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Cloning and expression of three aquaporin homologues from the European eel (Anguilla anguilla): effects of seawater acclimation and cortisol treatment on renal expression.

BACKGROUND INFORMATION: The European eel (Anguilla anguilla) is able to osmoregulate over a wide range of environmental salinities from FW (freshwater) to hyperconcentrated SW (seawater). Successful acclimation is associated with strict regulation of ion and water transport pathways within key osmoregulatory epithelia to enable animals to survive the dehydrating or oedematous conditions. These observations suggested that homologues of the AQP (aquaporin) water channel family were expressed in the eel and that these proteins may contribute to the water transport and osmoregulation in all euryhaline teleosts. RESULTS: Complementary DNAs encoding a homologue of the mammalian aquaglyceroporins (termed AQPe) and two homologues of mammalian aquaporin-1 [termed AQP1 and AQP1dup (aquaporin-1 duplicate)] were isolated from the European eel. Northern-blot analysis revealed (i) two AQP1 transcripts exhibiting a wide tissue distribution, (ii) a single AQP1dup mRNA transcript found in the kidney and the oesophagus, and (iii) a single AQPe mRNA detectable mainly in the kidney and the intestine. The relative expression of isoforms within the kidney was AQP1dup>AQPe>AQP1. SW acclimation significantly reduced the abundance of AQP1, AQP1dup and AQPe transcripts in the kidney of yellow eels by approx. 72, 66 and 34% respectively, whereas the expression levels in silver eels were independent of salinity and equivalent to those observed in yellow SW-acclimated fish. AQP1 protein expression was primarily located within the vascular endothelium in yellow eels and the epithelial apical brush border in some renal tubules in silver eels. Infusion of cortisol into FW eels had no effect on AQPe mRNA expression, but induced significant decreases in AQP1 and AQP1dup mRNA levels in the kidney of yellow eels. Cortisol infusion had no effect on the expression of any isoform in the silver eels. CONCLUSIONS: These results suggest that SW-acclimation or cortisol infusion induces a down-regulation of renal AQP expression in yellow eels. However, the lower levels of aquaporin expression found within the silver eel kidney were not further reduced by salinity transfer or steroid infusion. These differences in mRNA expression were accompanied by changes in the cellular distribution of the AQP1 protein between vascular endothelial and tubular epithelial cells.

Acclimatization↗

Dynamics and predicted decline of Anguillicola crassus infection in European eels, Anguilla anguilla, in Neusiedler See, Austria.

The eel population in Neusiedler See has been maintained by regular massive stocking since 1958. After the establishment of the National Park Neusiedler See-Seewinkel in 1993, eel stocking was prohibited and the population, together with the specific parasites of eels, was predicted to decline to extinction within 10 years. This investigation was undertaken to document the decline and extinction of the Anguillicola crassus population in eels. From 1994 to 2001, 720 eels were collected from two sites in the lake. Prevalence and abundance of A. crassus were lower in spring than in summer and autumn and larger eels harboured more parasites than smaller ones. Neither year of study nor sampling site were correlated with parasite infection levels. No significant trend in the population parameters of A. crassus was detected over the 8 years of the survey. This suggested that there had been no significant decline in the eel population. This suggestion was confirmed by investigations of the fishery, which also found evidence of regular illegal stocking. The stability of the A. crassus population over the past decade seems to reflect the lack of change in eel population density. No mass mortalities of eels occurred over the period despite the many similarities between Neusiedler See and Lake Balaton in Hungary. Differences in eel size, eel diet and the lack of large-scale insecticide use are discussed as possible explanations for the absence of eel mass mortalities in Neusiedler See.

Anguilla↗

Occurrence of Anguillicola crassus (Nematoda: Dracunculoidea) in Japanese eels Anguilla japonica from a river and an aquaculture unit in SW Taiwan.

