Changes in vision and olfaction during sexual maturation in the European eel Anguilla anguilla (L.)

Retinal structure was examined in sexually immature and artificially matured female Anguilla anguilla . Inner nuclear layer cell numbers decreased from 600 mm⁻¹ retinal cross-section to 300 mm^−I and ganglion cells from 60 to 30 mm⁻¹ cross-section, during sexual maturation. Most of the decrease occurred prior to the stage of maturation at which migration begins. Electroretinograms were recorded from the intact eyes of immature and maturing eels. There was no change in scotopic sensitivities to light of wavelengths 480 and 520 nm, with increasing sexual maturity. Olfactory organs were examined in female eels of a similar range of maturity states and were found to atrophy in artificially matured eels of advanced development. The density of mucous cells in olfactory lamellae decreased from a maximum of 443 mm⁻² in sexually immature eels to as low as 19 mm⁻² in sexually maturing eels. The changes in vision and olfaction were thought to indicate a change in the relative importance of the two sensory modalities with sexual maturation.     

Changes in body musculature with sexual maturation in the European eel, Anguilla anguilla (L.)

The structure and distribution of red and white muscles have been investigated in female Anguilla anguilla of varying sexual maturity. Red muscle volumes increased with sexual maturation from approximately 5% of total muscle volume in sexually immature eels to a maximum of 13.3% in sexually maturing animals. Volume increase was due to increasing fibre diameter rather than recruitment of new fibres. Intracellular volume fractions of lipid and mitochondria increased markedly in maturing fish. Artificially matured cels showed the largest increases in muscle lipid. The possible functional roles of red and white muscle during migration are discussed.     

Changes in the gene expression profiles of the brains of male European eels (Anguilla anguilla) during sexual maturation

Background

The vertebrate brain plays a critical role in the regulation of sexual maturation and reproduction by integrating environmental information with developmental and endocrine status. The European eel Anguilla anguilla is an important species in which to better understand the neuroendocrine factors that control reproduction because it is an endangered species, has a complex life cycle that includes two extreme long distance migrations with both freshwater and seawater stages and because it occupies a key position within the teleost phylogeny. At present, mature eels have never been caught in the wild and little is known about most aspects of reproduction in A. anguilla. The goal of this study was to identify genes that may be involved in sexual maturation in experimentally matured eels. For this, we used microarrays to compare the gene expression profiles of sexually mature to immature males.

Results

Using a false discovery rate of 0.05, a total of 1,497 differentially expressed genes were identified. Of this set, 991 were expressed at higher levels in brains (forebrain and midbrain) of mature males while 506 were expressed at lower levels relative to brains of immature males. The set of up-regulated genes includes genes involved in neuroendocrine processes, cell-cell signaling, neurogenesis and development. Interestingly, while genes involved in immune system function were down-regulated in the brains of mature males, changes in the expression levels of several receptors and channels were observed suggesting that some rewiring is occurring in the brain at sexual maturity.

Conclusions

This study shows that the brains of eels undergo major changes at the molecular level at sexual maturity that may include re-organization at the cellular level. Here, we have defined a set of genes that help to understand the molecular mechanisms controlling reproduction in eels. Some of these genes have previously described functions while many others have roles that have yet to be characterized in a reproductive context. Since most of the genes examined here have orthologs in other vertebrates, the results of this study will contribute to the body of knowledge concerning reproduction in vertebrates as well as to an improved understanding of eel biology.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-799) contains supplementary material, which is available to authorized users.     

Changes in the skin-scale complex with sexual maturation in the European eel, Anguilla anguilla (L.)

Structural features of the epidermis, dermis and scales were examined in adult female Anguilla anguilla of varying sexual maturity. Advanced sexual stages were obtained by hormone injection. The dermis increased in thickness with both size and sexual development. Epidermal thickness was unrelated to size or maturity but mucous cell densities decreased in more mature animals. Hormone treated eels showed a loss of mucous cells accompanied by degeneration of the epidermis. Scale areas increased from 50% in sexually immature adults to 145% in sexually maturing eels. Skin calcium values ranged from 0.27 to 0.83 mg cm⁻² and did not show any relation to either scale or sexual maturity. The possible relevance of structural changes to migration is discussed.     

Relation of visual changes to the onset of sexual maturation in the European eel Anguilla anguilla (L.)

