Effects of dietary estradiol-17β on feminization,growth and body composition in the Japanese eel (Anguilla japonica)

• 1.1.Juvenile Japanese eels (Anguilla japonica) were fed on a diet supplemented with estradiol-17β (E₂) at doses of 25, 50 and 75 mg/kg. The effects on growth, sex distribution and body composition were investigated in two groups of gonadally undifferentiated stages (early and later juvenile stages).
• 2.2.Feminization (95–100%) was observed in all E₂-treated groups.
• 3.3.The growth rate of fish treated with 25 and 50 mg/kg E₂ diet at the early juvenile stage was significantly increased.
• 4.4.The amount of protein in muscle decreased and that of fat increased in the E₂-treated groups except in the early juvenile stage fed with 25 mg/kg E₂.

     

How have spawning ground investigations of the Japanese eel Anguilla japonica contributed to the stock enhancement?

After Schmidt’s discovery of the spawning area of the Atlantic eels Anguilla anguilla and A. rostrata, the search for the Japanese eel A. japonica began in the Pacific Ocean. In 1991, the spawning area of the Japanese eel was determined to be the western North Pacific. Because of enthusiastic research, eggs and maturing eels have been collected in the Japanese eel. These findings are the first for one of the 19 freshwater eels. The population sizes of the Japanese and Atlantic eels are linearly decreasing. Thus, these eel population sizes are considered outside of safe biological limits, and the current fisheries are not sustainable. Artificial propagation has not yet succeeded for the freshwater eels. Stock assessment and management of the European eel have received increasing attention; however, such assessments and management of the Japanese eel have not yet been seriously considered. This paper is an overview of the results of intensive spawning ground investigations of the Japanese eel and describes how the outcomes of these studies have contributed not only to biological interests but also to stock enhancement. During the past 20 years of expeditions, noticeable findings have only been collected for wild eggs and mature adult specimens in spite of the expenditure of large research grants and the large amounts of time invested. The outcomes throughout an expedition do not necessarily contribute to the development and improvement of artificial breeding techniques and stock enhancement. Thus, eel research should be more focused on the studies related to eel stock management.     

Silvering and gill “mitochondria-rich” cells in the eel,Anguilla anguilla

Gills of typical yellow and silver ells, Anguilla anguilla L., were examined by light and electron microscopy. In both eel types, mitochondria-rich cells were located in the epithelium covering the primary lamellae and consisted ofchloride cells and accessory cells. As compared to yellow eels, the primary gill epithelium of silver eels was thicker and contained larger and more numerous chloride cells with enlarged mitochondria. The accessory cells also increased in number but did not show significant modifications in their size or ultrastructural features. These observations indicate that, as far as mitochondria-rich cells are concerned, the silvering process in eels would be equivalent to smoltification in salmonids. It corresponds to a preparation for seawater life and is probably controlled by hormonal factors.     

A morphometric analysis of the subcellular distribution of LHβ and FSHβ in secretory granules in the pituitary gonadotrophs of the frog (Rana japonica)

Immunohistochemical localization of lutropin (LH) and follitropin (FSH) in the pituitary gland of the frog Rana japonica was studied by the peroxidase-anti-peroxidase method and the two-face, double-labeling method with different-sized gold particles at the light-and electron-microscopic levels, respectively, using monoclonal antibodies against bullfrog LH and FSH. Light-microscopic immunohistochemistry indicated that approximately 66.0% of all the gonadotrophs in the pituitary contained both LH and FSH, whereas 33.4% of gonadotrophs contained only LH, and 0.6% contained only FSH. The staining intensity of LH and FSH varied from cell to cell. The gonadotrophs were classified into four types (Types I–IV) in terms of their ultrastructural and immunolabeling characteristics. Moreover, several secretory granule types were recognized according to differences in their shape and electron density. In all the cell types, both LH and FSH were often seen in the same secretory granules, but the proportion of granules bearing both hormones ranged from 5.5% in Type I to 32.7% in Type IV. Most secretory granules in Types I and II were immunolabeled with LH alone, whereas a small number of granules were immunolabeled with FSH alone. More immunolabeled FSH granules were present in Types III and IV than in Types I and II.     

