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1.
Flatfish or flounder moves one eye to change body proportion into vertebral asymmetry during metamorphosis, during which some
become sinistral while others dextral. However, the mechanism behinds the eye-position has not been well understood. In this
research, hybrids between Japanese flounder(♀) and stone flounder (♂) show mixed eye-location in both dextral type and sinistral
type, and thus become good samples for studying the eye-migration. mRNAs from pro-metamorphosis sinistral and dextral hybrids
larvae were screened with classical differential display RT-PCR (DD-RT-PCR) and representational difference analysis of cDNA
(cDNA-RDA); 30 and 47 putative fragments were isolated, respectively. The cDNA fragments of creatine kinase and trypsinogen
2 precursor genes isolated by cDNA-RDA exhibited eye-position expression patterns during metamorphosis. However, none of the
fragments was proved to be related to flatfishes’ eye-position specifically. Therefore, further studies and more sensitive
gene isolated methods are needed to solve the problems.
Supported by National Natural Science Foundation of China (No. 30600455) and the National Basic Research Program of China
(973 Program, No.2004CB117402) 相似文献
2.
YOU Feng WANG Wei XU Dongdong ZHU Xiangping NI Jing WU Zhihao XU Yongli WANG Xincheng ZHANG Peijun 《中国海洋湖沼学报》2009,27(2)
The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder Sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes,12 confirmed hybridization of the paternal and matemal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish. 相似文献
3.
Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus： karyotype, allozyme and RAPD analyses 总被引：2，自引：0，他引：2
The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder sperm. Sinistral and dextral are two types
of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species.
Of the 22 loci examined from 12 allozymes, 12 confirmed hybridization of the paternal and maternal loci in hybrids and no
difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also
studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal
and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above
38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation
could inherit a little more genetic materials from paternal fish than that from maternal fish.
Supported by National Natural Science Foundation of China (No. 30571445), National High-Technology Research and Development
Program (863 Program, No. 2006AA10A404), and Project from the Ministry of Science and Technology of China (No. 2006DKA30470-017) 相似文献
4.
The symmetrical body of flatfish larvae changes dramatically into an asymmetrical form after metamorphosis. The molecular mechanisms responsible for this change are poorly understood. As an initial step to clarify these mechanisms, we used representational difference analysis of cDNA for the identification of genes active during metamorphosis in the Japanese flounder, Paralichthys olicaceus. One of the up-regulated genes was identified as creatine kinase muscle type 1 (CK-M1). Sequence analysis of CK-M1 revealed that it spanned 1 708 bp and encoded a protein of 382 amino acids. The overall amino acid sequence of the CK-M1 was highly conserved with those of other organisms. CK-M1 was expressed in adult fish tissues, including skeletal muscle, intestine and gill. Whole mount in-situ hybridization showed that the enhanced expression of CK-M1 expanded from the head to the whole body of larvae as metamorphosis progressed. Quantitative analysis revealed stage-specific high expression of CK-M1 during metamorphosis. The expression level of CK-M1 increased initially and peaked at metamorphosis, decreased afterward, and finally returned to the pre-metamorphosis level. This stage-specific expression pattern suggested strongly that CK-M1 was related to metamorphosis in the Japanese flounder. Its specific role in metamorphosis requires further study. 相似文献
5.
ZHUJie ZHANGXiumei GAOTianxiang 《中国海洋大学学报(英文版)》2005,4(2):145-151
Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigmcnt cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time. 相似文献
6.
