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Ruilong Wang Yun Gao Lihong Zhang Yike Zhang Zhanqiang Fang Jianguo He Weimin Zhang Guangzhi Ma 《Fish physiology and biochemistry》2010,36(2):157-164
Vitellogenins (Vtgs), the precursors for the yolk proteins, are very important for the embryonic development of teleosts,
and have also been studied extensively as biomarkers for environmental estrogenic mimics. The cDNA for a Vtg was isolated
from the liver of the female white cloud mountain minnow (Tanichthys albonubes) by 3′- and 5′-RACE methods. It is 4,171 bp in full length, and encodes a putative protein of 1,326 amino acids. This putative
Vtg, designated as wcmmVtg, contains complete portions of LVI and PV, but lacks the C-terminal half of LVII and thus belongs
to type I vitellogenin. In addition to the liver of the female fish, wcmmVtg was also shown to be expressed in the ovary.
During ovarian development, the mRNA expression of wcmmVtg in both the liver and ovary was continuously increased from the
previtellogenic to late vitellogenic stages, but then decreased significantly at post-spawning stage. In the male fish, expression
of wcmmVtg mRNA was induced in the liver by treatment with E2 (10 and 100 ng/l) for 14 days. These results suggest that the Vtg originated from the ovary of the white cloud mountain minnow
may also contribute to the accumulation of yolk proteins during oocyte growth, and that the male white cloud mountain minnow
is sensitive to the estrogenic treatment in terms of Vtg mRNA expression, which could also be applied to monitor the environmental
estrogenic mimics. 相似文献
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Fisheries Science - The article "Development of an efficient bioreactor system for delivering foreign proteins secreted from liver into eggs with a vitellogenin signal in medaka Oryzias... 相似文献
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F. Chu-Koo R. Dugué M. Alván Aguilar A. Casanova Daza F. Alcántara Bocanegra C. Chávez Veintemilla F. Duponchelle J.-F. Renno Salvador Tello J. Nuñez 《Fish physiology and biochemistry》2009,35(1):125-136
Arapaima gigas is an air-breathing giant fish of Amazonian rivers. Given its great economic and cultural importance, the aquaculture development
of this species represents an evident solution to face the decline of wild populations. In captivity, reproduction occurs
generally in large earthen ponds where stocks of a few tens of brooders are maintained together at the beginning of the rainy
season (December–March in the Peruvian Amazon). Fry production relies on the spontaneous formation of male and female pairs,
which build a nest, delimit a territory and guard the offspring for at least 20 days from other congeners and predators. However,
as sex determination of A. gigas is not possible by morphological criteria, it is very difficult to optimize reproduction conditions and fry production in
each pond, which seriously hampers the culture of this species. This situation prompted us to develop sexing methodologies
based on (1) the detection of female specific plasma Vitellogenin (Vtg) using an enzyme immuno assay (EIA), and (2) the determination
of plasma 17β-estradiol and 11-ketotestosterone levels for immature specimens. The Vtg purification was performed by electro-elution
after polyacrilamide gel electrophoresis (PAGE) from plasma of 17β-estradiol treated A. gigas juveniles. Two different Vtg molecules were isolated, (Vtg1 and Vtg2) with 184 and 112 kDa apparent molecular masses, respectively, and two antibodies were raised in rabbits for each Vtg molecule.
