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1.
Corticotropin-releasing hormone (CRH) is a potent mediator of endocrine, autonomic, behavioral, and immune responses to stress. For a better understanding of the structure and function of the CRH gene and to study its effect on feeding regulation in cyprinid fish, the cDNA of the CRH gene from the brain of Schizothorax prenanti was cloned and sequenced. The full-length CRH cDNA consisted of 1,046 bp with an open reading frame of 489 bp encoding a protein of 162 amino acids. Real-time quantitative PCR analyses revealed that CRH was widely expressed in central and peripheral tissues. In particular, high expression level of CRH was detected in brain. Furthermore, CRH mRNA expression was examined in different brain regions, especially high in hypothalamus. In addition, there was no significant change in CRH mRNA expression in fed group compared with the fasted group in the S. prenanti hypothalamus during short-term fasting. However, CRH gene expression presented significant decrease in the hypothalamus in fasted group compared with the fed group (P < 0.05) on day 7; thereafter, re-feeding could lead to a significant increase in CRH mRNA expression in fasted group on day 9. The results suggest that the CRH may play a critical role in feeding regulation in S. prenanti.  相似文献   

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3.
We examined the polyploidy of Cyprinus carpio, the German mirror carp. Specimens were collected in the field in Hulan, China, and treated with phytohemagglutinin and colchicine before the chromosome spreads and the karyotype of kidney cells were examined using silver staining, chromomycin A3 (CMA3)/distamycin (DA)/4,6-diamidino-2-phenylindole (DAPI) staining, and fluorescence in situ hybridization (FISH) with a 5.8S + 28S rDNA probe. One to twosilver stained (Ag) nucleolus organizing regions (NORs) were detected during metaphase and interphase events, with 80 % of the metaphase spreads and 69 % of the interphase nuclei showing two Ag-NORs signals. One CMA3 signal was detected on the terminus of the short arm of each submetacentric chromosome 8 (SMC8) homolog (n = 2). The 5.8S + 28S rDNA probe hybridized at the terminus of the short arm of each SMC8 homolog (n = 2). The results of the Ag-NORs, CMA3/DA/DAPI, and 5.8S + 28S rDNA FISH analyses were consistent with regard to the total number and location of the SMC8 NORs in the chromosome spreads and karyotype, indicating that, at the molecular cytogenetic level, the German mirror carp is an evolutionary tetraploid with two sets of chromosomes after diploidization.  相似文献   

4.
Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt’s lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.  相似文献   

5.
Glycoconjugates are known to be involved in many physiological events in vertebrates. Sialidase is one of the glycosidases, which removes sialic acid from glycoconjugates. In mammals, the properties and physiological functions of sialidases have been investigated, while there is little understanding of fish sialidase. Here, to investigate the significance of fish neu4 sialidase, neu4 gene was cloned from medaka brain mRNA and identified. Sialidase-specific motifs (GPG, YRVP and Asp-Box) were well conserved in the medaka neu4 polypeptide. Optimal pH of medaka neu4 sialidase was 4.6, but its activity was sustained even at neutral and weak alkaline pH. The neu4 considerably cleaved sialic acid from 4-methylumbelliferyl-N-acetyl-α-d-neuraminic acid and sialyllactose, but not from ganglioside and fetuin, which are good substrates for human NEU4. neu4 activity was mostly detected in mitochondria/lysosome fraction after biochemical fractionation, and indirect immunofluorescence assays revealed neu4 localization in lysosome in neu4 overexpressed cells. Next, developmental change in medaka neu4 and other sialidase mRNA levels were estimated by real-time PCR. Each sialidases showed different expression patterns in embryonic development: neu4 was up-regulated at late developmental stage in embryo, and neu3a mRNA level was quite high in 0.5 dpf. On the other hand, neu3b expression was drastically increased after hatching, suggesting that each sialidase may play a different role in embryonic development.  相似文献   