遥感技术在西藏当雄地区活动断裂调查中的应用 总被引：1，自引：0，他引：1
以遥感数据、地震资料以及震源机制解为基础,对西藏当雄地区亚东-谷露断裂以及嘉黎断裂进行遥感解译和对比分析。研究表明,亚东-谷露断裂北段地貌特征为正断拉张及右旋走滑综合作用的结果;中段存在拉张运动与左旋走滑运动,并发育地堑,冲沟位错显示该处具有6～66 m的水平位移量,其中1411年当雄南8级地震在该处产生6～12 m的水平位移;南段次级断层充分发育,北端表现为右旋运动,往南开始表现出左旋运动特征,同时地堑充分发育。结合震源机制解反演结果,推测走滑断层控制该区地壳浅部（8 km以上）以走滑型为主的地震事件的发生,而该处的地堑形成于地壳深部（8 km以下）的正断型应力机制,并控制地壳深部正断型地震事件的发生。嘉黎断裂整体断层隐伏,西段存在右旋运动,往东南则开始出现左旋运动,该处地震事件和应力机制均为走滑型。 相似文献
7.
We isolated a strain of lymphocystis disease virus (LCDV) from Japanese flounder (Paralichthys olivaceus) cultured in northern China. Based on published sequences of major capsid protein (MCP) gene of LCDV-cn (GenBank: AF126405),
we designed two primer sets P1/P2 and P3/P4. We then used one-step or nested PCR and in-situ hybridization (ISH) to detect
LCDV and identify the target tissues or cells in infected Japanese flounder. The PCR products were positive in purified viral
supernatant, skin nodules, gut, gill, kidney, spleen, stomach, heart, and liver of Japanese flounder. We compared the DNA
sequence with 14 MCP nucleotide sequences from GenBank, including Megalocytivirus (OFIV and RSIV), Iridovirus (CzIV and WIV), Ranavirus (TFV and FV3), and Lymphocystivirus (8 LCDV). Based on the alignment, we confirmed the PCR product was from Lymphocystivirus (GenBank accession number DQ279090 (LCDV-HD)). Using ISH, we noted the presence of LCDV in the skin nodules, gut, gill, spleen,
stomach, and heart of spontaneously infected Japanese flounders. We successfully amplified LCDV fragments from Schlegel’s
black rockfish (Sebastes schlegeli Higendorf), redwing sea robin (Lepidotrigla microptera Günther) and turbot (Scophthalmus maximus) using the one-step and nested PCR, suggesting the target genes can be widely detected in fish using this method. 相似文献
8.
Insulin-like growth factors (IGFs) are key regulators of development and growth. Here, we characterized the igf2 gene from olive flounder (Paralichthys olivaceus) and determined its temporal and spatial expression. We set up an in-vitro protein expression system in eukaryotic human embryonic kidney (HEK293T) cells and explored its effects on cell proliferation. The flounder igf2 cDNA contained a 648-bp open reading frame (ORF) encoding a protein of 215 amino acids (aa), which spanned the complete signal peptide (47 aa), mature peptide (70 aa), and E domain (98 aa). In adult flounder, igf2 mRNA was detected in all selected tissues. In early development, igf2 mRNA was detected throughout development from unfertilized eggs to hatching-stage embryos. In-situ hybridization analysis indicated that igf2 mRNA was specially expressed in the brain region, floor plate, hypochord, otic vesicle, and pectoral fin during embryogenesis. Western blotting analysis indicated that the soluble recombinant flounder IGF2 protein was successfully produced through eukaryotic expression in HEK293T cells. In addition, the recombinant IGF2 protein significantly promoted the proliferation of human cervical carcinoma (HeLa) and HEK293T cells. These results provide new information about the structural and functional conservation, expression patterns, and biological activity of the igf2 in teleosts.
相似文献9.
ZHU Jie ZHANG Xiumei and GAO Tianxiang Fisheries College Ocean University of China Qingdao P.R.China 《中国海洋大学学报(英文版)》2005,4(2)
Histological development of Japanese flounder Paralichthys olivaceus larval skin and ultrastructural difference of skin between reared normal and malpigmented Japanese flounder were studied with light microscopy (LM) and transmission electron microscopy (TEM). The results show that the skin develops slowly before the metamorphosis, while at the onset of metamorphosis, the skin develops quickly and becomes complete in structure till about 50d after being hatched. Ultrastructural observation on the normal and malpigmented skins shows that the iridophore and melanophore are adjacent to each other. Profile and structure of the two kinds of pigment cells are more complete in the skin of normal ocular side than in the skin of pigmented blind side. The ultrastructure of typical chloride cell was observed in the skin of Japanese flounder larvae for the first time. 相似文献
10.
