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ABSTRACT: Annual changes in the gonadosomatic index (GSI), oocyte diameter, gonadal histology and plasma estradiol-17β (E2 ) levels were investigated in female common Japanese conger Conger myriaster in captivity. Juveniles were caught in September 1999 and reared in seawater at temperatures ranging from 10–20°C for 3 years. All fish were immature when captured in September 1999. GSI and oocyte diameter increased from October 2000, peaked between summer and autumn 2001, and bottomed-out in winter 2001. Plasma E2 level also increased from October 2000, but remained high. The ovarian developmental stage was at the peri-nucleolus stage or the oil droplet stage until September 2000. Vitellogenesis started in October 2000 and oocytes progressed to the tertiary yolk globule stage by summer 2001. However, vitellogenic oocytes regressed in all females after autumn 2001. The neogenetic oocytes began to increase after November 2001 and ovarian development progressed in 2002 as they did in 2001, although maximum GSI in 2002 was half its 2001 value. These data indicate that ovarian development in the common Japanese conger has an annual periodicity, and that these congers may be able to spawn in multiple years under rearing condition. 相似文献
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ABSTRACT: Interleukin-1β (IL-1β) cDNA of Japanese flounder was found to consist of 1329 bp, encoded 247 amino acid residues. Among the fish IL-1β in the databases, the one with the highest identity of Japanese flounder IL-1β was that of seabass (62% identity). The expression of IL-1β was induced by treatment with concanavalin A (ConA)/phorbol myristate acetate (PMA) and lipopolysaccharide. The copy number of IL-1β mRNA was increased 30-fold after stimulation with ConA/PMA. Of 871 cDNA on a microarray, 93 genes (10.7%) were up-regulated or down-regulated by IL-1β at 1, 3 and 7 days post-injection. The induced gene expression was highest on day 1 followed by day 3 and day 7. A total of 7% of known and 3.7% of unknown genes of the 871 tested genes were differentially expressed. Of the genes tested, 7.4% were up-regulated and 3.3% were down-regulated. Cytokine genes such as tumor necrosis factor, granulocyte colony stimulating factor and chemokine receptor A were induced in response to IL-1β. Cell surface antigens such as IgM, MHC class I and CD20 receptor were up-regulated. Signal transduction genes such as Toll-like receptor 1 and SH3P2 were also up-regulated. The glucocorticoid receptor and cAMP early repressor were down-regulated in our microarray analysis. 相似文献
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Seiko Nonaka Tomonori Somamoto Yoko Kato-Unoki Mitsuru Ototake Teruyuki Nakanishi Miki Nakao 《Fisheries Science》2008,74(2):341-346
ABSTRACT: The clonal triploid ginbuna crucian carp Carassius auratus langsdorfii , a naturally occurring gynogenetic fish, is a useful model for studying T-cell-mediated immunity. CD4, a T-cell receptor (TCR) coreceptor, is a membrane-bound glycoprotein found on helper T-cells, and assists in the binding of major histocompatibility complexes. In the present study, full-length cDNAs encoding the CD4 molecule from the S3n strain of ginbuna crucian carp were cloned and characterized. 5'-rapid amplification of cDNA ends (RACE) and 3'-RACE yielded two distinct cDNA clones of CD4 homolog from the ginbuna, and these sequences share 95% identity at the amino acid level. These ginbuna CD4 molecules consisted of a signal peptide, immunoglobulin superfamily (IgSf) like domains, a transmembrane domain, and a cytoplasmic domain similar to other known CD4. A tyrosine protein kinase p56lck binding motif is conserved in the cytoplasmic tail of ginbuna CD4. Phylogenetic tree analysis indicated that ginbuna CD4 sequences are closely related to CD4L-1 from other fish species. Expression of ginbuna CD4 mRNA was detected in the gill, thymus, head kidney, trunk kidney and peripheral blood leukocytes, indicating that its expression pattern is similar to that of ginbuna TCRβ mRNA. The results suggest that ginbuna CD4 sequences are useful as molecular probes for helper T-cells. 相似文献