6.
The trace element contents of Corbicula clam shells collected from Japan, Russia, China, and the Republic of Korea were analyzed to determine their geographic origin. The crushed shells were decomposed with nitric acid–hydrogen peroxide, and the concentrations of 14 elements (Li, Mg, V, Mn, Co, As, Rb, Mo, Ba, Ce, Pb, U, Sr, and Ca) were measured by inductively coupled plasma mass spectrometry and inductively coupled plasma optical emission spectrometry. Some of the elements identified in samples displayed a geographic trend. The average content of manganese in Japanese samples was twice that of Russian samples. Conversely, the arsenic content in Japanese samples was approximately half of that in Russian samples. Linear discriminant analysis was applied to the data from Japanese and Russian samples, and a discriminant model was constructed. The discriminant model was used to determine the geographic origin of Corbicula clams produced in Japan, with 89.8 % of those identified as Japanese and 92.2 % of those identified as Russian being classified correctly. Therefore, trace element analysis of the shells of Corbicula clams is a useful technique for the identification of their country of origin.  相似文献   

7.
Oxidative stress causes damage at the cellular level and activates a number of signaling pathways. Heat-shock proteins (HSPs) play an important role in repair and protective mechanisms under cell response to stress conditions. HSP70 has been shown to act as an inhibitor of apoptosis. Apoptosis signal-regulating kinase-1 (ASK1) activity is regulated at multiple levels, one of which is through inhibition by cytosolic chaperons HSP70. The current study was aimed to investigate the alteration in signaling molecules that allow the fish to survive under stressed natural field conditions. The study also investigates the variation in biomolecular composition of hepatocytes by using Fourier transform infrared spectroscopy. The impact of stress on hepatocytes was assessed by measuring the level of lipid peroxides (LPO), catalase activity (CAT) and assessing the changes in hepatocytes of Mugil cephalus inhabiting Kovalam and Ennore estuaries. The expression of HSP70 and ASK1 were analyzed by immunoblot analysis and ELISA, respectively. The spectral analysis showed variations in biomolecular composition of hepatocytes at a wave number region of 4,000–400 cm?1. There was significant decrease of CAT activity (p < 0.01) (25 %) with significant increase of LPO (p < 0.001) (35 %) and HSP70 (p < 0.001) and insignificant increase of ASK1 (p < 0.05) (16 %) in fish hepatocytes inhabiting Ennore estuary than Kovalam estuary. In conclusion, the present study suggests that the survival of fish in the Ennore estuary under stressed condition may be due to the upregulation of HSP70 that mediates the altered signal pathway which promotes cellular resistance against apoptosis.  相似文献   

8.
Abalone Haliotis discus hannai Ino is a popular delicacy consumed as a luxury food owing to its unique flavor and texture. In this study, we investigated the effects of heating conditions on the fatty acids and volatile compounds in its foot muscle based on gas chromatography–mass spectrometry (GC–MS) and solid-phase microextraction (SPME)–GC–MS. The contents of fatty acids significantly decreased after heating at 80 °C for 2 h. In total, 52 volatile compounds, including aldehydes, aromatic compounds, alkanes, alcohols, ketones, and furans, were detected in the heated samples. Principal component analysis revealed an interaction between heating temperature and time. Heating at 80 °C for 0.5–2 h generated higher contents of volatile compounds. In particular, the contents of hexanal, heptanal, octanal, nonanal, and undecanal mainly derived from autoxidation of fatty acids during heating increased at least fourfold.  相似文献   

9.
The role of environmental ion composition and osmolality in Ca2+ signaled activation was assessed in spermatozoa of brook trout Salvelinus fontinalis. Milt from ten mature males was obtained by abdominal massage. Spermatozoa motility was evaluated in 0, 100, and 300 mOsm/kg NaCl or sucrose solutions, buffered by 10 mM Tris–HCl pH 8.5. For investigation of spermatozoa reaction to external Ca2+ concentration, 2 mM ethylene glycol tetraacetic acid (EGTA) was added to the activation media as a calcium ions chelator. For investigation of the effect of external Na+ concentration in conditions of low external Ca2+, 100 µM amiloride was added to the EGTA-containing solutions as a Na+ transport blocker. Low motility was observed in sucrose (Na+ free) solutions containing 2 mM EGTA but not in Na+ solutions containing 2 mM EGTA. Addition of amiloride led to significantly increased motility (P < 0.05) compared with sucrose (Na+ free) solutions containing 2 mM EGTA. We conclude that Na+ transport in Ca2+-free solutions plays a regulatory role in brook trout spermatozoa activation. The influence of competitive Na+ and Ca2+ transport on the control of spermatozoa activation requires further study with respect to its application for improvement of artificial activation and storage media.  相似文献   

