Myotomal myogenesis of axial muscle in the sturgeon Acipenser baeri (Chondrostei, Acipenseriformes)

Compared to teleost fishes, a unique character of the myogenesis of the plesiomorphic A. baeri is the fusion of myoblasts derived from the somite, leading to the formation of multinucleate muscle lamellae. Then, the lamellae are converted into cylindrical muscle fibres. The mechanism of transformation of lamellae into fibres is still debatable. Early embryonic muscle growth is mainly due to the hypertrophy of somite-cell derived stock. After hatching, hypertrophic growth occurs parallel to hyperplastic growth. Proliferatively active mesenchymal cells, which migrate from the intermyotomal space into the myotomes, participate in both processes.     

The estrogenic activity of certain phytoestrogens in the Siberian sturgeon Acipenser baeri

Various phytoestrogens such as formononetin, daidzein, genistein and equol were synthesized. Their purity was assessed by various analytical techniques including melting point determination, thin-layer chromatography (TLC), infra-red spectra (i.r. spectra), nuclear magnetic resonance (1H- and 13C-NMR) and gas chromatography coupled with mass spectrometry (GC-MS). The estrogenic activity of these compounds, as well as biochanin A and coumestrol, was biologically tested by the induction of vitellogenin secretion in yearling sturgeon and compared to the activity of estradiol-17 beta. Pure daidzein, biochanin A, genistein, equol and coumestrol all had estrogenic activity as assessed by their induction of hepatic synthesis of vitellogenin when administrated intraperitoneally to yearling Siberian sturgeon. Coumestrol seemed to be the most potent compound, inducing the most vitellogenin secretion with the lowest dose administered. Formononetin was inactive when administered by the intraperitoneal route. All the phytoestrogens tested were considerably less potent than estradiol-17 beta.     

Calbindin D28k and calretinin in oxytocin and vasopressin neurons of the rat supraoptic nucleus

The aim of the present study was to examine quantitatively whether two calcium-binding proteins, calbindin D28k and calretinin, are localized in oxytocin and vasopressin neurons of the supraoptic nucleus of the male rat. We used a triple-labeling immunofluorescence method with a confocal laser scanning microscope. Of the oxytocin-labeled cells, 70% were stained for both calbindin D28k and calretinin, 15% were stained for only calbindin D28k, 13% were stained for only calretinin, and 2% were stained for neither protein. Of the vasopressin-labeled cells, 73% were stained for neither calbindin D28k nor calretinin, 21% were stained for only calbindin D28k, 4% were stained for only calretinin, and 2% were stained for both proteins. Calbindin D28k and calretinin have been shown previously to contribute to calcium homeostasis by buffering [Ca2+]i. Therefore, these findings suggest that most of the oxytocin neurons may have a higher Ca(2+)-buffering capacity than most of the vasopressin neurons.     

Characterization of xenobiotic metabolizing enzymes in sturgeon (Acipenser baeri)

1. Cytochrome P-450, NADPH-cytochrome c reductase, benzo(a)-pyrene hydroxylase (AHH), 7-ethoxycoumarin-O-deethylase (7-ECOD), epoxide hydrolase (EH), UDP-glucuronyltransferase (UDPGT) and glutathione S-transferase (GSHST) activities in sturgeon (Acipenser baeri) have been measured and partially characterized. 2. Cytochrome P-450-dependent monoxygenase (MO), EH, and conjugation reactions were detected in liver and to a lesser extent in kidney and gills. 3. Hepatic enzyme activities in the sturgeon were equally high or higher than in rainbow trout liver, with the exception of UDPGT whose activity was 14% of that in trout liver. 4. The MO and EH activities displayed the expected pH maxima of 7.5, whereas transferases were relatively independent of the pH in the 6.5-7.5 range. 5. The temperature optima for MO and EH were close to those reported in other fish species, whereas for conjugation reactions the temperature optima were 45 and 60 degrees C for GSHST and UDPGT respectively.     

