SeChi-2: 1748 bp) encoding chitinases were obtained from the liver of S. esculenta. SeChi-1 contained a 1377-bp open reading frame (ORF) encoding 459 amino acids, and SeChi-2 contained a 1656-bp ORF encoding 552 amino acids. Domain structures predicted from the deduced amino acid sequences of SeChi-1 and SeChi-2 (SeChi-1, SeChi-2) contained signal peptides, a GH Family 18 catalytic domain, one chitin binding domain (CBD) in SeChi-1, and two CBDs in SeChi-2. Proteome analysis revealed that 125 peptide residues of CBP-A were present in SeChi-1, and 116 peptide residues of CBP-B were present in SeChi-2. Organ expression analysis revealed that SeChi-1 and SeChi-2 were expressed only in the liver of S. esculenta. Phylogenetic analysis of SeChi-1, SeChi-2, and GH family 18 chitinases revealed that SeChi-2 belongs to a group of previously reported squid chitinases, while SeChi-1 does not belong to any previously reported group of mollusk chitinases.
基金supported by the Natural Science Foundation of Zhejiang Province(No.LY18C190006)sponsored by K.C.Wong Magna Fund in Ningbo University.
文摘Diabetes mellitus(DM)has emerged as a serious public health concern,due to the high morbidity and mortality resulted from its complications,such as diabetic nephropathy,diabetic cardiovascular complication,and diabetic neuropathy,etc.In this study,we investigated the beneficial effects of sepia ink melanin(SIM)on hyperglycaemia and the restoration of diabetic symptoms in streptozotocin(STZ)-induced diabetic model mice.At first,the normal experimental mice were performed with intraperitoneal injection of STZ(40 mg(kg BW)^(−1))(BW,body weight)to attain diabetes and then were treated with different concentrations of SIM(120,240 and 480 mg(kg BW)^(−1))for four weeks.After treatment,significant decrease in gluconeogenesis were determined,accompanied by a notable increase in both glycolysis and oxidative enzyme activities in SIM-treated groups,such as liver marker enzymes in the serum and key antioxidant enzymes in liver.qPCR results revealed the transcriptional alterations in SIM-treated groups.SIM exposure increased the expression levels of several genes related to insulin transduction and PI3K/Akt pathway,including PI3K,Akt,Irs-2,and InsR.Meanwhile,expression levels of Dicarbonyl/l-xylulose reductase(Dcxr)and UDP-glucose dehydrogenase(Ugdh),which are in-volved in pentose-glucuronate interconversion pathway,were also elevated in SIM-treated groups.Furthermore,histological observation results indicated that nuclear deformation and organelle dissolution were improved,thus could enhance the liver function.These results demonstrated that SIM can be effective in ameliorating diabetic symptoms and improving disease management for diabetic patients.