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Yan Tan Kiyoshi Osatomi Yukinori Nozaki Tadashi Ishihara Kenji Hara 《Fisheries Science》2006,72(1):185-194
ABSTRACT: We purified cathepsins B1 and B2 from the ordinary muscle of carp Cyprinus carpio . The N-terminal amino acid sequences (12 residues) of 29 kDa bands of cathepsins B1 and B2 are the same and showed high homology of 75% and 83%, respectively, with the heavy chain of rat and human cathepsins B. Based on conserved sequences of other cathepsins B and the N-terminal amino acid sequences of 29 kDa bands, we cloned carp cathepsin B cDNA. The nucleotide sequence of carp cathepsin B cDNA consists of 1470 bp including a 993 bp open reading frame, encoding a deduced protein of 330 amino acids. The deduced amino acid sequence of carp cathepsin B has similarity of 80% to rainbow trout cathepsin B and of 76–78% to other vertebrate cathepsins B. The sequence of its isoform was also determined during molecular cloning, which has 94.8% similarity with first cloned cathepsin B. They are completely same in N-terminal amino acid sequence of heavy chain, active site and potential N-glycosylation site. This indicates there are at least two kinds of cathepsin B functioning in vivo in carp. 相似文献
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ABSTRACT: The nucleotide sequences of Japanese flounder Paralichthys olivaceus , major histocompatibility complex (MHC) cDNA, classical MHC class Iα, non-classical MHC class Iβ, MHC class IIα and IIβ, were determined. The domain structures and antigen binding motifs of vertebrate MHC are conserved in the Japanese flounder MHC. A phylogenetic analysis supports the classification of these genes into class I and class II MHC. Classical MHC class Iα was ubiquitously expressed, whereas the non-classical MHC class Iβ was expressed mainly in lymphoid organs, gills, intestine and stomach. The MHC classes IIα and IIβ were also ubiquitously expressed. 相似文献
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Masatomi Hosoi Chuya Shinzato Masaya Takagi Shoko Hosoi-Tanabe Hideki Sawada Eri Terasawa Haruhiko Toyohara 《Fisheries Science》2007,73(2):385-394
ABSTRACT: Taurine is the primary osmolyte in marine molluscs, whose cellular osmo-conforming process is vital for environmental adaptation because of a lack of osmotic homeostasis. Here, cDNA cloning and expression, and functional analyses of taurine transporter (TAUT) from the giant Pacific oyster are reported on. The deduced amino-acid sequence of oyster TAUT (oyTAUT) showed 47–51% identity to those of vertebrate TAUT, whereas identity among the vertebrates is 78–95%. Functional analysis of oyTAUT expressed in Xenopus oocytes revealed that oyTAUT has a lower affinity and specificity for taurine and a requirement for higher NaCl concentration, compared with vertebrate TAUT. Taken together with similar functional properties of TAUT from mussel, indicated by our previous study, it is possible that these functional features reflect the internal environment of the molluscs (i.e. higher taurine and NaCl concentrations). Oyster taurine transporter mRNA expression was induced by not only hyper-osmotic stress, similar to other TAUT, but also hypo-osmotic stress. It is speculated that the expression in response to hypo-osmotic stress was induced by a substantial decrease in tissue taurine content following the decrease in the internal osmolality. 相似文献
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Alberto Teodorico Correia Rui Faria Paulo Alexandrino Carlos Antunes Eduardo José Isidro João Coimbra 《Fisheries Science》2006,72(1):20-27