The infection by swimbladder nematodes of the genus Anguillicola (Dracunculoidea: Anguillicolidae) was examined in 2 populations of the Japanese eel Anguilla japonica in SW Taiwan. Wild eels from the Kao-Ping river were compared with cultured eels from an adjacent aquaculture unit. Only the cosmopolitan species Anguillicola crassus was present. Among wild eels, prevalence of infection varied between 21 and 62%, and mean intensity between 1.7 and 2.7 for adult worms. Similar intensity values (1.3 to 2.8) were recorded for the larvae. In cultured eels, prevalence as well as mean intensities were higher. In the cultured hosts, mean larval intensities exceeded those of adult worms 2-fold, and maximum larval intensities were 4- to 5-fold higher than in eels from the river. In cultured eels, dead larvae were also more abundant than in wild eels. We conclude that infrapopulations of A. crassus in Japanese eels are regulated by the defense system of this host, intraspecific density-dependent regulation being less likely as the major regulatory mechanism. No influence of the parasite on eel condition was found in either wild or cultured eels, indicating a low or moderate pathogenic effect of A. crassus on this host. This study shows that A. crassus is moderately common in cultured and wild Japanese eels in Taiwan, where the parasite is endemic.

Air Sacs↗

Identification of eel ghrelin in plasma and stomach by radioimmunoassay and histochemistry.

The structure of ghrelin has been determined in the Japanese eel, Anguilla japonica. In this study, we identified immunoreactive ghrelin in extracts from plasma and stomach of the eel by radioimmunoassay (RIA) using an antiserum against octanoylated rat ghrelin [1-11]. Using the antiserum, we examined localization of ghrelin-immunopositive cells in the eel stomach. Detection of ghrelin mRNA-expressing cells was also attempted in the eel stomach using a cRNA probe specific for the eel ghrelin gene. Furthermore, we examined ghrelin expression patterns in plasma and stomach after transfer of freshwater (FW) eels to seawater (SW). Multiple types of immunoreactive ghrelin were detected using RIA. These were octanoylated eel ghrelin and other ghrelins that may have different fatty acid modifications, suggesting that this RIA can detect acylated ghrelin of eels as seen previously in the case of rat. Ghrelin-immunopositive cells were observed in the mucosal layer of the stomach, especially in the neck of the fundic gland. Ghrelin mRNA-expressing cells showed similar distribution and characteristics to the immunopositive cells. Plasma ghrelin levels in FW eels starved for one week before experimentation were approximately 40 fmol/ml. Plasma ghrelin levels in control-transferred FW eels did not change for 7 days, but significantly increased on day 14. Plasma ghrelin levels transiently increased fivefold 6h after SW transfer and then declined to the FW level by 24h after transfer. Ghrelin content and ghrelin mRNA levels in the stomach did not change after SW transfer, except for a transient decrease in ghrelin content seen 24h after transfer. The present results suggest that ghrelin may participate in osmoregulation in eels.

Acclimatization↗

Vibrio vulnificus biogroup 2: new biogroup pathogenic for eels.

Clinical and nonclinical isolates of the lactose-positive Vibrio vulnificus were compared with Vibrio strains isolated from lesions on eels (Anguilla japonica) cultured commercially in Japan. Strains were compared phenotypically and antigenically, for pathogenicity to mice and eels, and for genetic relatedness. The strains isolated from diseased eels differed phenotypically from the original species description of V. vulnificus in that they were negative for indole production, ornithine decarboxylase activity, growth at 42 degrees C, and acid production from mannitol and sorbitol. No relationship between the surface antigens of V. vulnificus strains from environmental and clinical sources and the strains from diseased eels was observed. Typical V. vulnificus strains and the eel isolates were pathogenic to mice; however, only those strains originally isolated from diseased eels were found to be pathogenic to eels. Results of DNA-DNA competition experiments revealed that there was greater than 90% relative reassociation between clinical and nonclinical V. vulnificus and strains from diseased eels. Based on the results of the DNA-DNA competition experiments, we conclude that the strains isolated from diseased eels were V. vulnificus; however, the differences in phenotypic characteristics and eel pathogenicity indicated that these strains represent a different biogroup. Therefore, we propose that strains phenotypically similar to the type strain of the species (ATCC 27562) be classified as V. vulnificus biogroup 1 and the strains phenotypically similar to those isolated from diseased eels be classified as V. vulnificus biogroup 2 represented by the reference strain ATCC 33148.