Morphology of calretinin and tyrosine hydroxylase-immunoreactive neurons in the pig retina

The morphology of calretinin- and tyrosine hydroxylase-immunoreactive (IR) neurons in adult pig retina was studied. These neurons were identified using antibody immunocytochemistry. Calretinin immunoreactivity was found in numerous cell bodies in the ganglion cell layer. Large ganglion cells, however, were not labeled. In the inner nuclear layer, the regular distribution of calretinin-IR neurons, the inner marginal location of their cell bodies in the inner nuclear layer, and the distinctive bilaminar morphologies of their dendritic arbors in the inner plexiform layer suggested that these calretinin-IR cells were AII amacrine cells. Calretinin immunoreactivity was observed in both A-and B-type horizontal cells. Neurons in the photoreceptor cell layer were not labeled by this antibody. The great majority of tyrosine hydroxylase-IR neurons were located at the innermost border of the inner nuclear layer (conventional amacrines). The processes were monostratified and ran laterally within layer 1 of the inner plexiform layer. Some of the tyrosine hydroxylase-IR neurons were located in the ganglion cell layer (displaced amacrines). The processes of displaced tyrosine hydroxylase-IR amacrine cells were also located within layer 1 of the inner plexiform layer. Some processes of a few neurons were located in the outer plexiform layer. A very low density of neurons had additional bands of tyrosine hydroxylase-IR processes in the middle and deep layers of the inner plexiform layer. The processes of tyrosine hydroxylase-IR neurons extended radially over a wide area and formed large, moderately branched dendritic fields. These processes occasionally had varicosities and formed "dendritic rings". These results indicate that calretinin- and tyrosine hydroxylase-IR neurons represent specific neuronal cell types in the pig retina.     

Morphology, chemical contents and physiology of chondrostean fish sperm: a comparative study between Siberian sturgeon (Acipenser baerii) and sterlet (Acipenser ruthenus)

The present study was carried out with two sturgeon species, Siberian sturgeon (Acipenser baerii) and sterlet (A. ruthenus) to compare spermatological parameters to better understand inter‐species differences. Significant differences between morphometric features were observed such as acrosome length, acrosome width, head length, midpiece width and flagellar length, while midpiece length did not reveal such differences. The sterlet has a shorter spermatozoon than the Siberian sturgeon. Ultrastructural parameters vary significantly in terms of length of the nucleus, diameter of the endonuclear canals (EC), size of posterolateral projections (PP) and diameter of flagellum. Mean values for density of spermatozoa in the semen, seminal plasma pH, osmolality (mOsmol kg^?1), along with Ca²⁺, Na⁺, K⁺, Cl^? ions concentrations (mm ) were determined to be 0.61 ± 0.37 × 10⁹, 8.16 ± 0.18, 77.20 ± 52.28, 0.24 ± 0.06, 31.39 ± 10.21, 3.51 ± 1.10, 14.00 ± 4.30 in A. baerii and 0.41 ± 0.32 × 10⁹, 8.13 ± 0.19, 50.74 ± 6.27, 0.16 ± 0.11, 20.11 ± 3.78, 1.26 ± 0.54, 6.11 ± 0.60 in A. ruthenus, respectively. Significant differences were observed in Na⁺, K⁺ and Cl^? concentrations in the seminal plasma as well as in sperm velocity. The percentage of motile spermatozoa did not show any significant difference between the two species. Comparing the results of this study with published literature data on sturgeon spermatozoa reveals that morphological and ultrastructural parameters of spermatozoa together with some parameters of the seminal fluid and spermatozoa velocity can be used in comparative spermatology to better understand inter‐species differences. The observed biochemical and physiological differences should be also considered for the development of methods for controlled reproduction and for sperm cryopreservation techniques.     