Eye size was correlated with body length and gonad development in 112 untreated and 33 hormone injected female Anguilla anguilla . A length-related measure of eye size was developed as an index of sexual maturity. Eels of eye index of 6.5 or less were classed as sexually immature adults , those of < 6.5 as sexually Maturing adults . Structural changes in the eye during maturation were examined. Total numbers of rods increased markedly while density of photoreceptors remained approximately constant. Cone density decreased with degenerative changes occurring. Changes in the eye were found to be essentially complete early in the maturation process.     

A pseudocystic lipoma in the European eel (Anguilla anguilla)

This is the first report of lipoma in the European eel Anguilla anguilla. In a single eel that was obtained from a polyculture fish farm in northern Egypt, a yellowish swelling (10 mm in diameter) was observed near the tip of the lower lip. The tumor was composed of mature lipocytes that occasionally fused into an unlined cystic space.     

Development of the swimbladder in the European eel (Anguilla anguilla)

The swimbladder of the adult eel, Anguilla anguilla, with its bipolar countercurrent system, the rete mirabile, is a widely used model for swimbladder function, but very little is known about the development of this swimbladder. Our histological studies on the developing swimbladder revealed that during metamorphosis the swimbladder becomes present as a dorsal outgrowth of the esophagus. It is filled with surfactant, and gas was not detected in the swimbladder. In the young glass-eel, the epithelial (gas gland) cells of the swimbladder are columnar, but do not yet have the typical basolateral labyrinth established in adult animals. Few blood vessels are found in the swimbladder tissue, and the submucosa is present as a thick layer of connective tissue, giving a large diffusion distance between blood vessel and swimbladder lumen. Within the next 2 or 3 months of development, gas gland cells develop their typical basolateral labyrinth, and the thickness of the submucosa is significantly reduced, resulting in a short diffusion distance between blood vessels and the swimbladder lumen. The first filling of the swimbladder with gas is observed while the gas gland cells are still in a poorly differentiated status and it appears unlikely that these cells can accomplish their typical role in gas deposition. The presence of small gas bubbles in the swimbladder as well as in the ductus pneumaticus at the time of initial swimbladder inflation suggests that the swimbladder is filled by air gulping or possibly by taking up gas bubbles from the water.     

Cloning and Expression of Guanylin from the European eel (Anguilla anguilla)

Extracts of intestinal epithelia from the European eel (Anguilla anguilla) stimulated cGMP production in the T84 human colon carcinoma cell line which suggested the presence of a guanylin-like peptide in this teleost fish. Degenerate oligonucleotide primers were subsequently used in RT-PCR resulting in the amplification, cloning, and sequencing of two cDNAs which represent possible 5' spliceoforms of an eel homologue of the mammalian peptide, guanylin. Northern blotting indicated that the main site of expression of the eel peptide is in the intestine with much lower signals also detected in the kidney. Intestinal expression of guanylin mRNA is up-regulated in both nonmigratory "yellow" and the more sexually mature, migratory "silver" eels following acclimation to the seawater environment. These results suggest that this peptide signalling system may play a role in osmoregulation in euryhaline teleost fish during migration between the marine and freshwater environments.     

Replication banding in the chromosomes of the European eel (Anguilla anguilla)

The chromosomes of the European eel Anguilla anguilla have been analyzed with a replication banding technique from lymphocyte cultures treated with 5-BrdU. This technique allows us to identify with high resolution the individual chromosome pairs and to differentiate classes of chromatin by the order of replication. The replication banding obtained on the chromosomes of European eel can be related with the structural bands described in this species.     

Hearing in the eel (Anguilla anguilla)

Summary The auditory sensitivity in the European eel (Anguilla anguilla) was measured using an acoustic tube producing sound stimuli with different ratios between sound pressure and particle motion. The upper audible frequency limit in the eel was about 300 Hz. At low frequencies the relevant stimulus parameter was particle motion, excluding involvement of the swimbladder. At the higher frequencies within the audible range the swimbladder conveyed an auditory advantage for stimuli with a high ratio between pressure and particle motion. The eel has an extremely long distance between the swimbladder and the ear. An auditory function of the swimbladder in this species therefore indicates an efficient transmission channel for the reradiated swimbladder pulsations between the bladder and the ear, although specialized anatomical adaptations for this purpose are lacking.     