Animals↗

Angiotensin II modulates the activity of the Na+/K+ATPase in eel kidney.

We have previously shown that angiotensin II (Ang II) has a role at the level of the eel gill chloride cell regulating sodium balance, and therefore osmoregulation; the purpose of the present study was to extend these findings to another important osmoregulatory organ, the kidney. By catalytic histochemistry Na(+)/K(+)ATPase activity was found in both sea water (SW)- and freshwater (FW)-adapted eel kidney, particularly at the level of both proximal and distal tubules. Quantitation of tubular cell Na(+)/K(+)ATPase activity, by imaging, gave values in SW-adapted eels which were double those found in FW-adapted eels (Student's t-test: P<0.0001). This was due to a reduced number of positive tubules present in FW-adapted eels compared with SW-adapted eels. By conventional enzymatic assay, the Na(+)/K(+)ATPase activity in isolated tubular cells from SW-adapted eels showed values 1.85-fold higher those found in FW-adapted eels (Student's ttest: P<0.0001). Perfusion of kidney for 20 min with 100 nM Ang II provoked a significant increase (1.8-fold) in Na(+)/K(+)ATPase activity in FW, due to up-regulation of Na(+)/K(+)ATPase activity in a significantly larger number of tubules (Student's t-test: P<0.0001). The effect of 100 nM Ang II in SW-adapted kidneys was not significant. Stimulation with increasing Ang II concentrations was performed on isolated kidney tubule cells: Ang II provoked a dose-dependent stimulation of the Na(+)/K(+)ATPase activity in FW-adapted eels, reaching a maximum at 100 nM (1.82-fold stimulation), but no significant effect was found in SW-adapted eels (ANOVA: P<0.001 and P>0.05 respectively). Isolated tubule cells stimulated with 100 nM Ang II showed a significant generation of inositol trisphosphate (InsP(3)) and an increment in calcium release from intracellular stores. In conclusion, our results suggest that tubular Na(+)/K(+)ATPase is modulated by environmental salinity, and that Ang II has a role in regulating its activity in FW-adapted eels, probably through an InsP(3)-dependent mechanism.

Angiotensin II↗

Stable isotope analyses provide new insights into ecological plasticity in a mixohaline population of European eel.

Recent studies have shown that anguillid eel populations in habitats spanning the marine-freshwater ecotone can display extreme plasticity in the range of catadromy expressed by individual fishes. Carbon and nitrogen stable isotope analysis was used to differentiate between European eels (Anguilla anguilla) collected along a short (2 km) salinity gradient ranging from <1 per thousand to approximately 30 per thousand in Lough Ahalia, a tidal Atlantic lake system. Significant differences were recorded in mean delta13C, delta15N and C:N values from eels collected from fresh, brackish and marine-dominated basins. A discriminant analysis using these predictor variables correctly classified ca. 85% of eels to salinity zone, allowing eels to be classified as freshwater (FW), brackish (BW) or marine (MW) residents. The results of the discriminant analysis also suggested that a significant proportion of eels moved between habitats (especially between FW and BW). Comparisons of several key population parameters showed significant variation between eels resident in different salinity zones. Mean condition and estimated age was significantly lower in MW eels, whilst observed length at age (a correlate of growth) was significantly higher in MW eels, intermediate in BW and lowest in FW eels. This study has demonstrated that the ecology of eels found along a short salinity gradient can be extremely plastic and that stable isotope analysis has considerable utility in demonstrating intra-population variation in diadromous fishes.

Anguilla↗

Induction of cytochrome P450 1A in the American Eel by model halogenated and non-halogenated aryl hydrocarbon receptor agonists.