Immunohistochemical Distribution of Cytochrome P4501A in Larvae and Fingerlings of the Siberian Sturgeon, Acipenser baeri

In this paper we investigate by means of immunohistochemistry, the tissue distribution of constitutive cytochrome P4501A (CYP1A), from hatching until 30 days posthatching in developing Siberian sturgeon, Acipenser baeri. For this purpose, a polyclonal (BN-1) antiserum developed against a conservative sequence of piscine CYP1A and a monoclonal (C10-7) antiserum directed against cod CYP1A were used on paraffin-embedded samples. From hatching onwards, distinct CYP1A immunoreactivity was distinctly observed in the following tissues and cells: envelope of oil droplets, matrix and syncytium of the yolk-sac, sinusoids, biliary epithelial cells and hepatocytes. In the digestive tract, buccopharyngeal, oesophageal, gastric and intestinal epithelia, as well as the cytoplasm and brush border of enterocytes were CYP1A-positive. Interestingly, gastric glands and melanin-plug present within lumen of the digestive system were strongly immunoreactive. Kidney (epithelia of renal tubules), gills (pillar and endothelial cells), skin (epithelial cells), muscle fibres of heart and eye (retina) were positive. In brain, we observed a strong CYP1A staining in the developing telencephalon and especially in olfactory system, as well as in those nerve fibres running ventrally toward the posterior brain. A strong CYP1A staining was observed in vascular endothelia of all organs/tissues, especially in the liver. In general, the intensity of CYP1A immunostaining increased during larval development, suggesting besides its known metabolic function (endogenous and/or exogenous), a possible participation of this heme-protein in control of cell division, regulation of growth and differentiation.     

Calbindin D28k, parvalbumin, and calretinin immunoreactivity in the main and accessory olfactory bulbs of the gray short-tailed opossum, Monodelphis domestica

The vertebrate main and accessory olfactory bulbs (MOB and AOB) are the first synaptic sites in the olfactory pathways. The MOB is a cortical structure phylogenetically well conserved in its laminar structure and overall synaptic organization, while the AOB has significant species variation in size. In order to better understand signal processing in the two olfactory systems and the species differences, immunocytochemical staining and analysis were done of the neuronal expression patterns of the calcium-binding proteins calbindin D28k (CB), parvalbumin (PV), and calretinin (CR) in the MOB and AOB in a marsupial species, the gray short-tailed opossum, Monodelphis domestica. In the MOB, antibody to CB labeled periglomerular cells, superficial short axon cells / Van Gehuchten cells; antibody to PV labeled Van Gehuchten cells; and antibody to CR immunostained periglomerular cells, superficial short axon cells / Van Gehuchten cells, and granule cells. In the AOB, CB immunoreactivity was detected in periglomerular cells and a subpopulation of granule cells; antibody to PV labeled the superficial short axon cells / Van Gehuchten cells and granule cells; and antibody to CR labeled a small number of periglomerular cells, superficial short axon cells / Van Gehuchten cells, and granule cells. These results showed that the patterns of CB, PV, and CR expression differ in the opossum main and accessory olfactory bulbs and differ from that in other animal species. These varying patterns of neuronal immunostaining may be related to the different functions of the main and accessory olfactory bulbs and to the differing signal processing features.     

全人工繁殖西伯利亚鲟仔稚鱼发育的异速生长

西伯利亚鲟（Acipenser baeri）是中国主要的淡水养殖对象之一,目前已经实现了苗种的全人工繁殖,为了改善西伯利亚鲟苗种培育技术,采用Image-Pro Plus 5.1软件显微拍照并对仔稚鱼的可量性状进行测量和数据处理,研究了全人工繁殖西伯利亚鲟仔稚鱼（0～53日龄）发育的异速生长及器官优先发育在早期生存和环境适应性上的意义。结果表明：在9、28、37日龄时分别出现3个全长生长拐点。因此,全长的生长可以分为4个阶段,每个阶段的平均增长率分别为0.83、0.79、2.68和1.71 mm·d^-1,其中第3个阶段的生长率明显高于其他阶段;体质量的增长可以用Gaussion方程进行拟合,相关系数R²=0.99;全长（FL）与体质量(BW)的关系为BW=0.2×10^-5(FL)2.72,为负异速生长;在早期发育过程中,仔鱼的许多关键器官的大小均存在异速生长现象,眼径在3～4日龄时,最先达到生长拐点,这意味着眼部是优先发育的,使其在出膜后就能有效地躲避敌害;口宽在17～18日龄时出现拐点,此时随着感觉器官和各鳍的不断完善,主动摄食能力不断加强;胸鳍、背鳍、臀鳍的长度也分别在16～17、13～14、21～22日龄时出现生长拐点,标志着其游泳能力已比较完善,可以有效地躲避敌害和获得食物,为其早期的生存提供了保障。在育苗生产实践中,可以根据生长和发育的阶段性和优先性,适时地和针对性地创造必要的生长发育条件,保证其重要器官得到优先发育,提高早期的成活率。     