Adult fish were 100% accurately sexed by Vtg EIA, while 100% of immature fish and 95% of adults were accurately sexed by 17β-Estradiol
and 11-Ketestosterone ratios. We also observed different color pattern development in male and female adult fish (6-year-olds)
around the reproductive period. 相似文献
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In vitro effect of pituitary,interrenal and gonadal hormones on vitellogenin synthesis in primary hepatocyte cultures of Chalcalburnus tarichi 
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下载免费PDF全文 Vitellogenin (Vtg) is an important precursor yolk protein in egg‐laying vertebrates, including fish. The 17β‐oestradiol (E2) plays a crucial role in the Vtg synthesis; moreover, certain hormones can stimulate Vtg synthesis. We investigated the possible role of E2, carp recombinant growth hormone (crGH), insulin (Ins), progesterone (P4) and 11‐deoxycortisol (11‐DOC) hormones in Vtg synthesis on primary juvenile Chalcalburnus tarichi hepatocyte culture. The amount of Vtg in the medium was measured at 2‐day intervals using enzyme‐linked immunosorbent assay (ELISA). The hepatocytes were maintained in culture for more than 2 weeks without the addition of serum components. Vitellogenin localization was visualized with the immunofluorescence method in E2‐supplemented hepatocytes. Among hormones applied to the culture, only E2 had an influence on Vtg synthesis in a time‐dependent manner, while crGH, Ins, P4 and 11‐DOC had no effect. However, in hepatocytes stimulated with E2 in combination with P4, a lower Vtg production was seen compared with Vtg produced when hepatocytes were stimulated with E2 alone. P4 proved to have potentiating effects on co‐treatment with E2‐induced Vtg production. As a result, E2 and P4 are the most important hormones for Vtg synthesis in juvenile C. tarichi hepatocyte culture. 相似文献
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D. Manohar G. Damodar Rao G. Sreenivasulu B. Senthilkumaran Aparna Dutta Gupta 《Fish physiology and biochemistry》2005,31(2-3):235-239
Vitellogenin (Vtg) is a female specific glycophospholipoprotein which can be induced both in male and female with estradiol
and xeno-estrogens. The basic theme behind the purification of vitellogenin from the fish is to understand the evolutionary
relationship and for the purification and characterization of the Vtg receptor. The male catfish, Clarias gariepinus was administrated with estradiol over a period of time for the synthesis of Vtg and the serum was collected. The Vtg was
purified from the serum using a two step chromatographic technique. The serum was passed on to DEAE-ion exchange column and
the protein was eluted using a salt gradient. The fractions containing the Vtg were pooled and passed onto a gel permeation
chromatography column and the pure protein was obtained. The molecular weight is around 200 kDa on the SDS-PAGE and around
520 kDa on the native gel electrophoresis. 相似文献
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Theodore I. J. Smith Edward K. Dingley Donald E. Marchette 《Journal of the World Aquaculture Society》1981,12(2):78-87
In South Carolina, a joint state-federal program was initiated in 1977 to develop spawning and rearing techniques for Atlantic sturgeon, Acipenser oxyrhynchus, and to investigate its culture potential. Good progress has been made in developing the hatchery technology. Brood stock collection sites and techniques have been established and suitable transportation and holding procedures demonstrated for these large fish (? 40–100 kg). Homospecific hypophyseal injections were employed successfully to induce two spawnings. The artificially fertilized eggs, incubated in MacDonald hatching jars, hatched in about 120 hours at 17–18°C. Yolk sac was abosrbed by day 10 and the fry were weaned on a diet consisting of beef liver and salmon mash supplemented with live Artemia. During the second trial, fish hatched at 7.8 mm mean total length obtained a size of 176.7 mm in 204 days. Several pond culture studies were undertaken with juvenile sturgeon. Although most fish eventually died (due to poor environmental conditions) some wild-caught juveniles were successfully maintained under pond conditions for about 20 months and one brood female survived for 15 months. Pond-held fish were fed trout chow supplemented with pieces of beef liver. Feeding activity could be observed at night along dike areas. 相似文献
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Sachi Kume Naoto Katayama Kensuke Ichida Shoko Hattori-Ihara Kazue Nagasawa Goro Yoshizaki 《Fisheries Science》2014,80(4):767-773