A checklist of the protozoan and metazoan parasites of European eel (Anguilla anguilla): checklist of Anguilla anguilla parasites

A comprehensive literature review on prevalence and distribution of parasites of the European eel (Anguilla anguilla) in European waters was conducted to set a baseline for future trend analyses and biodiversity considerations. The resulting checklist compiled from peer reviewed journals as well as grey literature including scientific reports, conference contributions, PhD theses and own survey data includes all protozoan and metazoan parasites of the European eel from freshwater, brackish and marine habitats. Parasites are arranged according to phylum and family. Species within families are listed alphabetically. Infected host organs, recorded habitats, localities, countries and literature citations are provided for each species. A total of 161 parasite species ? taxa from 30 countries are listed. With the trypanorhynch cestode Tentacularia coryphaenae a new host record could be documented. Furthermore, three new locality records of parasitic nematodes could be added based on own survey data: Anisakis simplex from an eel caught in the Baltic Sea, Eustrongylides mergorum for German freshwaters, and Goezia anguillae from the Tagus estuary in Portugal. Special consideration was given to the distribution of the swimbladder nematode Anguillicola crassus. A map and table of first reported records of this neozoan parasite species for 25 countries as well as a survey map indicating all published localities documented in the checklist are provided.     

Iron-rich particles in European eel (Anguilla anguilla L.)

Magnetic material in the European eel (Anguilla anguilla L.) was investigated by a combination of magnetic susceptibility measurements, energy dispersive X-ray fluorescence analysis and transmission electron microscopy. It was shown that the magnetic material is associated with iron. The main part of the iron is present in the form of iron-rich particles with irregular shapes about 100-3000 A large. The structures of magnetite (Fe3O4), hematite (alpha-Fe2O3) and alpha-iron (bcc structure) were identified. The particles are composed of more than one of these phases with magnetite being a minority phase when present. The iron-rich particles found in the eel are different from the materials reported for bacteria or bees.     

Response of seaward-migrating European eel (Anguilla anguilla) to manipulated flow fields

Anthropogenic structures (e.g. weirs and dams) fragment river networks and restrict the movement of migratory fish. Poor understanding of behavioural response to hydrodynamic cues at structures currently limits the development of effective barrier mitigation measures. This study aimed to assess the effect of flow constriction and associated flow patterns on eel behaviour during downstream migration. In a field experiment, we tracked the movements of 40 tagged adult European eels (Anguilla anguilla) through the forebay of a redundant hydropower intake under two manipulated hydrodynamic treatments. Interrogation of fish trajectories in relation to measured and modelled water velocities provided new insights into behaviour, fundamental for developing passage technologies for this endangered species. Eels rarely followed direct routes through the site. Initially, fish aligned with streamlines near the channel banks and approached the intake semi-passively. A switch to more energetically costly avoidance behaviours occurred on encountering constricted flow, prior to physical contact with structures. Under high water velocity gradients, fish then tended to escape rapidly back upstream, whereas exploratory ‘search’ behaviour was common when acceleration was low. This study highlights the importance of hydrodynamics in informing eel behaviour. This offers potential to develop behavioural guidance, improve fish passage solutions and enhance traditional physical screening.     

Reflex bradycardia does not influence oxygen consumption during hypoxia in the European eel (Anguilla anguilla)

Most teleost fish reduce heart rate when exposed to acute hypoxia. This hypoxic bradycardia has been characterised for many fish species, but it remains uncertain whether this reflex contributes to the maintenance of oxygen uptake in hypoxia. Here we describe the effects of inhibiting the bradycardia on oxygen consumption (MO₂), standard metabolic rate (SMR) and the critical oxygen partial pressure for regulation of SMR in hypoxia (Pcrit) in European eels Anguilla anguilla (mean ± SEM mass 528 ± 36 g; n = 14). Eels were instrumented with a Transonic flow probe around the ventral aorta to measure cardiac output (Q) and heart rate (f _H). MO₂ was then measured by intermittent closed respirometry during sequential exposure to various levels of increasing hypoxia, to determine Pcrit. Each fish was studied before and after abolition of reflex bradycardia by intraperitoneal injection of the muscarinic antagonist atropine (5 mg kg⁻¹). In the untreated eels, f _H fell from 39.0 ± 4.3 min⁻¹ in normoxia to 14.8 ± 5.2 min⁻¹ at the deepest level of hypoxia (2 kPa), and this was associated with a decline in Q, from 7.5 ± 0.8 mL min⁻¹ kg⁻¹ to 3.3 ± 0.7 mL min⁻¹ kg⁻¹ in normoxia versus deepest hypoxia, respectively. Atropine had no effect on SMR, which was 16.0 ± 1.8 μmol O₂ kg⁻¹ min⁻¹ in control versus 16.8 ± 0.8 μmol O₂ kg⁻¹ min⁻¹ following treatment with atropine. Atropine also had no significant effect on normoxic f _H or Q in the eel, but completely abolished the bradycardia and associated decline in Q during progressive hypoxia. This pharmacological inhibition of the cardiac responses to hypoxia was, however, without affect on Pcrit, which was 11.7 ± 1.3 versus 12.5 ± 1.5 kPa in control versus atropinised eels, respectively. These results indicate, therefore, that reflex bradycardia does not contribute to maintenance of MO₂ and regulation of SMR by the European eel in hypoxia.     

