Molecular characterization and subcellular localization of Carassius auratus interferon regulatory factor-1

Interferon (IFN)-regulatory transcription factor-1 (IRF-1) has been studied in mammals and fish but little is known about the relationship between its gene structure and nuclear 'ion of IRF-1 protein. In this study, a cDNA encoding Carassius auratus IRF-1 (CaIRF-1) was isolated from an interferon-producing cell line, C. ouratus blastulae embryonic (CAB) cells, exposed to UV-inactivated grass carp hemorrhagic virus (GCHV). The CaIRF-1 genomic locus exhibits exon-intron arrangements similar to those of other vertebrate IRF-1 loci, with nine exons and eight introns, although together with pufferfish IRF-1, CaIRF-1 distinguishes itself from other vertebrate IRF-1 genes by a relatively compact genomic size. Similar to the known IRF-1 genes, CaIRF-1 is ubiquitously expressed, and is upregulated in vitro and in vivo in response to virus, Poty I:C, or CAB INF-containing supernatant (ICS). Subcellular localization analysis confirms the nuclear distribution of CaIRF-1 protein, and reveals two nuclear localization signals (NILS), any one of which is sufficient for nuclear translocation of CaIRF-1. One NLS Locates to amino acids 117-146, and appears to be the structural and functional equivalent of the NLS in mammalian IRF-1. The second NLS (amino acids 73-115) is found within the DNA-binding domain (DBD) of CaIRF-1, and contains two regions rich in basic amino acids (''(KDKSINK101)-K-95" and ''(75)KTWKANFR(82)"). In comparison with mammalian IRF-1, in which the corresponding amino acid stretch does not seem to drive nuclear translocation, five conserved basic amino acids (K-75, K-78, R-82, K-95, and K-101) and one non-conserved basic amino acid (K-97) are present in this NLS from CaIRF-1. This observation suggests that K97 Of CaIRF-1 might be essential for the function of its second NLS, wherein the six basic aminoacids might cooperate to drive CaIRF-1 to the nucleus. Therefore, the current study has revealed a new nuclear localization motif in the DBD of a vertebrate IRF-1. (C) 2007 Elsevier Ltd. All rights reserved.

Identification of genome organization in the unusual allotetraploid form of Carassius auratus gibelio

The unusual allotetraploid form with unequal contribution of chromosome sets was discovered from the gynogenetic offspring of Carassius auratus gibelio stimulated by red common carp sperm. In this study, genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH) with 45S rDNA probe are used. The GISH results lead to the identification of species-specific chromosomes, which permits to demonstrate the origin and genome organization in the allotetraploid form. Moreover, chromosome localization of 45S rDNA and co-localizations of 45S rDNA and Cyprinus carpio genomic DNA further confirm that one extra 45S rDNA positive chromosome in the allotetraploid form originates from the paternal haploid genome of C carpio, and other 5 45S rDNA-containing chromosomes are from the maternal genome of Carassius auratus gibelio. And, the correlation between 45 rDNA and the nucleolar organizer regions (NORs) is confirmed by silver nitrate staining. The data provide direct experiment evidence that the allotetraploid actually contains three chromosome sets of Carassius auratus gibelio and one chromosome set of C carpio, and will be a useful genetic material for both basic research and breeding practice. (c) 2006 Elsevier B.V. All rights reserved.

The use of the ethanol pathway in goldfish Carassius auratus (L.) following anoxia

Goldfish (Carassius auratus) were subjected, for a period of 6 weeks, to 2h progressive hypoxia followed by 6h anoxia in closed respirometers at 15 degree C. The concentrations of glucose, lactate and ethanol were determined in whole goldfish following exposure to both hypoxia and anoxia. Lactate accumulation (mmol/kg/h) was 0.35 during the 1st week but declined to 0.14 in the 6th week of exposure to anoxia. In contrast, ethanol excreted to the surrounding water, increased from 65% to 92% of the total production in the lst and 6th week, respectively. The switch from lactate accumulation to ethanol pathway utilization, with the resultant metabolic depression and anoxia resistance is discussed

Efecto de la temperatura y la ración sobre el metabolismo de la carpa dorada (Carassius auratus)

[ES] Se llevan a cabo tres experimentos para estudiar el efecto de tres factores el metabolismo de Carassius auratus: temperatura de exposición, ración de alimento diario y tiempo transcurrido tras la exposición a las nuevas condiciones.

Probiotics and nutrient supplement in artificial feed of gold fish (Carassius auratus)

The efficacy of three feeds incorporated with different probiotics -Lactobacil, Vizylac and Cyfolac as nutrient supplement was evaluated in an ornamental fish, Carassius auratus (Linn.). The basal diet (40% protein) was prepared and the probiotics were incorporated at different levels viz. Lactobacil at 8x10 super(7)/100g and 12x10 super(7)/100g, Vizylac at 8x10 super(7)/100g and 12x10 super(7)/100g and Cyfolac at 12x10 super(7)/100g. Feeding trial was conducted for a period of 4 weeks. At the end of the experiment, bio-growth parameters and proximate composition of the fishes were studied. The growth, survival and protein content were improved in all the probiotic fed fishes compared to control. The maximum growth (0.74 g) and survival (85%) were observed in fishes fed with Lactobacil at 12x10 super(7)/100g. It emphasizes that supplementation of feed with probiotics has a positive impact on the growth, survival and the body composition of goldfish.