Cre/loxP-mediated cell targeting is considered to be a powerful tool for biotechnology in farmed fish. As a first step toward establishing cell targeting in salmonids, we analyzed the functionality of the Cre/loxP system in rainbow trout. We first established stable transgenic strains carrying the DsRed gene, which was flanked by loxP sites and further spliced with the EGFP gene. By microinjecting Cre complementary RNA (cRNA) into fertilized eggs of the transgenic trout, the functionality of the Cre/loxP system was evaluated. The results showed that all of the embryos exhibited green fluorescence in at least some of their cells. While 19 out of 20 embryos comprised cells showing both green and red fluorescence, the remaining embryo showed only green fluorescence. Polymerase chain reaction (PCR) using primers designed to recognize sequences outside of the two loxP sites revealed that, in addition to long intact fragments, the 19 individuals carried short fragments that were equivalent in length to the loxP-excised fragments. The remaining green embryo carried only this short fragment. DNA sequencing of the short fragment revealed that it lacked the DNA fragments flanking the loxP sites and the spliced fragments did not contain any sequence rearrangements. These results suggest that the Cre/loxP system is functional in rainbow trout. 相似文献
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Shunmei Liu Xiaonan Zang Bin Liu Xuecheng Zhang Arunakumara K K I U Xiaoqing Zhang & Bing Liang 《Aquaculture Research》2007,38(12):1283-1292
The present study investigated the effect of supplementing feed with transgenic Synechocystis sp. PCC6803 containing the Paralichthys olivaceus growth hormone (GH) gene on growth, feed intake and feed efficiency ratio, muscle composition, haematology and histology of turbot (Scophthalmus maximus L.). At the end of the 40‐day feeding trial, the specific growth rate of fish fed the supplemented feed with 1.0% transgenic Synechocystis sp. PCC6803 was 21.67% higher (P<0.05) than that of control fish. Although body weight and feed efficiency ratio significantly increased (P<0.05) in fish fed the diet supplemented with transgenic alga, feed intake and condition factor of the experimental fish were unaffected. Muscle composition analysis showed that the protein content was positively influenced by the transgenic alga, whereas the lipid content was unaffected. Haematological parameters, including red blood cell, white blood cell, haemoglobin, and serum biochemical indices, such as enzyme activities of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, concentrations of total protein, glucose, blood urea nitrogen, creatinine, triglyceride and cholesterol and ion levels of K, Na, Cl, P were not influenced by supplementing the transgenic Synechocystis sp. PCC6803. Furthermore, no histopathological alterations were induced by transgenic alga treatment in the stomach, intestine, liver, spleen and kidney of the experimental fish. The results of the present study indicated that transgenic Synechocystis sp. PCC6803 containing P. olivaceus GH gene is an efficient growth promoter and a safe feed additive for fish. 相似文献
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Vitellogenin (Vtg) was purified from the plasma of 17-estradiol (E2)-injected male greenback flounder,Rhombosolea tapirina. The molecular weight of the native Vtg was estimated by gel filtration as 540 kD. SDS-PAGE and Western blotting analyses indicated that this protein consisted of three bands with molecular weights of 155, 104, 79 kD, respectively. A polyclonal antibody against the highest molecular weight band of putative Vtg was generated in sheep and an indirect antibody-capture competitive enzyme-linked immunosorbent assay (ELISA) was developed. The assay was validatedfor plasma Vtg measurement in greenback flounder. Serial dilutions of plasma from vitellogenic females parallelled the standard Vtg curve, whereas no cross-reaction was observed with the plasma of males in the ELISA. The Vtg ELISA was used to assess the induction of Vtg by E2
in vivo in males. The induction of Vtg in greenback flounder showed a time- and dose-dependent response as in other species. In E2-treated fish, detectable levels of Vtg were first found at 48 h, and reached a peak at 96 h post-injection. Plasma levels of Vtg increased as the E2 dose increased with a threshold of 0.1 mg kg–1. 相似文献
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Carp (Cyprinus carpio) lipovitellin is a highly stable phospholipoglycoprotein with the same immunogenicity as vitellogenin 下载免费PDF全文
下载免费PDF全文 Jun Wang Zhenzhong Zhang Yanyan Wei Mingyi Zheng Shaoguo Ru 《Aquaculture Research》2018,49(4):1389-1395