Effect of Sublethal Concentrations of Mercury, Cadmium,and Copper Salts on the Lysozyme Content in Fry ofthe Lena River Sturgeon Acipenser baeri

The paper presents data on a change in lysozyme content in tissues of spleen, liver and heart in fry of the Lena River sturgeon exposed to the presence of sublethal concentrations of Hg²⁺, Cd²⁺, and Cu²⁺ under conditions of chronic experiment. It has been shown that the lysozyme content in fish tissues varies and has a phasic character. The amplitude of fluctuations of this parameter depends on the moment of sampling, nature of the toxicant, and structural-functional organization of the studied organs.     

Microchromosomes in holocephalian,chondrostean and holostean fishes

Chondrostean and holostean fish of today are leftover relics: they share some characteristics with the venturesome crossopterygian fish, which launched the evolution of terrestrial vertebrates about 280 million years ago. The chromosome complements and DNA values of one chondrostean and two holostean species as well as one holocephalian species were studied. Their DNA values varied from 37% to 50% of that of mammals, and three of the species contained dot-like microchromosomes in their diploid complements. Their genome size and karyological characteristics are quite similar to those possessed by one group of reptiles and by avian species.In Duarte, this work was supported by a grant CA-05138 from the National Cancer Institute, U.S. Public Health Service, and in part by a grant FR 00433-01, Animal Care Grant, N.I.H. In Northwood, this project was supported by the British Empire Cancer Campaign.Fellow of the Institute for Advanced Learning of the City of Hope Medical Center.     

西伯利亚鲟和鲫鱼脑型一氧化氮合酶cDNA片段克隆及序列分析

实验根据脑型一氧化氮合酶氨基酸高度保守序列设计一对简并引物,采用RT-PCR方法扩增并克隆了西伯利亚鲟(Acipenser baeri)和鲫鱼(Carassius auratus)的脑型一氧化氮合酶cDNA片段。西伯利亚鲟cD-NA片段为356 bp,鲫鱼cDNA片段为377 bp。氨基酸序列同源分析发现,西伯利亚鲟与斑马鱼(Brachydaniorerio)的脑型一氧化氮合酶同源性为70%,鲫鱼与斑马鱼的同源性为97.6%。     

Effects of dietary bovine lactoferrin on growth, physiological performance, iron metabolism and non-specific immune responses of Siberian sturgeon Acipenser baeri

The present experiment was carried out to investigate the effects of different levels of dietary lactoferrin (LF) on growth performance, physiological status, iron absorption and innate immune response of juvenile Siberian sturgeon Acipenser baeri. Fish were fed with six different rations including 0, 100, 200, 400, 800 and 1600 mg LF kg(-1) diet for 8 weeks. At the end of the experiment, samples were collected for estimating the physiological and immunological parameters. Dietary LF did not change the fish growth performance, hematological parameters, serum proteins or hepatic enzymes. Moreover, stress indicators (plasma cortisol, glucose and lactate) were not affected by dietary LF. The iron absorption of fish was considerably affected by LF; thus, plasma iron in LF-treatments greatly declined and the total iron binding capacity (TIBC) significantly increased in fish fed with 800 mg LF kg(-1). In addition, the liver iron content markedly increased in some LF-treatments, but the variation of muscle iron concentration in treatments was insignificant. The amount of mucus secretion and serum bactericidal activity rose in fish fed on dietary LF, although other non-specific immune responses such as mucus bactericidal activity, serum and mucus lysozyme activity, serum peroxidase, serum natural hemolytic complement activity and serum IgM were not influenced by LF. This study revealed the ability of dietary LF to sequester iron, which is an essential nutrient required for the growth of bacteria. LF was also shown to improve some physiological and immunological parameters of Siberian sturgeon, to some extent.     

