Bottlenose dolphin calves have multi-year elevations of plasma oxytocin compared to all other age classes.

Providing for infants nutritionally via lactation is one of the hallmarks of mammalian reproduction, and infants without motivated mothers providing for them are unlikely to survive. Mothers must maintain regular contact with infants both spatially and temporally while utilising their environment to forage, avoid threats and find shelter. However, mothers can only do this and maximise their reproductive success with some degree of co-operation from infants, despite their developing physical and cognitive capabilities. The neuropeptide hormone oxytocin (OT) triggers proximity-seeking behaviour and acts in a positive feedback loop across mother-infant bonds, stimulating appropriate pro-social behaviour across the pair. However, data on infant OT levels is lacking, and it is unclear how important infants are in maintaining mother-infant associations. The bottlenose dolphin (Tursiops truncatus) is a mammalian species that is fully physically mobile at birth and has multi-year, but individually variable, lactation periods. We investigated OT concentrations in mother-infant pairs of wild individuals compared to other age and reproductive classes. An ELISA to detect OT in dolphin plasma was successfully validated with extracted plasma. We highlight a statistical method for testing for parallelism that could be applied to other ELISA validation studies. OT concentrations were consistently elevated in calves up to at least 4 years of age with lactating mothers (12.1 ±â€¯0.9 pg/ml), while all mothers (4.5 ±â€¯0.4 pg/ml) had OT concentrations comparable to non-lactating individuals (5.9 ±â€¯0.5 pg/ml). Concentrations within infants were individually variable, and may reflect the strength of the bond with their mother. The OT system likely provides a physiological mechanism for motivating infants to perform behaviours that prevent long-term separation from their mothers during this crucial time in their life history. Elevated infant OT has also been linked to energetic and developmental advantages which may lead to greater survival rates. Environmental or anthropogenic disturbances to OT release can occur during bond formation or can disrupt the communication methods used to reinforce these bonds via OT elevation. Variation in OT expression in infants, and its behavioural and physiological consequences, may explain differences in reproductive success despite appropriate maternal behaviour expression.

High oxytocin infants gain more mass with no additional maternal energetic costs in wild grey seals (Halichoerus grypus).

Maximising infant survival requires secure attachments and appropriate behaviours between parents and offspring. Oxytocin is vital for parent-offspring bonding and behaviour. It also modulates energetic balance and neural pathways regulating feeding. However, to date the connections between these two areas of the hormone's functionality are poorly defined. We demonstrate that grey seal (Halichoerus grypus) mothers with high oxytocin levels produce pups with high oxytocin levels throughout lactation, and show for the first time a link between endogenous infant oxytocin levels and rates of mass gain prior to weaning. High oxytocin infants gained mass at a greater rate without additional energetic cost to their mothers. Increased mass gain in infants was not due to increased nursing, and there was no link between maternal mass loss rates and plasma oxytocin concentrations. Increased mass gain rates within high oxytocin infants may be due to changes in individual behaviour and energy expenditure or oxytocin impacting on tissue formation. Infancy is a crucial time for growth and development, and our findings connect the oxytocin driven mechanisms for parent-infant bonding with the energetics underlying parental care. Our study demonstrates that oxytocin release may connect optimal parental or social environments with direct physiological advantages for individual development.

Positive social behaviours are induced and retained after oxytocin manipulations mimicking endogenous concentrations in a wild mammal.

The neuropeptide hormone oxytocin modulates numerous social and parental behaviours across a wide range of species, including humans. We conducted manipulation experiments on wild grey seals (Halichoerus grypus) to determine whether oxytocin increases proximity-seeking behaviour, which has previously been correlated with endogenous oxytocin concentrations in wild seal populations. Pairs of seals that had never met previously were given intravenous injections of 0.41 µg kg-1 oxytocin or saline and were observed for 1 h post-manipulation. The dose was designed to mimic endogenous oxytocin concentrations during the observation period, and is one of the lowest doses used to manipulate behaviour to date. Seals given oxytocin spent significantly more time in close proximity to each other, confirming that oxytocin causes conspecifics to seek others out and remain close to one another. Aggressive and investigative behaviours also significantly fell after oxytocin manipulations. Despite using a minimal oxytocin dose, pro-social behavioural changes unexpectedly persisted for 2 days despite rapid dose clearance from circulation post-injection. This study verifies that oxytocin promotes individuals staying together, demonstrating how the hormone can form positive feedback loops of oxytocin release following conspecific stimuli, increased motivation to remain in close proximity and additional oxytocin release from stimuli received while in close proximity.