ABSTRACT: The European conger eel Conger conger is an important marine benthic fish in the North-East Atlantic and represents a valuable fishery resource. However, little is known about its reproductive biology. In an attempt to gain a better understanding of the conger eel population structure, mitochondrial DNA (mtDNA) sequences were examined. A region with 432 bp of the control region of the mtDNA was sequenced from 40 individuals from six different locations around the central and eastern North Atlantic Ocean. Thirty variable positions defined 28 distinct haplotypes. The average sequence difference within samples (1.3–4.2%) was comparable to those between samples (1.4–3.6%). MtDNA sequence-based statistical tests showed significant geographic differentiation between some local population samples, suggesting that the conger eel does not comprise a single panmictic population. However, given our sample sizes, these preliminary results should be interpreted with caution and more individuals from more sites, including the Mediterranean Sea, should be analyzed in detail. The genetic variability detected in this study is an initial step to elucidate the genetic background of the conger eel population structure. 相似文献
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Cloning of kuruma prawn Marsupenaeus japonicus crustin-like peptide cDNA and analysis of its expression 总被引：1，自引：0，他引：1
Achara RATTANACHAI Ikuo HIRONO Tsuyoshi OHIRA Yukinori TAKAHASHI Takashi AOKI 《Fisheries Science》2004,70(5):765-771
ABSTRACT: Antimicrobial peptides serve as an important component of the innate immune system of all species by functioning to provide a rapid first line defense against infection. Arthropod antimicrobial peptides have been well described in insects, whereas only a few molecules have been identified in crustaceans. Five variants (types 1–5) of Marsupenaeus japonicus crustin-like peptide cDNA that were obtained from a hemocyte cDNA library and polymerase chain reaction (PCR) amplification are reported here. Marsupenaeus japonicus crustin-like peptide type 1, the predominant type, has a cDNA consisting of 679 nucleotides and an open reading frame consisting of 573 base pairs coding for 191 amino acid residues. Other types contain varying glycine-rich repeats at the N-terminal amino acid sequences. The deduced amino acid sequences of these variants are highly similar to those of Litopenaeus setiferus (80% identity), Litopenaeus vannamei (80% identity) and Carcinus maenas crustins (44% identity). Expression of Marsupenaeus japonicus crustin-like peptide mRNA was detected in hemocytes, but not in the heart, hepatopancreas, gill, fore-gut, mid-gut, muscle, subcuticular epithelium or ovary. The expression level of crustin-like peptide mRNA increased significantly 1, 3 and 7 days post-peptidoglycan feeding as determined by quantitative real-time PCR. These results suggest that crustin-like peptide could have an important role in shrimp defense mechanisms. 相似文献
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ABSTRACT: In the present study, the cDNA encoding myoglobin (Mb) of bigeye tuna Thunnus obesus was cloned and its amino acid sequence deduced in order to investigate the relationship between the primary structure and thermostability of scombridae fish Mb. An open reading frame of bigeye tuna Mb cDNA contained 444 nucleotides encoding 147 amino acids. The primary structure of bigeye tuna Mb was highly conserved when compared with those of bluefin tuna and yellowfin tuna Mb, the sequence identity being 95.2–100.0%. It also showed relatively high identity (82.3–89.1%) with the counterparts of scombridae fish. Myoglobin was then isolated from the dark muscle of four scombridae fish including bigeye tuna. Differential scanning calorimetry and circular dichroism measurements on these Mb revealed that the thermostability of bigeye tuna Mb was lowest and that of skipjack Katsuwonus pelamis Mb highest among the scombridae fish Mb examined. The α-helical contents of scombridae fish Mb at 10°C were in the range of 39.8–44.8%, clearly lower than that of horse Mb (55.3%), suggesting instability of fish Mb. The melting temperatures of these Mb fell in the range of 75.7–79.9°C, lower than that of horse Mb (84.2°C). These results strongly suggest the instability of fish Mb. 相似文献
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雌激素是重要的类固醇类激素,在软体动物中,雌激素一般不能与雌激素受体识别并结合,但却能通过调控一类孤儿核受体——雌激素相关受体(estrogen-related receptor,ERR)的表达而发挥其生殖调控的部分功能,但有关ERR在软体动物中的确切生物学特性及功能还知之甚少。本研究首次从2种海洋头足类:曼氏无针乌贼(Sepiella japonica)和白斑乌贼(Sepia latimanus)中,克隆得到了ERR基因的全序列,并对其在乌贼不同组织及不同发育时期的表达特征进行研究。结果表明:乌贼中仅克隆得到了1种ERR基因亚型(ERRγ),在曼氏无针乌贼和白斑乌贼中的全长分别为1 513 bp和1 547 bp,两者分别编码339和351个氨基酸残基,这些氨基酸可形成A/B、C、D和E 4个典型的核受体结构域。进化树分析表明,乌贼的ERR基因C结构域和E结构域较其他软体动物相对高度保守,曼氏无针乌贼和白斑乌贼ERR基因与同为头足类的双斑蛸(Octopus bimaculoides)的ERR同源性最高,分别达到95%和85%。2种乌贼ERR基因在不同组织中表达谱相仿,在不同组织中广泛表达,但在脑、肝、卵巢等生殖相关器官中表达量最高。2种乌贼卵巢中的ERR基因表达量都随性成熟时间的推进而逐渐上调,而在繁殖期后显著下调(P<0.05)。该结果表明乌贼的ERR基因可能在其生殖发育和性成熟过程中起着重要的调控作用。[中国渔业质量与标准,2019,9(2):16-23] 相似文献