10.
Fish gills are the vital multifunctional organ in direct contact with external environment. Therefore, activation of the cytoprotective mechanisms to maintain branchial cell viability is important for fish upon stresses. Salinity is one of the major factors strongly affecting cellular and organismal functions. Reduction of ambient salinity may occur in coral reef and leads to osmotic stress for reef-associated stenohaline fish. However, the physiological responses to salinity stress in reef-associated fish were not examined substantially. With this regard, the physiological parameters and the responses of protein quality control (PQC) and osmoregulatory mechanisms in gills of seawater (SW; 33–35 ‰)- and brackish water (BW; 20 ‰)-acclimated blue-green damselfish (Chromis viridis) were explored. The results showed that the examined physiological parameters were maintained within certain physiological ranges in C. viridis acclimated to different salinities. In PQC mechanism, expression of heat-shock protein (HSP) 90, 70, and 60 elevated in response to BW acclimation while the levels of ubiquitin-conjugated proteins were similar between the two groups. Thus, it was presumed that upregulation of HSPs was sufficient to prevent the accumulation of aggregated proteins for maintaining the protein quality and viability of gill cells when C. viridis were acclimated to BW. Moreover, gill Na+/K+-ATPase expression and protein amounts of basolaterally located Na+/K+/2Cl? cotransporter were higher in SW fish than in BW fish. Taken together, this study showed that the cytoprotective and osmoregulatory mechanisms of blue-green damselfish were functionally activated and modulated to withstand the challenge of reduction in salinity for maintaining physiological homeostasis.  相似文献   

11.
In this study, full-length tilapia transferrin (OnTF) isolated from liver cDNA of Nile tilapia (Oreochromis niloticus) was found to have an open reading frame of 2,091-bp encoding 696 amino acid residues. Two additional amino acids: Gly369 and Gly370 were observed compared with the reported Nile tilapia transferrin protein sequence. Pre-mature protein has a predicted molecular weight of 78.2 kDa, while mature protein is 73.28 kDa in size. Comparative sequence analysis with transferrin from other species revealed two major putative iron-binding domains designated as the N-lobe and the C-lobe in accordance with the transferrin protein characteristics. The predicted tertiary structure of tilapia transferrin confirmed the presence of iron and anion-binding sites on both lobes that are conserved among transferrins from other species. Quantitative real-time PCR analysis showed significantly higher expression of tilapia transferrin gene in liver than in other tissues (p < 0.05). Transferrin expression in tilapia experimentally infected with 106 and 108 colony-forming units mL?1 of Streptococcus agalactiae was significantly upregulated at 24 and 12 h post-infection (hpi), respectively, and decreased afterward. Iron-deficiency in serum of bacterially infected fish was detected at 48 and 24 hpi, respectively. The expression pattern of the transferrin gene and the iron levels of infected tilapia in this study were consistent with the function of transferrin in innate immunity.  相似文献   

12.
In the present study, a full-length cDNA encoding the insulin-like growth factor binding protein-2 (IGFBP-2) was cloned from the liver of goldfish (Carassius auratus) by rapid amplification of cDNA ends technique. The goldfish IGFBP-2 cDNA sequence was 1,513 bp long and had an open reading frame of 825 bp encoding a predicted polypeptide of 274 amino acid residues. Semi-quantitative RT-PCR results revealed that goldfish IGFBP-2 mRNA was expressed in all detected tissues. In liver, central nervous system and pituitary gland, goldfish IGFBP-2 expressed at high levels, followed by anterior intestine, middle intestine and kidney. In posterior intestine, ovary, skin, fat, spleen, muscle and gill, the goldfish IGFBP-2 expression levels were very low. Fasting and refeeding experiment showed that the mRNA expression of goldfish IGFBP-2 was up-regulated significantly in liver compared to the fed group and restored rapidly to normal level after refed. However, the mRNA expressions of IGFBP-2 in hypothalamus and pituitary of goldfish were insensitive to fasting. Furthermore, the mRNA expressions of IGFBP-2 in hypothalamus, pituitary and liver were varied in periprandial changes and significantly down-regulated at 2 and 4 h after meal. These results imply that the IGFBP-2 mRNA expression may be associated with anabolic and catabolic metabolism and regulated by metabolic factors in goldfish.  相似文献   