Myo-inositol phosphate synthase expression in the European eel (Anguilla anguilla) and Nile tilapia (Oreochromis niloticus): effect of seawater acclimation.

A single MIPS gene (Isyna1/Ino1) exists in eel and tilapia genomes with a single myo-d-inositol 3-phosphate synthase (MIPS) transcript identified in all eel tissues, although two MIPS spliced variants [termed MIPS(s) and MIPS(l)] are found in all tilapia tissues. The larger tilapia transcript [MIPS(l)] results from the inclusion of the 87-nucleotide intron between exons 5 and 6 in the genomic sequence. In most tilapia tissues, the MIPS(s) transcript exhibits much higher abundance (generally >10-fold) with the exception of white skeletal muscle and oocytes, in which the MIPS(l) transcript predominates. SW acclimation resulted in large (6- to 32-fold) increases in mRNA expression for both MIPS(s) and MIPS(l) in all tilapia tissues tested, whereas in the eel, changes in expression were limited to a more modest 2.5-fold increase and only in the kidney. Western blots identified a number of species- and tissue-specific immunoreactive MIPS proteins ranging from 40 to 67 kDa molecular weight. SW acclimation failed to affect the abundance of any immunoreactive protein in any tissue tested from the eel. However, a major 67-kDa immunoreactive protein (presumed to be MIPS) found in tilapia tissues exhibited 11- and 54-fold increases in expression in gill and fin samples from SW-acclimated fish. Immunohistochemical investigations revealed specific immunoreactivity in the gill, fin, skin, and intestine taken from only SW-acclimated tilapia. Immunofluorescence indicated that MIPS was expressed within gill chondrocytes and epithelial cells of the primary filaments, basal epithelial cell layers of the skin and fin, the cytosol of columnar intestinal epithelial and mucous cells, as well as unknown entero-endocrine-like cells.

Maternal Oxytocin Is Linked to Close Mother-Infant Proximity in Grey Seals (Halichoerus grypus).

Maternal behaviour is a crucial component of reproduction in all mammals; however the quality of care that mothers give to infants can vary greatly. It is vital to document variation in maternal behaviour caused by the physiological processes controlling its expression. This underlying physiology should be conserved throughout reproductive events and should be replicated across all individuals of a species; therefore, any correlates to maternal care quality may be present across many individuals or contexts. Oxytocin modulates the initiation and expression of maternal behaviour in mammals; therefore we tested whether maternal plasma oxytocin concentrations correlated to key maternal behaviours in wild grey seals (Halichoerus grypus). Plasma oxytocin concentrations in non-breeding individuals (4.3 ± 0.5 pg/ml) were significantly lower than those in mothers with dependent pups in both early (8.2 ± 0.8 pg/ml) and late (6.9 ± 0.7 pg/ml) lactation. Maternal plasma oxytocin concentrations were not correlated to the amount of nursing prior to sampling, or a mother's nursing intensity throughout the dependent period. Mothers with high plasma oxytocin concentrations stayed closer to their pups, reducing the likelihood of mother-pup separation during lactation which is credited with causing starvation, the largest cause of pup mortality in grey seals. This is the first study to link endogenous oxytocin concentrations in wild mammalian mothers with any type of maternal behaviour. Oxytocin's structure and function is widely conserved across mammalian mothers, including humans. Defining the impact the oxytocin system has on maternal behaviour highlights relationships that may occur across many individuals or species, and such behaviours heavily influence infant development and an individual's lifetime reproductive success.

Conspecific recognition and aggression reduction to familiars in newly weaned, socially plastic mammals.

Recognising conspecifics and behaving appropriately towards them is a crucial ability for many species. Grey seals (Halichoerus grypus) show varying capabilities in this regard: mother-pup recognition has been demonstrated in some geographical populations but is absent in others, yet there is evidence that individuals aggregate with prior associates. The recognition capabilities of newly weaned grey seal pups were investigated using class recognition trials within the habituation/dishabituation paradigm. Trials took place in pens, using pairs of individuals that either had previously cohabited (familiar) or that had never met before (stranger). Frequencies of olfactory and visual investigative behaviours ('checks') and aggressive interactions were recorded during trials. Familiar individuals recognised each other: paired strangers showed significantly more checks and aggressive interactions than were seen in trials pairing familiars. Oxytocin concentrations in post-trial plasma samples were analysed to investigate the underlying physiology modulating recognition abilities; however, no significant differences were detected between familiar or stranger trials. This study demonstrates that at a young age, grey seals can recognise individuals they have previously encountered. Recognition abilities in this species have adaptive value by allowing the reduction of costly aggressive interactions between familiar conspecifics, which is often cited as the first step towards the evolution of sociality in a species. This study is the first with wild subjects to find conspecific recognition abilities in a pinniped species outside of reproductive contexts. It demonstrates that even largely solitary species can be capable of recognition and pro-social behaviours that benefit them during times when they must aggregate.

Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure.

BACKGROUND: The neuropeptide oxytocin is increasingly the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHOD: This study determined the sample collection and analysis protocols that cause the least amounts of protocol dependant variation in plasma oxytocin concentrations detected by ELISA. The effect of vacutainer type, sample extraction prior to analysis and capture and restraint protocol were investigated while validating an assay protocol for two novel species, grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina). RESULTS: Where samples are extracted prior to analysis, vacutainer type (EDTA mean: 8.25±0.56 pg/ml, heparin mean: 8.25±0.62 pg/ml, p=0.82), time taken to obtain a sample and restraint protocol did not affect the concentration of oxytocin detected. However, concentrations of oxytocin detected in raw plasma samples were significantly higher than those in extracted samples, and varied significantly with vacutainer type (EDTA mean: 534.4±43.7 pg/ml, heparin mean: 300.9±19.6 pg/ml, p<0.001) and capture and restraint methodology. There was no relationship between oxytocin concentrations detected in raw and extracted plasma (p=0.25). COMPARISON WITH EXISTING METHOD(S): Over half the reviewed published studies analysing plasma oxytocin use raw plasma and different vacutainer types are used without consistency or justification through-out the literature. CONCLUSIONS: We caution that studies using raw plasma are likely to over estimate oxytocin concentrations, cannot be used to accurately infer true values via correlations and are susceptible to variation according vacutainer type.

Seawater acclimation and inositol monophosphatase isoform expression in the European eel (Anguilla anguilla) and Nile tilapia (Orechromis niloticus).

Inositol monophosphatase (IMPA) is responsible for the synthesis of inositol, a polyol that can function as an intracellular osmolyte helping re-establish cell volume when exposed to hypertonic environments. Some epithelial tissues in euryhaline teleosts such as the eel and tilapia encounter considerable hyperosmotic challenge when fish move from freshwater (FW) to seawater (SW) environments; however, the roles played by organic osmolytes, such as inositol, have yet to be determined. Syntenic analysis has indicated that, as a result of whole genome- and tandem-duplication events, up to six IMPA isoforms can exist within teleost genomes. Four isoforms are homologs of the mammalian IMPA1 gene, and two isoforms are homologs of the mammalian IMPA2 gene. Although the tissue-dependent isoform expression profiles of the teleost isoforms appear to be species-specific, it was primarily mRNA for the IMPA1.1 isoform that was upregulated in epithelial tissues after fish were transferred to SW (up to 16-fold in eel and 90-fold in tilapia). Although up-regulation of IMPA1.1 expression was evident in many tissues in the eel, more substantial increases in IMPA1.1 expression were found in tilapia tissues, where SW acclimation resulted in up to 2,000-fold increases in protein expression, 16-fold increases in enzyme activity and 15-fold increases in tissue inositol contents. Immunohistochemical studies indicated that the tissue and cellular distribution of IMPA1.1 protein differed slightly between eels and tilapia; however, in both species the basal epithelial cell layers within the skin and fin, and the branchial epithelium and interstitial cells within the kidney, exhibited high levels of IMPA1.1 protein expression.

Transcriptomic approach to the study of osmoregulation in the European eel Anguilla anguilla.

In euryhaline teleosts, osmoregulation is a fundamental and dynamic process that is essential for the maintenance of ion and water balance, especially when fish migrate between fresh water (FW) and sea water (SW) environments. The European eel has proved to be an excellent model species to study the molecular and physiological adaptations associated with this osmoregulatory plasticity. The life cycle of the European eel includes two migratory periods, the second being the migration of FW eels back to the Sargasso Sea for reproduction. Various anatomical and physiological changes allow the successful transition to SW. The aim of this study was to use a microarray approach to screen the osmoregulatory tissues of the eel for changes in gene expression following acclimation to SW. Tissues were sampled from fish at selected intervals over a 5-mo period following FW/SW transfer, and RNA was isolated. Suppressive subtractive hybridization was used for enrichment of differentially expressed genes. Microarrays comprising 6,144 cDNAs from brain, gill, intestine, and kidney libraries were hybridized with appropriate targets and analyzed; 229 differentially expressed clones with unique sequences were identified. These clones represented the sequences for 95 known genes, with the remaining sequences (59%) being unknown. The results of the microarray analysis were validated by quantification of 28 differentially expressed genes by Northern blotting. A number of the differentially expressed genes were already known to be involved in osmoregulation, but the functional roles of many others, not normally associated with ion or water transport, remain to be characterized.