Colonisation of freshwater habitats by the European eel Anguilla anguilla

1. The spatial distribution of European eels in 18 U.K. rivers was related to distance from tidal limit using a negative exponential model. This function accounted for between 19 and 90% of the variation in eel density where quantitative data was available. For semiquantitative data the negative exponential function was a significant predictor of eel densities in only six out of 10 cases, although all rivers showed a consistent decline in abundance with distance upstream from the tidal limit. 2. The spatial distribution of different age groups of European eel in River Severn showed an initial rapid dispersion into freshwater followed by a much slower dispersion rate. Movement of the population upstream by a wave‐form migration process does not occur in this system. Instead colonisation of freshwaters can be seen as a two‐phase dispersion. Phase‐1 is a rapid dispersion upstream driven by density at the point source. Phase‐2 commences once the eels become yellow eels and is equivalent to random diffusion of particles. 3. These processes have important implications for the penetration of freshwaters with reduced numbers of eel larvae arriving on the coast of Europe and North America. Eel abundance will decrease more in freshwaters in an upstream direction whilst it may remain stable or decrease to a lesser extent in estuaries. They are also able to explain the demography of eels migrating upstream over weirs and the observations of varying sex ratios within catchments. We conclude that a dispersion model dependent on age, temperature, difficulty of migration, habitat quality and density of eels should be an important part of freshwater eel management.     

Swimming performance of European eel (Anguilla anguilla (L.)) elvers

To determine the relation between swimming endurance time and burst swimming speed, elvers of the European eel, Anguilla anguilla (L.), were made to swim at speeds from 3.6 to 7.2 L (body lengths) s⁻¹ in both fresh and sea water. Swimming endurance time of elvers averaging 7.2 cm total length decreased logarithmically with increased swimming speed from 3.0 min at 3.5 L s⁻¹ to 0.7 min at 5.0 L s⁻¹, and again logarithmically but with a lesser slope to 0.27 min at 7.5 L s⁻¹. No differences were found between fresh and sea water elvers. In still water, elvers could swim at high speeds for about 10–45m before exhaustion, depending upon speed. Elvers would be able to make virtually no progress against water currents >50 cm s⁻¹. Drift in coastal water currents and selective tidal transport probably involve swimming speeds below those tested in this study. Migration into freshwater streams undoubtedly involves avoidance of free stream speeds and a combination of burst and sustained swimming.     

Ovarian metabolic pathways of steroid biosynthesis in the European eel (Anguilla anguilla L.) at the silver stage

Ovarian slices of the European eel at the silver stage were incubated with 4 tritiated precursors (pregnenolone, progesterone, androstenedione, testosterone) in the presence or not of an inhibitor of 11β-hydroxylase activity, metopirone. Ether extracts were submitted to a gradient elution chromatography on celite columns. Isolated peaks were identified by isopolarity on TLC, microchemical reactions and recrystallization to constant specific activity. Interpretation of the results shows that the ovary of the European eel contains the following enzymes: a 3β-hydroxysteroid dehydrogenase, 5→4-ene-isomerase complex, a 17α-hydroxylase, a C₂₁-C₁₉ desmolase, a 17β-hydroxysteroid oxidoreductase, a 5α-reductase, a 3β-hydroxysteroid oxidoreductase and an aromatase complex. Metopirone effect indicates the presence of an 11β-hydroxylase activity. At this stage, 5β-reductase, 20β-reductase and 21-hydroxylase activities are not detected in the ovary. Water-soluble steroids were formed from all the precursors used. It appears that the ovarian biosynthesis is orientated towards the production of 5α-reduced compounds and that this might limit the production of 17β-estradiol by lowering the amount of disposable endogenous precursors.