13.
The luteinizing hormone receptor (LHR) plays a crucial role in female reproduction. In the present study, full-length sequence coding for the LHR was obtained from female turbot (Scophthalmus maximus) by homology cloning and a strategy based on rapid amplification of cDNA end-polymerase chain reaction. The full-length LHR cDNA was 3,184 bp long and contained a 2,058-bp open reading frame which encoded a protein of 685 amino acids. Multiple sequence alignments of the turbot LHR manifested high homologies with the corresponding sequences of available teleosts and representative vertebrates, and significant homology with that of Hippoglossus hippoglossus. In addition, the turbot LHR showed typical characteristics of glycoprotein receptors, including a long N-terminal extracellular domain, seven transmembrane domains, and a short C-terminal intracellular domain. LHR mRNA was abundant in the ovary, but was deficient in extra-ovarian tissues. Furthermore, LHR mRNA gradually developed from previtellogenesis to migratory nucleus stage, with the highest values observed in migratory nucleus stage during reproductive cycle. However, LHR mRNA sharply decreased in atresia stage. These results suggested that LHR is a typical G protein-coupled receptor that is involved in the promotion of turbot ovarian development and may be related to the final maturation and ovulation of oocyte. These findings contribute to the understanding of the potential roles of LHR in controlling the fish reproductive cycle.  相似文献   

14.
The immunomodulatory effect of garlic–mineral oil as a modified adjuvant was investigated in rohu, Labeo rohita along with formalin-killed Aeromonas hydrophila. The garlic extract was not having any proteolytic effect on the tested antigen as marked in SDS-PAGE. The efficacy of graded levels of modified adjuvant formulation was assessed by measuring serum hemolysin and agglutination titers as well as level of protection against A. hydrophila challenge in the immunized fish at days 10 and 30 post-vaccination (dpv). The higher hemolysin and bacterial agglutinin titers along with significantly lower mortality against A. hydrophila challenge in modified adjuvant-based vaccinated fish, particularly at low level of garlic incorporation, indicated the pronounced immunoadjuvant role of this modified adjuvant formulation in rohu.  相似文献   

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17.
The recent increase of the local population of gilthead seabream, Sparus aurata, in three areas along the southeastern Adriatic Sea: Malostonski Bay (Croatia and Bosnia and Herzegovina), Neretva Estuary (Croatia) and Boka Kotorska Bay (Montenegro) and its adverse effects on shellfish culture by preying on Mediterranean mussels, Mytilus galloprovincialis, and the European flat oyster, Ostrea edulis, are studied. The results from the analysis of the existing information show that the main reason for the recent increase is the escapes from local fish farm which enrich the local population constantly with new gilthead sea bream. The existence of practically endless food in the area of the shellfish farms allows the concentration of the population in the region instead of its dispersion along the Adriatic coast. Moreover, ecological analysis indicates that the gilthead seabream is facing a very low competition from other local species which enhances its capacity to further populate the region. While the impact on the ecosystem is not yet known, the socio-economic impact of the increase of the gilthead seabream population is evident today. Many shellfish farms are closing today in the region since the damages may reach over 90 % of the production.  相似文献   