Abundance and individual size of American eel (Anguilla rostrata) in the St. Lawrence River over the past four decades

Changes in abundance indicators and the mean size of American eel (Anguilla rostrata LeSueur) in the lower St. Lawrence River (SLR) over the past 40 years were assessed to determine the chronology of the species decline and the possible effect of hydroclimatic factors on that trend. Daily catches at commercial weirs and at an experimental fishery were used to model the long-term trend in silver eel availability. Eel landings started declining in the mid-1980s; whereas the modelled catch-per-unit-effort (CPUE) index shows a monotonic declining trend since the late 1960s. Landings and CPUEs were not strongly correlated, and landings cannot be a valid proxy for silver eel abundance in the SLR. Variations in summer river discharge contributed to interannual fluctuations in CPUE by affecting eel availability to the fishing gears. No relationship was found between CPUE and the North Atlantic Oscillation index of climatic variability. Correlations between hydroclimatic variables lagged by 1–20 years and CPUE indices revealed no measurable effect of riverine hydroclimatic variability on eel recruitment. The average size of silver eels increased by 30% between 1997 and 2007, as a result of poor recruitment during the mid-1980s. Since the onset of the silver eel decline preceded the major decline in recruitment in the mid-1980s, we suggest that the decline in spawning stock size was not due to poor recruitment but rather to large-scale mortality factors associated with high exploitation in upstream Lake Ontario and to construction of hydropower dams in the late 1950s.     

Is the continental life of the European eel Anguilla anguilla affected by the parasitic invader Anguillicoloides crassus?

Quantifying the fitness cost that parasites impose on wild hosts is a challenging task, because the epidemiological history of field-sampled hosts is often unknown. In this study, we used an internal marker of the parasite pressure on individual hosts to evaluate the costs of parasitism with respect to host body condition, size increase and reproductive potential of field-collected animals for which we also determined individual age. In our investigated system, the European eel Anguilla anguilla and the parasitic invader Anguillicoloides crassus, high virulence and severe impacts are expected because the host lacks an adaptive immune response. We demonstrated a nonlinear relationship between the severity of damage to the affected organ (i.e. the swimbladder, our internal marker) and parasite abundance and biomass, thus showing that the use of classical epidemiological parameters was not relevant here. Surprisingly, we found that the most severely affected eels (with damaged swimbladder) had greater body length and mass (+11% and +41%, respectively), than unaffected eels of same age. We discuss mechanisms that could explain this finding and other counterintuitive results in this host–parasite system, and highlight the likely importance of host panmixia in generating great inter-individual variability in growth potential and infection risk. Under that scenario, the most active foragers would not only have the greatest size increase, but also the highest probability of becoming repeatedly infected—via trophic parasite transmission—during their continental life.     

Expression of K-Cl cotransporters in the α-cells of rat endocrine pancreas

The expression of K⁺-Cl⁻ cotransporters (KCC) was examined in pancreatic islet cells. mRNA for KCC1, KCC3a, KCC3b and KCC4 were identified by RT-PCR in islets isolated from rat pancreas. In immunocytochemical studies, an antibody specific for KCC1 and KCC4 revealed the expression of KCC protein in α-cells, but not pancreatic β-cells nor δ-cells. A second antibody which does not discriminate among KCC isoforms identified KCC expression in both α-cell and β-cells. Exposure of isolated α-cells to hypotonic solutions caused cell swelling was followed by a regulatory volume decrease (RVD). The RVD was blocked by 10 μM [dihydroindenyl-oxy] alkanoic acid (DIOA; a KCC inhibitor). DIOA was without effect on the RVD in β-cells. NEM (0.2 mM), a KCC activator, caused a significant decrease of α-cell volume, which was completely inhibited by DIOA. By contrast, NEM had no effects on β-cell volume. In conclusion, KCCs are expressed in pancreatic α-cells and β-cells. However, they make a significant contribution to volume homeostasis only in α-cells.     