Eels (Anguilla sp.) have phylogenetic, life history, and morphological characteristics which distinguish them from many other species that have been examined for cytochrome P450 1A (CYP1A) induction. Members of the family Anguillidae often occur in regions of the coastal environment that are heavily impacted by chemical contamination. Although eels have been suggested to be a useful species for biomonitoring, the sensitivity with which eel CYP1A is induced by aryl hydrocarbon receptor (AHR) agonists is not known. We investigated the dose-dependent induction of hepatic CYP1A in the American eel (Anguilla rostrata). Eels from an uncontaminated site were injected intra-peritoneally with the model AHR agonists ß-naphthoflavone (BNF), benzo[a]pyrene (B[a]P) or 3,3',4,4-tetrachlorobiphenyl (TCB) at increasing doses (BNF at 0.1, 1, 10 and 100 mg/kg, B[a]P at 0.1, 1 and 10 mg/kg, and TCB at 0.1, 1, 10 and 20 mg/kg). All three compounds produced dose-dependent induction of CYP1A content and catalytic activity. An estimated ED(50) for induction of liver microsomal EROD activity by TCB was approximately 5 mg/kg, indicating only moderate sensitivity. At comparable doses of 1 and 10 mg/kg (or 3-4 and 30-40 µmol/kg), BNF and B[a]P had 2-3-fold greater effect than TCB in eliciting hepatic CYP1A induction. Injection of radiolabeled B[a]P and TCB resulted in similar dose-dependent concentrations of these compounds in eel liver, and the hepatic inducer concentrations and CYP1A levels were correlated positively. Eels collected from New Bedford Harbor (NBH), a Superfund site highly contaminated by polycyclic aromatic hydrocarbons and polychlorinated biphenyls, had levels of microsomal CYP1A protein and EROD activity that were equivalent to the highest levels induced experimentally. Eels from less contaminated sites had correspondingly less CYP1A expression. The responses to B[a]P or BNF as compared to TCB suggest a lower efficacy and/or potency for CYP1A induction by TCB which could involve differences in the mechanisms of responses to these compounds in eels. However, the moderate sensitivity and the CYP1A induction in NBH eels support suggestions that eels may be useful in monitoring more contaminated regions.

Journal Article↗

Epidemiology of Anguillicola crassus in the European eel (Anguilla anguilla) from two rivers in southern England.

European eels Anguilla anguilla from the rivers Thames and Test, in the south of England, were examined between 2000 and 2003 for infection with the swim-bladder nematode Anguillicola crassus. Since its introduction to Thames eels at tidal estuarine locations circa 1987, A. crassus has become established in non-tidal freshwater stretches upriver and data from these locations are reported for the first time. The prevalence of infection at Thames estuary locations was higher during 2000-2003 than for the period 1987-1992. By 2003, similar prevalences were observed at freshwater and estuarine locations, but infection intensities were significantly higher in freshwater. Eels from the river Test appear to have been recently colonized by A. crassus (circa 2000). Parasite population establishment within these eels was uncharacteristically slow during 2000-2001, with low prevalence and intensity of infection, and few gravid females during this period. By 2003, infection levels in Test eels were similar to those in Thames eels. The expansion of the A. crassus population in Test eels has occurred in the absence of the paratenic host ruffe Gymnocephalus cernuus, and at suboptimal pH for the survival and infectivity of free-living larvae. The epidemiology of A. crassus in Test eels demonstrates that transmission of A. crassus by ruffe is not required for high prevalence and intensity of infection in eels. However, the consistently low and atypical levels of infection in Test eels during 2000 and 2001 suggests that paratenic transmission by ruffe may provide a substantial contribution to the dynamics of A. crassus in eels in the early years following introduction, by facilitating the rapid increase in prevalence and intensity of A. crassus infection that typically occurs during this time.