18.
The mollusc Pinctada margaritifera is the top economic aquaculture species in French Polynesia (export value) and forms the basis of the black pearl industry. Mass production of this unique gem, produced by a living organism, relies on a surgical operation requiring tissue from a donor pearl oyster to be grafted, together with a nucleus made of shell, into the gonad of a receiving pearl oyster. This technique is performed by expert grafters, whose work constitutes the first step influencing pearl farm production yield. This study makes the first report of effects mediated by individual grafters and by the season of grafting on three rates scored at 45 days post operation: nucleus retention, nucleus rejection and pearl oyster mortality. These results were obtained in a very large scale grafting experiment, designed and conducted in a single culture site in the atoll of Arutua (Tuamotu Archipelago) and involving a total of 52,910 grafts performed by ten professional grafters, during two contrasting seasons: autumn (four grafting experiments) and spring (three grafting experiments). Statistical analysis using linear mixed-effect model for both univariate and multivariate analysis was performed. The results show a very highly significant season effect (p < 0.001), with a higher rate of nucleus retention in autumn than spring: 91.1 versus 79.4 %, respectively. A very highly significant season effect was also recorded for nucleus rejection rate (p < 0.001), which was greater in spring than in autumn: 12.6 versus 7.7 %. Oyster mortality rate was up to six times higher in spring than in autumn (p < 0.001). Within the two seasons, there was no significant difference between the grafting experiments for any of the three variables, adding robustness to the inter-season results. Taking into account this seasonal variation effect, grafter skill had a significant impact on the three variables (p < 0.001), making it possible to rank grafters from most to least efficient. Aside from skill differences between grafters, grafting in autumn may significantly increase graft success. Water temperature and its indirect consequences could be one of the most important environmental factors implicated in overall efficiency. These findings may help the potential development of an annual grafting schedule according to season/ lagoon temperature as a way to maximize production yield for the black-lipped pearl oyster industry in French Polynesia.  相似文献   

19.
Fish are commonly exposed to environmental pollutants, which in turns could induce an oxidative stress. So, it is important to understand the effects and the responses elicited by these toxicants in fish species, being fish cell lines important tools for this purpose. Thus, the aim of the present study was to compare the effects of copper and UV-B radiation exposure on zebrafish hepatocytes (ZFL lineage) in terms of reactive oxygen species (ROS) levels, sulfhydril groups content and mRNA levels of important genes related to cellular response to toxic agents. Exposure of ZFL cells to UV-B radiation (23.3 mJ/cm2) significantly increased levels of intracellular ROS and mRNA of both superoxide dismutase isoforms (sod1 and sod2), three glutathione S-transferase isoforms (gstα, gstµ and gstπ) and a heat shock protein (hsp70). However, no changes in nonprotein sulfhydryl groups (NP-SH) content, as well as in the mRNA levels of genes related to glutathione (GSH) synthesis and recycling, were observed. Contrary to this, copper exposure (20 mg/L) diminished NP-SH content and increased the levels of mRNA of genes related to GSH synthesis (gclc and gs). Moreover, copper exposure increases the mRNA levels of some genes related to antioxidant defenses (gpx and gstπ), biotransformation reactions (cyp1a1) and protein repair (hsp70). In conclusion, these results demonstrated that both toxicants could increase ROS levels in ZFL cell line, but the responses are different, which could be related to activation of different signaling pathways.  相似文献   

20.
The honmoroko has been inferred to have an XX/XY sex determination system, but the parental genome can also affect the sex ratio of the offspring. The extent of parental effects on sex determination was examined by checking the sex ratios of F1 and F2 gynogenetic diploids and control diploids. Eleven gynogenetic broods from different females consisted of all or nearly all females, but eight broods showed a variable proportion of males (<50 %). One second-generation brood of gynogenetic diploids consisted wholly of females, but others produced some males. In crosses with a control diploid female, four males from a high-percentage male brood of gynogenetic diploids produced offspring with a balanced sex ratio. Sib-mating between a gynogenetic female and three gynogenetic males from the brood produced predominantly male progeny. These results suggest that there are at least four possible genotypes: genotypic female (XX), phenotypic female carrying a silent Y chromosome, genotypic male (XY), and genotypic supermale (YY). These inferences suggest that this fish has an XY system but a relatively high proportion of females possess a mutated, silent Y chromosome which does not lead to testis formation.  相似文献   

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