Roles of yeast eIF2α and eIF2β subunits in the binding of the initiator methionyl-tRNA

Heterotrimeric eukaryotic/archaeal translation initiation factor 2 (e/aIF2) binds initiator methionyl-tRNA and plays a key role in the selection of the start codon on messenger RNA. tRNA binding was extensively studied in the archaeal system. The γ subunit is able to bind tRNA, but the α subunit is required to reach high affinity whereas the β subunit has only a minor role. In Saccharomyces cerevisiae however, the available data suggest an opposite scenario with β having the most important contribution to tRNA-binding affinity. In order to overcome difficulties with purification of the yeast eIF2γ subunit, we designed chimeric eIF2 by assembling yeast α and β subunits to archaeal γ subunit. We show that the β subunit of yeast has indeed an important role, with the eukaryote-specific N- and C-terminal domains being necessary to obtain full tRNA-binding affinity. The α subunit apparently has a modest contribution. However, the positive effect of α on tRNA binding can be progressively increased upon shortening the acidic C-terminal extension. These results, together with small angle X-ray scattering experiments, support the idea that in yeast eIF2, the tRNA molecule is bound by the α subunit in a manner similar to that observed in the archaeal aIF2–GDPNP–tRNA complex.     

An autoradiographic study of α1-adrenergic receptors in the brain of the Japanese quail (Coturnix coturnix japonica)

Summary The neuroanatomical distribution of ₁-adrenergic receptors was studied in Japanese quail by quantitative in vitro autoradiography using the specific antagonist [³H]prazosin as the ligand. The presence of saturable (B_max <200 fmol/mg protein) high affinity (K_d < 0.12 nM) binding sites was detected by saturation analysis. High concentrations of [³H]prazosin binding sites were detected in the archistriatum/pars ventralis, the hippocampus, the cortex piriformis, the area corticoidea dorsolateralis, the dorsal thalamus, and the nucleus praetectalis. Lower concentrations were seen in the intercollicular nucleus, the lateral septum, and the posterior and tuberal hypothalamus. Very little binding was seen in the preoptic and anterior hypothalamic areas. The relatively high number of binding sites identified in the telencephalic structures agrees well with previous mammalian studies. This is in contrast with the pattern in the anterior hypothalamus where, in mammals, a number of nuclei have been reported to contain a high receptor density.     

Too short to spawn? Implications of small body size and swimming distance on successful migration and maturation of the European eel Anguilla anguilla

Individual net fat reserves after migration and reproductive investments were calculated for migrating female silver eels Anguilla anguilla (n = 387) collected in the outlet region of the Baltic Sea during the autumn run. It is estimated that 20·4% of the A. anguilla had completely exhausted all initial fat reserves and that 45·0% of A. anguilla were within 90% of complete energy depletion after migration and reproduction. This study concludes that a combination of body size and distance (6900 km) to the spawning area in the Sargasso Sea explains the results. An increase in the costs of migration due to heavy infection with Anguillicoloides crassus was also evaluated in an additional scenario with results showing that 26·4% of the A. anguilla had completely depleted all fat reserves. It is hypothesized that a large proportion of female silver A. anguilla from the Baltic Sea catchment area will have inadequate or suboptimal reserves for successful migration and reproduction.     

Intracellular fate of fibrinogen Bβ chain expressed in COS cells

Full-length fibrinogen Bβ cDNA was subcloned into an expression vector, pBC12BI, and transfected into COS cells. Bβ chain expression was measured by pulse-labelling cells with l-[³⁵S]methionine, immunoprecipitating the Bβ chain with antibody to fibrinogen and separating the nascent radioactive protein by sodium dodecyl sulfate-pholyacrylamine gel electrophoresis (SDS-PAGE). Bβ chain was expressed in transfected COS cells but was not secreted into the medium. Treatment with endoglycosidase H showed that non-secreted Bβ chain contains mannose-rich carbohydrates rather than the complex form of carbohydrate which occurs in plasma fibrinogen and indicates that Bβ chain is not transported to the Golgi apparatus. In transfected COS cells, anitibody to fibrinogen co-immunoprecipitated Bβ chain and 78 kDa immunoglobulin-binding protein (BiP) and antibody to BiP immunoprecipitated BiP and nascent Bβ chains. Non-secreted Bβ chain was degraded intracellularly with a half-life of 5 h by enzymes which were not affected by incubating transfected cells with NH₄Cl, which indicates a non-lysosomal pathway of degradation. These studies indicate that Bβ chain by itself does not contain the signal for fibrinogen secretion and that non-secreted Bβ chain is associated with BiP and degraded in the rough endoplasmic reticulum.     