LI Yong SU De-chen ZHOU Rong-jun LI Hai-bing Alexander L.DENSMORE YAN Liang YAN Zhao-kun 《山地科学学报》2013,10(1):29-42
Longmen Mountain located at the boundary between the Sichuan Basin and Tibetan Plateau,representing the steepest gradient of any edges of the plateau.Three endmember models of uplift process and mechanism have been proposed,including crustal thickening,crustal flow,and crustal isostatic rebound.Here we use coeval sedimentary sequences in the foreland basin to restraint uplift process and mechanism in the Longmen Mountain.The more than 10,000 m thick Late TriassicQuaternary strata filled in this foreland basin and can be divided into six megasequences that are distinguished as two distinct types.The first type is the wedge-shaped megasequences which are sedimentary response of strong active thrust loading events,characterized by a high rate of subsidence and sediment accumulation,coarsening-upward succession and a dual-sourced sediment supply.This type includes Late Triassic,Late Jurassic to Early Cretaceous and Late Cretaceous to Paleogene megasequences.The second type is the tabular megasequences,characterized by the low rate of subsidence and sediment accumulation,finingupward succession,and a single-sourced sediment supply,which is sedimentary response of isostatic rebound and erosion unloading.This type includes the Early to Middle Jurassic,Middle Cretaceous and Neogene to Quaternary megasequences.Basing on sedimentary,active tectonic,geomorphic evidence,we infer that the direction has been reversed from SSWdirected sinistral strike-slip to NNE-directed dextral strike-slip during 40-3.6 Ma,and since 3.6 Ma,the Longmen Mountain thrust belt belong to times of isostatic rebound and erosional unloading with NNEdirected dextral strike-slip.This suggests that crustal isostatic rebound is a primary driver for uplift and topography of the present Longmen Mountain.The Wenchuan(Ms8.0) earthquake,which ruptured a large thrust fault with NNE-directed dextral strikeslip along the range front,is an active manifestation of this crustal isostatic rebound process with dextral strike-slipping and shortening.This process may be the cause for the Wenchuan Earthquake and the apparent paradox of high relief,little shortening,the relative dearth of historical seismicity in the region. 相似文献
11.
Neuropeptide Y(NPY) is a 36-amino acid peptide of the neuropeptide Y family that plays key roles in the regulation of food intake. In this study,we focused on NPY m RNA expression changes around feeding time and during food deprivation in olive flounder. The olive flounder NPY m RNA levels were analyzed in different tissues and a high level of expression was detected in the brain. We also demonstrated a correlation between NPY expression levels in the brain and feeding schedule. NPY expression levels in olive flounder maintained on a daily scheduled feeding regimen increased shortly before feeding and decreased after the scheduled feeding time. Compared with the-1 h group before feeding,NPY expression in the 3 h group after feeding decreased significantly( P <0.05). Food deprivation led to an 81.7% decrease in NPY m RNA levels in the 24 h fasted group(P <0.05) and a 91.7% decrease in the 48 h fasted group(P <0.05). Therefore,our study demonstrates that NPY expression is associated with food intake in olive flounder. This result reveals the function of NPY in regulating food intake and its potential importance in olive flounder aquaculture. 相似文献
12.