Anguilla↗

Effects of infection with Anguillicola crassus and simultaneous exposure with Cd and 3,3',4,4',5-pentachlorobiphenyl (PCB 126) on the levels of cortisol and glucose in European eel ( Anguilla anguilla ).

To investigate whether the stress response of European eels infected with Anguillicola crassus is influenced by environmental pollutants, experimentally infected eels were exposed to Cd and/or to 3,3', 4,4', 5-pentachlorobiphenyl (PCB 126). Serum cortisol and glucose concentrations of these eels were monitored over a period of 103 days and were compared with data from infected, unexposed eels as well as with data from uninfected eels. Additionally, the levels of cortisol were correlated with concentrations of Anguillicola-specific antibodies. All eels showed an initial increase of the cortisol levels until day 63. This general elevation of plasma cortisol is most likely due to handling stress, as all eels were repeatedly netted and afterwards inoculated with a feeding tube. At the end of the exposure period eels which were infected and those which were infected and simultaneously exposed to Cd and PCB showed significantly higher levels than the controls. The general course of serum glucose levels in eels resembled that of cortisol. Accordingly, Spearman correlation analysis revealed that an increase in serum cortisol concentrations is correlated with rising levels of glucose. With respect to immune-endocrine interactions a significant negative correlation between cortisol and anti-A. crassus antibodies was found. Our data show that A. crassus is the most potent stressor for European eels among the treatments tested within this study. This is important in terms of ecotoxicological studies as the main effects are caused by parasites rather than chemicals. Accordingly, effects of parasites on the physiological homeostasis of organisms must be considered in ecotoxicology. From the parasitological point of view our results suggest that probably as part of an unbalanced host-parasite interaction A. crassus evokes a strong cortisol response in A. anguilla, thereby suppressing the immune response which in turn enables the parasite to establish. The parasite-induced stress response in the newly adopted European eel might be one of the factors which contributes to the extremely effective colonizing strategy of A. crassus.

Anguilla↗

The relationship between beta-adrenoceptors and adrenergic responsiveness in trout (Oncorhynchus mykiss) and eel (Anguilla rostrata) erythrocytes.

Experiments were performed in vitro specifically to elucidate the underlying mechanism(s) of the attenuated adrenergic responses of eel (Anguilla rostrata) erythrocytes. This was achieved by comparing beta-adrenoceptor numbers and affinities in addition to (i) Na+/H+ exchange activity, (ii) cell swelling and (iii) cyclic AMP formation mediated by catecholamines in eel and trout (Oncorhynchus mykiss) erythrocytes under normoxic and hypoxic conditions. Under normoxic conditions, eel erythrocytes displayed a total absence of Na+/H+ exchange activity (as determined from measurements of extracellular pH) after addition of noradrenaline (50-1000 nmol l-1) in contrast to a pronounced dose-dependent response in trout. Incubation of the blood under hypoxic conditions, to achieve approximately 50% haemoglobin O2-saturation, further increased the extent of Na+/H+ exchange activation in trout and elicited a statistically significant, although physiologically small (10% of the response in trout), activation of H+ extrusion activity in eel. Catecholamine-mediated cell swelling, although obvious in trout, was absent in eel when estimated under hypoxic conditions. Eel erythrocytes possessed approximately 50% fewer surface beta-adrenoceptors than did trout erythrocytes, although the dissociation constants (KD) of these receptors did not differ between eel and trout. The numbers and affinities of the erythrocyte beta-adrenoceptors were not significantly affected by the hypoxic incubation. Both eel and trout erythrocytes displayed a dose-dependent elevation of cyclic AMP concentration in response to noradrenaline that was further increased by hypoxia. Surprisingly, eel erythrocytes produced larger quantities of cyclic AMP despite the lower numbers of surface beta-adrenoceptors. Thus, the absence of adrenergic swelling and the attenuated H+ extrusion response in eel erythrocytes cannot be attributed to insufficient numbers of beta-adrenoceptors or to functional uncoupling of these receptors from adenylate cyclase. Instead, the differences between trout and eel may reflect differing numbers of Na+/H+ exchangers or fundamental differences in the manner by which these exchangers are activated by cyclic AMP.