Vitellogenin (Vtg) induction in carp (Cyprinus carpio) is a commonly used biomarker for oestrogenic contamination. However, the accurate quantification of Vtg was challenged because the easy degradation of Vtg standard would change the standard curves of the immunoassays. In this study, three yolk proteins were purified from carp ovarian extracts by a two‐step chromatographic method. The purified proteins were characterized as phospholipoglycoproteins with molecular weights of approximately 416, 398 and 383 kDa. In SDS‐PAGE, the purified proteins appeared as a major band of approximately 110 kDa and several faint bands. Based on these characteristics, purified proteins were identified as carp lipovitellin (Lv). Immunological analysis showed that anti‐Vtg antiserum reacted with the purified Lv. The results of stability analysis revealed that even heated at 60°C for 60 min, the electrophoretic patterns of carp Lv in native‐PAGE and SDS‐PAGE did not have a significant difference compared with the Lv solution stored at 4°C. Therefore, the purified carp Lv was confirmed to have similar immunogenicity with Vtg and possess exceptionally high stability, which would be helpful for the development of robust immunoassays for accurate quantification of carp Vtg. 相似文献
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V. N. Williams B. J. Reading N. Hiramatsu H. Amano N. Glassbrook A. Hara C. V. Sullivan 《Fish physiology and biochemistry》2014,40(2):395-415
The multiple vitellogenin (Vtg) system of striped bass, a perciform species spawning nearly neutrally buoyant eggs in freshwater, was investigated. Vitellogenin cDNA cloning, Western blotting of yolk proteins (YPs) using Vtg and YP type-specific antisera, and tandem mass spectrometry (MS/MS) of the YPs revealed the complex mechanisms of yolk formation and maturation in this species. It was discovered that striped bass possesses a tripartite Vtg system (VtgAa, VtgAb, and VtgC) in which all three forms of Vtg make a substantial contribution to the yolk. The production of Vtg-derived YPs is generally similar to that described for other perciforms. However, novel amino-terminal labeling of oocyte YPs prior to MS/MS identified multiple alternative sites for cleavage of these proteins from their parent Vtg, revealing a YP mixture far more complex than reported previously. This approach also revealed that the major YP product of each form of striped bass Vtg, lipovitellin heavy chain (LvH), undergoes limited degradation to smaller polypeptides during oocyte maturation, unlike the case in marine fishes spawning buoyant eggs in which LvHAa undergoes extensive proteolysis to osmotically active free amino acids. These differences likely reflect the lesser need for hydration of pelagic eggs spawned in freshwater. The detailed characterization of Vtgs and their proteolytic fate(s) during oocyte growth and maturation establishes striped bass as a freshwater model for investigating teleost multiple Vtg systems. 相似文献
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通过组织切片、酶联免疫吸附及荧光定量PCR等方法,研究了尼罗罗非鱼(Oreochromis niloticus)卵巢发育过程中性类固醇激素(雌二醇E2、孕酮P)、卵黄蛋白原(Vtg)含量及Vtg mRNA相对表达水平变化规律。结果表明,尼罗罗非鱼性腺指数(GSI)与卵巢成熟发育间呈同步性变化,Ⅴ期达到峰值。血清中E2含量自卵巢Ⅱ期开始显著升高,Ⅳ期达到峰值,Ⅴ期后显著下降;血清P含量自卵巢Ⅱ期开始不断升高,Ⅴ期达到峰值,Ⅵ期显著下降;E2、P分别在卵巢成熟发育前期与后期发挥作用。肝中Vtg含量先升、后降,Ⅳ期达峰值;血清、卵巢中Vtg含量均自Ⅱ期开始增加,Ⅴ期达峰值,Ⅵ期显著降低;不同组织中Vtg含量变化与卵巢成熟发育间存在密切关联。肝中Vtg mRNA表达水平在Ⅲ期达到峰值,Ⅳ期后持续下调;卵巢中Vtg mRNA表达水平相对较低,Ⅴ期达到峰值;初步推测肝、卵巢同是尼罗罗非鱼Vtg合成部位,肝是Vtg合成的主要器官,在卵黄积累阶段最为活跃,而卵巢Vtg合成水平相对较低。 相似文献
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Pituitary gonadotropin follicle-stimulating hormone (FSH) was identified in fish two decades ago, but its functional importance
in fish reproduction remains poorly defined, especially in non-salmonid species. This gap in our knowledge is partially due
to the lack of the hormone in pure form in most of the species studied. We describe here the production of two different forms
of biologically active recombinant zebrafish FSH (zfFSH and zfFSHHIS) using methylotrophic yeast, Pichia pastoris, as the bioreactor. One form (zfFSH) was produced as the molecule closer to the native form, with the two subunits (Cga and
Fshb) expressed separately under different promoters. The other form (zfFSHHIS) was produced as a single polypeptide, with the cDNAs for the two subunits joined to form a fusion gene that contained a
6X His tag as part of the linker between the two subunits. The culture conditions were optimized for pH and incubation time
for maximal production of the proteins. Using a zebrafish FSH receptor (Fshr)-based reporter gene assay, we tested and compared
the biological activities of the two forms of recombinant zebrafish FSH. Our results provide useful information for the future
production of recombinant gonadotropins in other fish species. 相似文献
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为了解岱衢洋海域鱼卵、仔稚鱼的群落结构及其与环境因子的关系,本团队分别在2010年5月—2012年2月春、夏、秋和冬季,利用大型浮游生物网在此海域开展了8个航次调查。共采集到鱼卵1 042粒,仔稚鱼2 055尾,隶属于10目19科37种。鱼卵优势种为扁舵鲣和小带鱼等;仔稚鱼的优势种为鰉、鳀和中华小公鱼等。调查海域鱼卵的平均密度是8.40粒/100 m3,仔稚鱼的平均密度是14.85尾/100 m3。单因素方差分析结果显示,2010年和2011年春、夏季4个季节之间丰富度指数(D)、均匀度指数(J′)和多样性指数(H′)均存在极显著差异。鱼卵、仔稚鱼资源密度分布不均匀,春、夏季较高,冬季最低,大体呈现南高北低的趋势。Pearson相关性结果分析得出,与鱼卵、仔稚鱼关系最密切的环境因子为温度、盐度和海水悬浮物。研究表明,春、夏季是岱衢洋海域鱼类产卵的重要时期,该海域是鳀、扁舵鲣、中华小公鱼等中上层鱼类的重要产卵场和育幼场。 相似文献