Extracellular products (ECP) produced byVibrio anguillarum strain M3 originally isolated from diseased flounder (Paralichthys olivaceus) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity
against azocasin was detected. Bacterium M2 showed highest growth and protease activity at 25°C. The protease present in ECP
showed maximal activity at pH 8 and 55°C; was completely inactivated by application of 80°C heat for 30 min; was completely
inhibited by EDTA and HgCl2, and was partially inhibited by PMSF, SDS, MnCl2 and iodoacetic acid; but not inhibited by CaCl2 and MgCl2. The ECP was toxic to flounder fish at LD50 values of 3.1 μg protein/g body weight. The addition of HgCl2 and application of heat at 50°C decreased the lethal toxicity of ECP. When heated at 100°C, ECP lethality to flounder was
completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including
necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerious lymphocytes
in the liver. These results showed that ECP protease was a lethal factor produced by the bacteriumV. anguillarum M3.
Contribution No. 4213 from the Institute of Oceanology, Chinese Academy of Sciences.
Funded by projects under the Major State Basic Research Development Program, Grant G1999012003. 相似文献
13.
To reveal the key factor in self-healing from LCDV (lymphocystis disease virus)-infec Japanese flounder (Paralichthys olivaceus), serum proteins from self-healing and sick Japanese floun were separated by two-dimensional electrophoresis to screen differentially expressed proteins. Prot spots demonstrating changes greater than two-fold in the expression level were digested and furt identified in capillary liquid chromatography tandem mass spectrometry (LC-MS/MS). Two immuni relevant proteins were thus identified as transferrin and the complement component C3 of Japan flounder. These findings suggest that the two proteins may play important roles in the self-healing lymphocystis in Japanese flounder. This is an important theoretical foundation to promote self-healing LCDV-infected Japanese flounder by improving their innate immunity. 相似文献
14.
15.
《中国海洋湖沼学报》2016,(5)
Ontogenetic changes in digestive enzyme activities and the amino acid(AA) profile of starry flounder,Platichthys stellatus,were investigated and limiting amino acids were estimated compared with the essential AA profile between larvae and live food to clarify starry flounder larval nutritional requirements.Larvae were collected at the egg stage and 0,2,4,7,12,17,24 days after hatching(DAH)for analysis.Larvae grew from 1.91 mm at hatching to 12.13 mm at 24 DAH.Trypsin and chymotrypsin activities changed slightly by 4 DAH and then increased significantly 4 DAH.Pepsin activity increased sharply beginning 17 DAH.Lipase activity increased significantly 4 DAH and increased progressively with larval growth.Amylase activity was also detected in newly hatched larvae and increased 7 DAH followed by a gradual decrease.High free amino acid(FAA) content was detected in starry flounder eggs(110.72mg/g dry weight).Total FAA content dropped to 43.29 mg/g in 4-DAH larvae and then decreased gradually to 13.74 mg/g in 24-DAH larvae.Most FAAs(except lysine and methionine) decreased 50%in 4-DAH larvae compared with those in eggs and then decreased to the lowest values in 24-DAH larvae.Changes in the protein amino acid(PAA) profile were much milder than those observed for FAAs.Most PAAs increased gradually during larval development,except lysine and phenylalanine.The percentages of free threonine,valine,isoleucine,and leucine decreased until the end of the trial,whereas the protein forms of these four AAs followed the opposite trend.A comparison of the essential AA composition of live food(rotifers,Artemia nauplii,and Artemia metanauplii) and larvae suggested that methionine was potentially the first limiting AA.These results may help develop starry flounder larviculture methods by solving the AA imbalance in live food.Moreover,the increased digestive enzyme activities indicate the possibility of introducing artificial compound feed. 相似文献
16.
Histological study on the ontogeny of the lymphoid organs, kidney, thymus and spleen of Japanese flounder, Paralichthys olivaceus, from hatching to 40 d was carried out. The pronephric kidney duct appeared early in hatching although the primordial haemopoietic
stem cells were observed within a week after hatching. The spleen was first seen after 8d of hatching. The thymus appeared
after 15d, situated near the pronephric kidney. Small lymphoid cells appeared during the later phase of the post-larval stage
in the sequence of thymus, kidney and spleen. During the 40d of observations, there were no distinct inner or outer zones
in thymus and no red or white pulp in spleen. These results suggest that the nonspecific defense immune system plays a very
important role in the early larval stage of Japanese flounder. 相似文献
17.