Effects of dietary vitamin C on tissue ascorbate and collagen status in sturgeon hybrids (Acipenser ruthenus L. x Acipenser baeri Brandt)

The aim of this study was to determine the vitamin C status of various tissues, and collagen concentration in cartilage, of a sterlet ( Acipenser ruthenus L.) x Siberian sturgeon ( Acipenser baerii Brandt) hybrid fed different dietary vitamin C rations. Growth, vitamin C status and collagen concentration were measured in groups fed diets supplemented with 150, 300, 600 mg.kg^-1 ascorbate-2-polyphosphate (APP); 100, 200, 400 mg.kg^-1 ascorbate-2-monophosphate (AMP), 1000 mg.kg^-1 L-ascorbic acid (AA) and with an almost ascorbate free diet (Total AA > 5 mg.kg^-1), as a control.
No significant differences in growth were observed, and no external symptoms of scurvy developed in the vitamin C-free group. No reduction in total vitamin C concentration was revealed in the tissues examined, as compared with initial concentrations. Ascorbate-2-phosphate esters were only found in kidney or hepatopancreas of fish fed with these vitamin C derivatives. Significantly (p<0.05) lower contents of collagen were observed in the control group after 8 weeks feeding. No significant differences were found among the groups after 16 weeks although the collagen concentration in the cartilage was slightly lower in the control group without vitamin C supplementation.     

Verification of meiotic gynogenesis in Siberian sturgeon (Acipenser baeri Brandt) using microsatellite DNA and cytogenetical markers

Diploid gynogenesis was induced in Siberian sturgeon Acipenser baeri using ultra violet (UV)-irradiated bester ( Huso huso × Acipenser ruthenus ) sperm. The higher survival rate of meiotic diploids was noted after 1350 ergs mm⁻² UV irradiation. A total of 80 meiotic diploids of known parentage from two different experimental treatments were screened using microsatellite DNA and cytogenetical analysis, and uniparental transmission in meiotic diploids was confirmed.     

Growth performance,antioxidant and immune status of Siberian sturgeon (Acipenser baeri) fed diets containing green tea extract and oxidized fish oil

The present study was conducted to determine the effects of dietary oxidized fish oil (OFO) and green tea extract (GTE) on growth performance, antioxidant status and immunological parameters in Siberian sturgeon (Acipenser baeri). Fish weighing about 108 g were randomly assigned to one of nine dietary treatments (control diet without OFO and GTE; diets containing only 25 or 50 mg/kg of GTE; diets containing 50 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE; diets containing 100 mg/kg of OFO combined with 0, 25 or 50 mg/kg of GTE) with three replicates of 15 fish stocked in fiberglass tanks using a completely randomized design. Growth performance indices, immunological parameters and antioxidant status of all treatment groups were determined using appropriate assays and compared. Results showed that GTE could enhance growth performance. Feeding GTE alone or combined with the OFO could decrease the hepatosomatic index (HSI) and viscerosomatic index (VSI). Serum lysozyme activity and antibody level were increased significantly by GTE alone or combined with OFO compared to the oxidized diets (p < 0.05). Conversely, lipid peroxidation products and antioxidant enzyme activity were significantly lower in GTE treatment groups and those that received combination of GTE and OFO compared to the oxidized diets. Low‐density lipoprotein (LDL) level in GTE groups was lower than other treatment groups (p < 0.05) however, in OFO treatment groups the LDL level in fish serum was significantly increased in comparison with the other treatment groups (p < 0.05). GTE and OFO alone or combined together significantly decreased the levels of triglyceride and cholesterol compared to the control group (p < 0.05). In conclusion, immunological and biochemical parameters were improved by inclusion of GTE into Siberian sturgeon rancid diets which can protect fish from the oxidative damage challenged by dietary oil oxidation.