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Structural and thermodynamic characterization of tropomyosin from fast skeletal muscle of bluefin tuna 总被引：1，自引：0，他引：1
ABSTRACT: Tuna tropomyosin is a mixture of nearly equimolar amounts of two isoforms (designated α and β). cDNA encoding the α form was cloned from bluefin tuna Thunnus thynnus fast skeletal muscle. The full-length cDNA contained 1220 bp, comprising an open reading frame of 855 bp encoding 284 amino acid residues, flanked by 5'-untranslational regions (156 bp) and 3'-untranslational regions (209 bp). The deduced amino acid sequence showed considerably high homology in a range of 93.7–98.6% to those of other vertebrate α-type tropomyosins. In phylogenetic analysis, bluefin tuna tropomyosin showed the closest relationship with the white croaker counterpart. The predicted mass was 32 919 Da, and isoelectric point was 4.50, assuming acetylation of the N-terminus. By differential scanning calorimetry, bluefin tuna tropomyosin gave two major endothermic peaks at 29.3 and 41.5°C, probably caused by the presence of two isoforms. Circular dichroism spectra supported such a unique denaturation profile. 相似文献
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NORIYUKI HORIE TOMOKO UTOH YOSHIAKI YAMADA AKIHIRO OKAMURA HUAN ZHANG NAOMI MIKAWA ATSUSHI AKAZAWA SATORU TANAKA HIDEO P OKA 《Fisheries Science》2002,68(5):972-983
Development of embryos and larvae in the common Japanese conger Conger myriaster was observed after artificial fertilization. Eggs were obtained from females matured artificially by hormone injections and milt was obtained from males matured naturally. Fertilized eggs were kept in seawater at 12–14°C. The first cleavage occurred at 4 h, epiboly began at 24 h, the embryonic body was formed at 38 h and hatching occurred at 84 h after insemination. Newly hatched larvae were approximately 2.5 mm (total length) and similar to those of Anguilla japonica in terms of external features. The mouth and anus opened on the 7th day after hatching. Pigments began to appear at the tip of the tail on the 10th day. The total length of the larvae reached approximately 8 mm on the 11th day. Eye pigmentation began on the 14th day. One larva lived for 19 days without food. 相似文献
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Previously we isolated the major alginate lyase isozyme LbAly35 from a marine snail Littorina brevicula and showed that this enzyme was significantly heat stable in a broad pH range compared with other molluscan alginate lyases (Hata et al., Fish Sci 75:755?C763, 2009). LbAly35 showed practically no similarity to other molluscan alginate lyases in the N-terminal amino-acid sequence of 20 residues and no cross-reactivity with anti-abalone alginate lyase antiserum. These led us to consider that the primary structure of LbAly35 is considerably deviated from other molluscan enzymes. Thus, in the present study, we first compared the thermal stability of LbAly35 with an abalone alginate lyase, HdAly, and found that the first order inactivation rate constants for LbAly35 at 40 and 45?°C were 1/20 and 1/45 of those for HdAly, respectively. Then, we cloned cDNAs encoding LbAly35 and characterized its deduced amino-acid sequence comparing with those of other molluscan alginate lyases. The cDNAs were amplified by polymerase chain reaction (PCR) and 5??- and 3??-RACE PCRs from the L. brevicula hepatopancreas cDNA using degenerated primers synthesized on the basis of partial amino-acid sequences of LbAly35. The cDNA covering the entire translational region of LbAly35 comprised 1,093?bp and encoded an amino-acid sequence of 296 residues. The amino-acid sequence consisted of an initiation methionine, a putative signal peptide for secretion (22 residues), a propeptide-like region (10 residues), and a mature LbAly35 domain of 263 residues. Although the N-terminal region of LbAly35 was significantly deviated from those of other molluscan alginate lyases, the catalytic domain of LbAly35 showed ~45?% identity to other molluscan enzymes which had been classified under polysaccharide-lyase-family-14 (PL-14). In addition, the amino-acid residues crucially important for the catalytic actions of PL-14 enzymes were also conserved in LbAly35. Accordingly, LbAly35 was regarded as a member of PL-14 as other molluscan alginate lyases despite the significant deviation of its N-terminal region. 相似文献