MyoD,Myf5,and myogenin are myogenic regulatory factors that play important roles during myogenesis.It is thought that MyoD and Myf5 are required for myogenic determination,while myogenin is important for terminal differentiation and lineage maintenance.To better understand the function of myogenic regulatory factors in muscle development of flounder,an important economic fish in Asia,real-time quantitative RT-PCR was used to characterize the expression patterns of MyoD,Myf5, and myogenin at early stages of embryo development,and in different tissues of the adult flounder.The results show that,Myf5 is the first gene to be expressed during the early stages of flounder development, followed by MyoD and myogenin.The expressions of Myf5,MyoD,and myogenin at the early stages have a common characteristic:expression gradually increased to a peak level,and then gradually decreased to an extremely low level.In the adult flounder,the expression of the three genes in muscle is much higher than that in other tissues,indicating that they are important for muscle growth and maintenance of grown fish.During embryonic stages,the expression level of MyoD might serve an important role in the balance between muscle cell differentiation and proliferation.When the MyoD expression is over 30%of its highest level,the muscle cells enter the differentiation stage. 相似文献
18.
Broodstock of Verasper moseri (Jordan and Gilbert) aged 3–4 years old were selected, and reinforced cultivation was conducted to promote maturation under
controlled water temperature and photoperiod conditions. Fertilized eggs were obtained by artificial fertilization, and the
development of embryos, larvae and juveniles was observed continuously. The results showed that the fertilized eggs of V. moseri were spherical, with transparent yolk and homogeneous bioplasm, and had no oil globule inside. The average diameter of the
eggs was 1.77±0.02 mm. The eggs of V. moseri were buoyant in water with salinity above 35. The cleavage type was typical discoidal. Young pigment cells appeared when
olfactory plates began to form. Hatching occurred at 187 h after fertilization at a water temperature of 8.5°C. The newly
hatched larvae, floating on the water surface, were transparent with an average total length of 4.69±0.15 mm. During the cultivation
period, when the water temperature was raised from 9 to 14.5°C, 4-day old larvae showed more melanophores on the body surface,
making the larvae gray in color. The pectoral fins began to develop, which enabled the larvae to swim horizontally and in
a lively manner. On days 7–8, the digestive duct formed. The yolk sac was small and black. The yolk sac was absorbed on day
11. Larvae took food actively, and body length and body height clearly increased. The rudiments of dorsal and anal fin pterygiophores
were discernible and caudal fin ray elements formed on day 19. On day 24, the larval notochord flexed upwards, and the rays
of unpaired fins began to differentiate. Pigment cells converged on the dorsal and anal fin rays, and the mastoid teeth on
the mandible appeared. On day 29, the left eyes of juveniles began to move upwards. Depigmentation began in some juveniles
and they became sandy brown in color on day 37. Most juveniles began to settle on the bottom of the tank. The left eyes of
juveniles migrated completely to the right side on day 50, when the average body length attained 2.5±0.18 cm, and juveniles
accomplished metamorphosis to young. The embryonic and larval characters of several flounder species are compared. 相似文献
19.
To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against
human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin
(PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder,Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like
factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes
and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation
assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2,
(rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface
of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected
with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%.
This work was supported by National “973” Project G1999012003, G19999012006. 相似文献
20.
宏、微观构造特征研究表明 ,红柳河地区造山期韧性变形大致可分两个阶段。早期形成南倾的面理、东倾的拉伸线理、鞘褶皱 ,变形具右行逆冲剪切特点 ,有限应变分析表明具压扁变形特征。晚期变形形成红柳河主干背斜、西倾的拉伸线理及红柳河—前进工区左行逆冲走滑剪切带。根据变形岩石和未变形岩石年龄确定变形期为加里东晚期—海西早期 ,变质变形特征显示变形层次属中浅构造层次 相似文献