Anguilla↗

The use of the DR CALUX bioassay and indicator polychlorinated biphenyls for screening of elevated levels of dioxins and dioxin-like polychlorinated biphenyls in eel.

The DR CALUX bioassay is a very suitable screening method for dioxins and dioxin-like-PCBs in feed and food. This was, e. g. demonstrated in a survey in the Netherlands to control the dioxin levels in eel. The DR CALUX assay, but also indicator polychlorinated biphenyls (PCB) were evaluated as a screening method. Based on the limit for polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/F) [at that time 8 pg toxic equivalents (TEQ)/g eel], and the relation between PCDD/F and dioxin-like-PCB, a decision limit of 30 pg TEQ/g eel was used for screening of 153 field samples. Suspected samples (21) and part of the higher contaminated negative samples (35) were analyzed by GC/MS for dioxins, non-ortho, mono-ortho and indicator PCB, revealing 13 samples exceeding the action limit of 30 pg TEQ/g eel. Only one sample slightly exceeded the dioxin level of 8 pg TEQ/g eel. The relatively low sensitivity for mono-ortho PCB was overcome by the use of reference samples, as shown by the correlation of 0.93 between GC/MS and CALUX determined total TEQ levels. The present data show that the DR CALUX assay can be used for screening of total TEQ levels in eel. The use for dioxins only requires a safe, and therefore relatively low, decision limit. The indicator PCB also showed a good correlation with total TEQ levels, mainly due to the large contribution of the mono-ortho PCB at higher concentrations. The relation with dioxins was very poor and as such indicator PCB seem less suitable than the DR CALUX assay for screening for dioxins only. The present study clearly shows that part of the wild eel samples contains high total TEQ levels and will exceed the future European Union limit of 12 pg TEQ/g eel for dioxins and dioxin-like PCB. Especially at high TEQ levels, dioxin-like PCB contribute most to the total TEQ. In practice, wild eel presents only a minor part of the eel consumed.

Adipose Tissue↗

Atrial and ventricular natriuretic peptide concentrations in plasma of freshwater- and seawater-adapted eels.

Plasma levels of atrial and ventricular natriuretic peptide (ANP and VNP) were measured in freshwater (FW)- and seawater (SW)-adapted eels using homologous radioimmunoassays. Anesthesia with tricaine (MS222) and exposure of the pericardium for blood sampling from the ventral aorta affected plasma ANP and VNP levels. ANP and VNP levels in the ventral aorta were higher than those in the dorsal aorta when blood was collected simultaneously from both aortae of cannulated conscious FW eels, perhaps reflecting branchial extraction of the peptides. This phenomenon was not observed in SW eels. Based on these results, blood was sampled from conscious eels through a catheter implanted in the ventral aorta because it appears to reflect most accurately ANP secretion. Plasma ANP and VNP levels measured in more than 100 conscious FW and SW eels displayed large variations at each month despite acclimation to laboratory conditions at 180 for more than 1 week in FW eels or for more than 2 weeks in SW eels. However, concentrations of ANP and VNP averaged from all data were not different between FW and SW eels: 68.1 +/- 2.9 (FW: n = 155) vs 60.7 +/- 3.6 (SW: n = 104) for ANP, 51.6 +/- 3.0 (FW: n = 69) vs 55.0 +/- 3.7 (SW: n = 54) for VNP (fmol/ml in each case). Plasma Na concentration and osmolality were significantly higher in SW eels than in FW eels. Plasma ANP and VNP levels thus do not seem to be affected by chronic changes in plasma Na concentration and osmolality.

Adaptation, Physiological↗

Identification of two functional guanylin receptors in eel: multiple hormone-receptor system for osmoregulation in fish intestine and kidney.