Effect of feeding bioencapsulated Lactobacillus sp. in live Tubifex sp. on the growth performance of gold fish Carassius auratus (Linnaeus, 1758)

An attempt was made to feed bioencapsulate Lactobacillus sp. in live fish food organism Tubifex for use in the culture of gold fish Carassius auratus. The C. auratus fries when fed with bioencapsulated Lactobacillus sp. in Tubifex showed significant improvement in total wet weight gain (p<0.007) and FCR (p<0.01) compared to control. The specific growth rale and mean survival were slightly higher, although insignificantly (p>0.05) in bioencapsulated Tubifex fed group. None of the bacteriological parameters of the fish gut between the experimental and control groups differed significantly (p>0.05). Lactobacillus sp. was recorded at a level of log 5.11/g on the 90th day of experimentation. When the experimental C. auratus fries were infected with Pseudomonas fluorescents, the bioencapsulated Tubifex fed group resisted the infection. The survival was significantly higher (p<0.05) in bioencapsulated Tubifex fed group (44%) than in control (22%). The C. auratus fed with bioencapsulated Tubifex showed less (55%) signs of tail/fin rot. Likewise, a significant improvement in total wet weight gain (p<0.009), FCR (p<0.01) and SGR (p<0.04) of C. auratus brooder fed with bioencapsulated Tubifex was seen compared to control group fed with depurated Tubifex.

粪便收集时间和饲料中肉骨粉含量对异育银鲫(Carassius auratus gibelio)消化率的影响(英文)

本研究探讨了通过收集器不同粪便收集时间和饲料中肉骨粉( MBM)含量对异育银鲫干物质、蛋白、能量、磷的表观消化率(ADC)影响。不同梯度(0 ,20 ,40 ,60 ,80 ,100 %)的肉骨粉替代鱼粉(FM)蛋白配制成六种等氮(粗蛋白:410 g/kg)等能(总能:18 kJ/g)的饲料,通过11周的饲养实验,实验开始后2周开始收集粪便,收集时间分别是:排粪后1 min,投喂后4h和16h。结果表明,干物质、蛋白、能量、磷的消化率明显随粪便收集时间增加而升高(p<0 .05) ,但在高肉骨粉替代饲料中

由鲤(Cyprinus carpio)和鲫(Carassius auratus)染色体组叠加构建的两个单性人工多倍体克隆

具有天然雌核发育的多倍体杂种鱼可防止杂种优势的分离并保持其后代的杂种优势。由于假设诱发的多倍体鱼类的生殖模式是天然雌核发育的,我们进行了鲤鲫杂种的多倍体诱发,目的是描述经染色体组叠加由有性鲤鲫二倍体转化为异源三倍体及异源四倍体克隆谱系。鲤鲫杂种产生未减数而具有两亲本染色体组杂种卵子,未减数的雌核可与入卵的雄核融合叠加形成三倍体合子。鲤鲫异源三倍体胚胎发育正常,部分异源三倍体雌性个体可产生未减数的、仍保留母本的三套染色体的成熟卵子。绝大部分鲤鲫异源人工三倍体个体的成熟卵子的雌核不与入卵的雄核融合,具有天然雌

Cytological Mechanism on the Integration of Heterologous Genome or Chromosomes in the Unique Gynogenetic Carassius auratus gibelio

银鲫（ＣａｒａｓｉｕｓａｕｒａｔｕｓｇｉｂｅｌｉｏＢｌｏｃｈ）是行天然雌核发育生殖的两性型三倍体鱼类，与普通两性融合生殖鱼类相比，具有独特的育种优势。八十年代以来，异育银鲫、复合四倍体异育银鲫的发现表明，雌核发育卵子不但具有保持自身全部染色体的能力，还能整合异源精子的部分遗传物质或整个基因组，影响雌核发育后代的性状。因此，搞清楚异源基因组的整合机制对于进一步弄清其发育模式以及诱导复合多倍体银鲫均具有十分重要的作用。两性融合发育鱼类的精子入卵后，精核在促精核活化因子的诱导下，可以逐渐解凝并形成雄性原核；而在

Identification and characterization of hypoxia-induced genes in Carassius auratus blastulae embryonic cells using suppression subtractive hybridization