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Dongdong Han Huijun Miao Qin Nie Shuyan Miao Qin Zhang Wenbing Zhang Kangsen Mai 《Fish physiology and biochemistry》2016,42(6):1665-1679
In the present study, the full-length cDNA sequences of leptin (LEP) and its receptor (LEPR) from turbot Scophthalmus maximus were cloned. The cDNA of tLEP was 1126 bp in length encoding 157 amino acids. The amino acid sequence shared low identity with human LEP (18.8 %), but the three-dimensional structures of these two LEPs were strongly conserved. The deduced 1173-amino acid sequence of tLEPR was 28 % identical to human LEPR, and 82 % too range-spotted grouper LEPR, containing all functionally important domains conserved in vertebrate LEPR. Tissue distribution analysis showed that tLEP was abundantly expressed in brain, eyes and liver. The highest level of tLEPR mRNA was found in liver and kidney. After a 9-week feeding trial using diets with different ratios of carbohydrate–lipid (1:6, 1:2, 2:1 and 14:1), it was found that the increase in dietary carbohydrate-to-lipid ratios from 1:6 to 2:1 did not significantly influence tLEP and tLEPR expression in turbot liver (P > 0.05). The hepatic tLEP expression was significantly elevated in treatment with 14:1 dietary carbohydrate-to-lipid ratio (P < 0.05). The hepatic tLEPR mRNA level in group with 14:1 dietary carbohydrate-to-lipid ratio was significantly lower than that in 1:6 group (P < 0.05), but had no significant difference with the other two groups (P > 0.05). These results revealed the important relationship between dietary carbohydrate-to-lipid ratio and LEP expression in turbot. 相似文献
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Victor S Panangala Riccardo Russo Vicky L van Santen Karen G Wolfe & Phillip H Klesius 《Aquaculture Research》2009,40(10):1135-1147
Edwardsiella ictaluri , the cause of enteric septicaemia in channel catfish ( Ictalurus punctatus ), is motile by means of peritrichous flagella. We determined the complete flagellin gene sequences and their organization in E. ictaluri by sequencing genomic segments from a λ-ZAP phage genomic library of E. ictaluri . Four flagellin genes ( fliC1, fliC2, fliC3 and fliC4 ) are arranged in tandem within 6 kb in the E. ictaluri genome. Each flagellin-coding sequence is preceded by a σ28 recognition site consensus sequence. The predicted amino acid sequences of all four flagellin proteins (between 36 and 37.5 kDa) are similar in the N-terminal (1–160 aa) and C-terminal (last 74 aa) portions and are divergent in the central portion of the proteins. Proteins encoded by flC1, fliC2 and fliC3 are more similar to each other (88–90% aa identity) than to the protein encoded by fliC4 (76–78% aa identity). basic local alignment search tool analysis of GenBank sequences showed that all flagellin aa sequences are more similar to those of Serratia marcescens (72–74% identity) than to those of Edwardsiella tarda (≤64% identity). Primary determination of E. ictaluri flagellin gene sequences facilitate advanced studies on the role of flagella in host–pathogen interaction. 相似文献
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为进一步了解鱼类MHC ⅡA基因的特点及其在免疫反应中的功能,采用同源克隆、RACE-PCR、巢式PCR等技术,从健康的尼罗罗非鱼体获得1 205 bp的MHC ⅡA基因cDNA全序列(Orni-DBA-0101,Genebank登录号:JF719813)及1 388 bp的基因组序列。序列分析发现,尼罗罗非鱼MHC ⅡA基因含4个外显子和3个内含子,开放阅读框长720 bp,编码239个氨基酸。从4尾尼罗罗非鱼中共得到8条不同的cDNA序列,分别编码不同的氨基酸序列。氨基酸序列比对后发现,序列间存在丰富的多态性,且主要集中在α-1区,多态性位点数远远高于半滑舌鳎MHC ⅡA基因。生物信息学分析表明,尼罗罗非鱼MHC ⅡA编码的蛋白质分子包含1个信号肽、2个胞外结构域、1个跨膜区和1个胞质区,存在4个保守的半胱氨酸残基以及丰富的磷酸化位点,与其他物种的相似性为23%～65%。RT-PCR结果表明,MHC ⅡA基因在脾、肾、肠、鳃、性腺、肝、心脏表达量很高,在鳔和肌肉中表达量最低。人工感染嗜水气单胞菌后,肝、脾、肾、鳃、肠5个组织中MHC ⅡA基因的mRNA水平均发生了不同程度的变化,提示MHC ⅡA分子作为一种重要的免疫因子,在清除病原的免疫反应中起着重要作用。 相似文献