Guanylyl cyclase C (GC-C) is a single transmembrane receptor for a family of intestinal hormones, guanylins. In the eel, we previously identified three guanylins, whose gene expression was enhanced in the intestine after transfer from fresh water to seawater. However, only limited information is available about the structure and function of their receptor(s). In the present study, we cloned full-length cDNAs encoding two isoforms of GC-C, named GC-C1 and GC-C2, from eel intestine. The predicted GC-C proteins consisted of extracellular ligand-binding domain, membrane-spanning domain, kinase-like domain and cyclase catalytic domain, in which GC-C-specific sequences were largely conserved. Phylogenetic analyses showed that the cloned membrane GCs are grouped with the GC-C of other vertebrates but not with GC-A and GC-B. However, eel GC-Cs appear to have undergone unique structural evolution compared with other GC-Cs. The three eel guanylins (guanylin, uroguanylin and renoguanylin), but not eel atrial natriuretic peptide, stimulated cGMP production dose-dependently in COS cells expressing either of the cloned cDNAs, providing functional support for assignment as eel guanylin receptors. The potency order for cGMP production was uroguanylin > guanylin > or = renoguanylin for GC-C1; guanylin > or = renoguanylin > uroguanylin for GC-C2. The distinctive ligand selectivity was consistent with the low homology (53%) of the extracellular domain of the two GC-Cs compared with that observed for other domains (74-90%). Both GC-C genes were expressed in the alimentary tract (esophagus, stomach and intestine) and kidney, and their expression was higher in the intestine of seawater-adapted eels than that of freshwater eels just as observed with the guanylin genes. However, the expression of the receptor genes was unchanged for 24h after transfer of eels from fresh water to seawater or vice versa, showing slower response of the receptors to salinity changes than their ligands. Collectively, the multiple guanylin-GC-C system may be involved as a paracrine factor in seawater adaptation at the intestine and kidney of the eel.

Amino Acid Sequence↗

Slow muscle power output of yellow- and silver-phase European eels (Anguilla anguilla L.): changes in muscle performance prior to migration.

Eels swim in the anguilliform mode in which the majority of the body axis undulates to generate thrust. For this reason, muscle function has been hypothesised to be relatively uniform along the body axis relative to some other teleosts in which the caudal fin is the main site of thrust production. The European eel (Anguilla anguilla L.) has a complex life cycle involving a lengthy spawning migration. Prior to migration, there is a metamorphosis from a yellow (non-migratory) to a silver (migratory) life-history phase. The work loop technique was used to determine slow muscle power outputs in yellow- and silver-phase eels. Differences in muscle properties and power outputs were apparent between yellow- and silver-phase eels. The mass-specific power output of silver-phase slow muscle was greater than that of yellow-phase slow muscle. Maximum slow muscle power outputs under approximated in vivo conditions were 0.24 W kg(-1) in yellow-phase eel and 0.74 W kg(-1) in silver-phase eel. Power output peaked at cycle frequencies of 0.3--0.5 Hz in yellow-phase slow muscle and at 0.5--0.8 Hz in silver-phase slow muscle. The time from stimulus offset to 90 % relaxation was significantly greater in yellow- than in silver-phase eels. The time from stimulus onset to peak force was not significantly different between life-history stages or axial locations. Yellow-phase eels shifted to intermittent bursts of higher-frequency tailbeats at a lower swimming speed than silver-phase eels. This may indicate recruitment of fast muscle at low speeds in yellow-phase eels to compensate for a relatively lower slow muscle power output and operating frequency.

Animals↗

Differential expression of neural and gonadal aromatase enzymatic activities in relation to gonadal development in Japanese eel, Anguilla japonica.