Organisms living in water are inevitably exposed to periods of hypoxia. Environmental hypoxia has been an important stressor having manifold effects on aquatic life. Many fish species have evolved behavioral, physiological, biochemical and molecular adaptations that enable them to cope with hypoxia. However, the molecular mechanisms of hypoxia tolerance in fish, remain unknown. in this study, we used suppression subtractive hybridization to examine the differential gene expression in CAB cells (Carassius auratus blastulae embryonic cells) exposed to hypoxia for 24 h. We isolated 2100 clones and identified 211 differentially expressed genes (e-value <= 5e-3; Identity > 45%). Among the genes whose expression is modified in cells, a vast majority involved in metabolism, signal transduction, cell defense, angiogenesis, cell growth and proliferation. Twelve genes encoding for ERO1-L, p53, CPO, HO-1, MKP2, PFK-2, cystatin B, GLUT1, BTG1, TGF beta 1, PGAM1, hypothetical protein F1508, were selected and identified to be hypoxia-induced using semi-quantitive RT-PCR and real-time PCR. Among the identified genes, two open reading frames (ORFs) encoding for CaBTG1 and Cacystatin B were obtained. The deduced amino acid sequence of CaBTG1 had 94.1%, 72.8%, 72.8%, 72.8%, 68.6% identity with that of DrBTG1, HsBTG1, BtBTG1, MmBTG1 and XIBTG1. Comparison of Cacystatin B with known cystatin B, the molecules exhibited 49.5 to 76.0% identity overall. These results may provide significant information for further understanding of the adaptive mechanism by which C. auratus responds to hypoxia. (c) 2008 Elsevier Inc. All rights reserved.

The effect of cyanobacterial crude extract on the transcription of GST mu, GST kappa and GST rho in different organs of goldfish (Carassius auratus)

The glutathione S-transferases play important roles in the detoxification of microcystin. Core-sequences of three classes of GST (mu, kappa and rho) were cloned from goldfish (Carassius auratus L) i.p. injected with cyanobacterial crude extract at two doses (50 and 200 mu g MC-LReq kg(-1) BW). The relative changes of the mRNA abundance in liver, kidney and intestine were analyzed by real-time PCR. The transcription of GST mu was inhibited in intestine at both doses and the transcription of GST kappa was inhibited from 12 to 48 h in kidney at both doses. The decreased transcription of GST rho was detected in all three organs at the high dose. It is suggested that transcription inhibition of GST rho might be significant in MCs toxicity at higher toxin concentration in omnivorous freshwater fish. Alteration in transcription of GSTs stimulated by MCs implicates an increased health risk to fish. (C) 2008 Published by Elsevier B.V.

Identification of a C1q family member associated with cortical granules and follicular cell apoptosis in Carassius auratus gibelio

C1q family proteins with C1q domain have been reported in vertebrates, but their biological roles are currently unknown. In this study, a C1q-like factor, designated Carassius auratus gibelio ovary-specific C1q-like factor (CagOC1q-like), was identified as a cortical granules component. Immunofluorescence localization revealed that the C1q family member was specifically expressed in follicular epithelial cells, and associated with cortical granules in fully grown oocytes. Moreover, it was discharged to the perivitelline space and egg envelope upon fertilization. As it is the first identified C1q family member that is expressed in follicular cells that surround oocyte, CagOC1q-like was applied to detection of follicular cell apoptosis and deletion. The entire cytological process of follicular cell apoptosis and deletion was clearly seen from double visualizations of follicular cells with CagOC1q-like immunofluorescence and apoptotic follicular cells labeled by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) during oocyte maturation and ovulation. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Clonal diversity and genealogical relationships of gibel carp in four hatcheries

To conserve and utilize the genetic pool of gynogenetic gibel carp (Carassius auratus gibelio), the Fangzheng and Qihe stock hatcheries have been established in China. However, little information is available on the amount of genetic variation within and between these populations. In this study, clonal diversity in 101 fish from these two stock hatcheries and 35 fish from two other hatcheries in Wuhan and Pengze respectively was analysed for variation in serum transferrin. Thirteen clones were found in Fangzheng and Qihe, of which 12 were novel. Six clones were specific to Fangzheng and three specific to Qihe, whereas four were shared among the Fangzheng and Qihe fish. To obtain more knowledge on genetic diversity and genealogical relationships within gibel carp, the complete mitochondrial DNA (mtDNA) control region (similar to 920 bp) was sequenced in 64 individuals representing all 14 clones identified in the four hatcheries. Differences in the mtDNA sequences varied remarkably among hatcheries, with the Fangzheng and Qihe lines demonstrating high diversity and Wuhan and Pengze showing no variation. The Fangzheng and Qihe lines might represent two distinct matrilineal sources. One of the Qihe samples carried the haplotype shared by a most widely cultivated Fangzheng clone, indicating that a Fangzheng clone escaped from cultivated ponds and moved into the Qihe hatchery. Four Fangzheng samples clustered within the lineage formed mainly by Qihe samples, most likely reflecting historical gene flow from Qihe to Fangzheng. It is suggested that clones in Wuhan originated from Fangzheng, consistent with their introduction history, supporting the hypothesis that gibel carp in Pengze were domesticated from individuals in the Fangzheng hatchery.