Release of salmon gonadotropin and 17α-hydroxyprogesterone from lipophilized gelatin emulsion at 10, 20 and 30°C in the Japanese eel

Hormone releasing properties from an emulsion prepared with lipophilized gelatin (LG emulsion) were investigated on salmon gonadotropin (sGTH) as a peptide hormone and 17-hydroxyprogesterone (17P) as a steroid at 10, 20 and 30°C by monitoring plasma profiles of these hormones after administration in the Japanese eel. Release of these hormones from the LG emulsion were slow and not largely influenced by water temperature, whereas release from saline solution (sGTH) or cottonseed oil (17P) were rapid and increased with the elevation of temperature. © Rapid Science Ltd. 1998     

Lipid and fatty acid variations in muscle tissues of the ‘yellow’ stage of the European eel (Anguilla anguilla) during short-term adaptation to freshwater and seawater under food deprivation

Changes in body mass and water content (WC), lipids, and fatty acid (FA) composition of muscle were determined in European eels (Anguilla anguilla) in the ‘yellow’ stage (sexually immature). The animals were held in either freshwater (FW) or seawater (SW) and deprived of food for four weeks. During this period, body mass decreased steadily. This decrease was accompanied by reduced muscle WC and total lipid in both SW and FW. ‘Yellow’ eels appeared to use lipids in muscle to provide metabolic and osmoregulatory energy. However, they maintained relative constancy in the FA profiles, so that total FA, saturated, monounsaturated, and polyunsaturated FA, were similar between salinities during short-term starvation.     

Ageing sensitized by iPLA2β deficiency induces liver fibrosis and intestinal atrophy involving suppression of homeostatic genes and alteration of intestinal lipids and bile acids

Ageing is a major risk factor for various forms of liver and gastrointestinal (GI) disease and genetic background may contribute to the pathogenesis of these diseases. Group VIA phospholipase A2 or iPLA₂β is a homeostatic PLA₂ by playing a role in phospholipid metabolism and remodeling. Global iPLA₂β^−/− mice exhibit aged-dependent phenotypes with body weight loss and abnormalities in the bone and brain. We have previously reported the abnormalities in these mutant mice showing susceptibility for chemical-induced liver injury and colitis. We hypothesize that iPLA₂β deficiency may sensitize with ageing for an induction of GI injury. Male wild-type and iPLA₂β^−/− mice at 4 and 20–22 months of age were studied. Aged, but not young, iPLA₂β^−/− mice showed increased hepatic fibrosis and biliary ductular expansion as well as severe intestinal atrophy associated with increased apoptosis, pro-inflammation, disrupted tight junction, and reduced number of mucin-containing globlet cells. This damage was associated with decreased expression of intestinal endoplasmic stress XBP1 and its regulator HNF1α, FATP4, ACSL5, bile-acid transport genes as well as nuclear receptors LXRα and FXR. By LC/MS-MS profiling, iPLA₂β deficiency in aged mice caused an increase of intestinal arachidonate-containing phospholipids concomitant with a decrease in ceramides. By the suppression of intestinal FXR/FGF-15 signaling, hepatic bile-acid synthesis gene expression was increased leading to an elevation of secondary and hydrophobic bile acids in liver, bile, and intestine. In conclusions, ageing sensitized by iPLA₂β deficiency caused a decline of key intestinal homeostatic genes resulting in the development of GI disease in a gut-to-liver manner.