The objectives of the present study were to investigate the enzymatic characteristics of brain aromatase in Japanese eel, Anguilla japonica, as well as the correlations between aromatase activities in various tissues (brain regions, pituitary, and gonads) and ovarian development. Eel brain aromatase exhibited a K(m) of 75 nM and a V(max) of 1.14 fmol/min mg protein (91.5 fmol/h mg protein), indicating an enzymatic activity much lower than in other teleosts but similar to that in mammals. This supports the hypothesis of the occurrence of a single aromatase gene in eels (representative of an ancient group of teleosts, Elopomorphs), as in mammals, but unlike what is observed in more recent teleosts. Aromatase inhibitors could significantly suppress brain and pituitary aromatase activity. There was no significant sex difference in aromatase activity in the forebrain, midbrain, hindbrain, or pituitary, but there was in the gonads, where aromatase could be detected in the ovaries but not in the testes, in accordance with the role of this enzyme in ovarian differentiation. Comparison with another teleost, black porgy (Percomorph), under the same experimental conditions, further confirmed the low activity of aromatase in the eel. We investigated variations in brain, pituitary, and gonad aromatase activity in relation to ovarian development in control female eels (gonadosomatic index, GSI, 0.1-1.6%) as well as in eels treated with pituitary extract (experimental ovarian maturation; GSI up to 25%). Differential expression of neural and gonadal aromatase was observed in relation to the course of gonadal development. Pituitary aromatase activity increased with GSI at all stages. Brain (specially forebrain) aromatase activity significantly increased in early vitellogenic control eels (GSI>0.8%) and in treated eels. The low activity of eel aromatase may be related to the characteristic life cycle of the eel, in which there is a long delay of the onset of puberty before oceanic reproductive migration.

Androstenedione↗

Development of a radioimmunoassay for European eel growth hormone and application to the study of silvering and experimental fasting.

A specific and sensitive homologous radioimmunoassay for eel (Anguilla anguilla L.) growth hormone (angGH) has been developed. The antiserum, raised against purified angGH and used at 1:20,000 final dilution, did not cross-react with eel prolactin or thyrotropin, carp gonadotropin II, bovine GH, or serum from hypophysectornized eel. The inhibition curves for eel pituitary extracts and serum were parallel to that of angGH standard. The ED50 value was between 1 and 2 ng/tube and the recovery of purified angGH added to the serum was about 100%. In immunocytochemical studies, the antiserum, used at 1:1000 dilution, specifically labeled the somatotrophs in the pituitaries of the glass, yellow, and silver eels. The GH contents were determined in the pituitaries of glass, yellow, and silver eels and in the serum at the yellow and silver stages. GH variations during the transformation of the yellow to silver eel were examined. The results indicated a decrease in GH production between the yellow and the silver eels, possibly related to the cessation of growth at the silver stage. In contrast to the situation in the naturally fasting silver eel, submitting yellow eels to 3 months of starvation (experimental fasting) greatly increased GH production. This suggests a variation in the regulation of GH according to the type of fasting (natural or experimental) and/or the stage of the fish (yellow or silver).

Anguilla↗

Development of a homologous radioimmunoassay for eel prolactin.

A highly specific and homologous radioimmunoassay (RIA) for the measurement of prolactin (PRL) in the plasma and the pituitary of the eel was developed using a rabbit antiserum to eel PRL. PRL was purified from the pituitary of Japanese eel (Anguilla japonica). Pituitary extracts and plasma from the Japanese and European eels exhibited displacement curves parallel to the eel PRL standard. Plasma and pituitary extracts from chum salmon, rainbow trout, Japanese charr, tilapia, goldfish, and carp, as well as plasma from hypophysectomized eel, showed negligible cross-reactivity. PRL and growth hormone (GH) preparations from chum salmon, tilapia, and sheep, carp PRL, and eel GH did not cross-react with the antibody. The RIA sensitivity was less than 0.1 ng eel PRL per milliliter. Intra- and interassay coefficients of variations were 2.4 and 11.8%, respectively. The immunoreactive PRL levels in plasma and pituitary of the eel adapted to 50% seawater were significantly lower than those of the eel in fresh water. Plasma PRL levels increased maximally 2 days after transfer from seawater to fresh water, as would be expected from the well-established role of PRL in freshwater adaptation in several euryhaline teleosts.

Adaptation, Physiological↗