Body fluid volume regulation in elasmobranch fish.

This review addresses an often overlooked aspect of elasmobranch osmoregulation, i.e., control of body fluid volume. More specifically the review addresses the impact of changes in blood volume in elasmobranchs exposed to different environmental salinities. Measurement of blood volume in the European lesser-spotted dogfish, Scyliorhinus canicula, following acute and chronic exposure to 80% and 120% seawater (SW) is reported. In 80%, 100% and 120% SW-adapted S. canicula, blood volume was 6.3+/-0.2, 5.6+/-0.2 and 4.6+/-0.2 mL 100 g(-1) body mass, respectively. Blood volume was significantly higher and lower in 80% and 120% SW-acclimated animals compared to 100% SW controls. Comparisons are made between these results and previously published data. The role of drinking and volume regulation in elasmobranchs is discussed. For the first time measured water reabsorption rates and solute flux rates across the elasmobranch intestinal epithelia are presented. Water reabsorption rates did not differ between 100% SW-adapted bamboo shark, Chiloscyllium plagiosum, and fish acutely transferred to 140% SW. For the most part net solute flux rates and direction for both the 100% and 140% SW groups were the same with the exception of a net efflux of chloride and potassium in the 140% group and influx of these ions in the 100% adapted group. The significance of the intestine as part of the overall elasmobranch osmoregulatory strategy is discussed as is the role of the kidneys, rectal gland and gills in the regulation of body fluid volume in this class of vertebrates.

The effects of freshwater to seawater transfer on circulating levels of angiotensin II, C-type natriuretic peptide and arginine vasotocin in the euryhaline elasmobranch, Carcharhinus leucas.

This study examined the effect of transfer to increased environmental salinity on the circulating levels of angiotensin II (ANG II), C-type natriuretic peptide (CNP), and arginine vasotocin (AVT) in the euryhaline elasmobranch, Carcharhinus leucas. Plasma levels of ANG II and CNP were significantly increased in C. leucas chronically acclimated to seawater (SW) in comparison to freshwater (FW) acclimated fish. There was no difference in plasma AVT levels. Acute transfer of FW fish to 75% SW induced an increase in plasma ANG II levels within 12 h, and subsequent transfer from 75 to 100% SW further increased plasma ANG II levels at both 24 and 72 h. No change in plasma CNP was observed during acute transfer to increased salinity. However, a significant increase in plasma AVT levels was observed following 96 h in 75% SW and 24 h in 100% SW. In chronically SW acclimated C. leucas plasma osmolality, sodium, chloride, and urea were all significantly higher than FW acclimated fish but there was no difference in haematocrit. Acute transfer of C. leucas to 75% SW induced a significant increase in plasma osmolality, sodium and urea concentrations within 96 h of transfer. Subsequent transfer from 75 to 100% SW induced a further increase in these variables within 24 h in addition to a significant increase in plasma chloride above control levels. Haematocrit did not differ between the experimental and control groups throughout the acute study. Circulating levels of ANG II were significantly correlated to plasma, sodium, chloride, and urea concentrations during acclimation to SW. Conversely, circulating levels of CNP and AVT did not correlate to plasma osmolytes, however, CNP was significantly correlated to haematocrit during acclimation to seawater.

Effects of angiotensin II and C-type natriuretic peptide on the in situ perfused trunk preparation of the dogfish, Scyliorhinus canicula.

The renal roles of physiologically relevant doses of angiotensin II (Ang II) and C-type natriuretic peptide (CNP) were investigated in the dogfish, Scyliorhinus canicula, using an in situ perfused trunk preparation. Perfusion with 10(-9) M Ang II resulted in a glomerular antidiuresis and decreases in perfusate flow rate, transport maxima for glucose and the proportion of filtering glomeruli. In addition, the renal clearances and excretion of urea, sodium, and chloride were significantly reduced, whereas the relative clearances of these parameters remained unchanged. In contrast, perfusion of 10(-9) M CNP caused a glomerular diuresis, an increase in transport maxima for glucose, but no significant change in the proportion of filtering glomeruli. In addition, the renal clearances of urea, sodium, and chloride were significantly increased but there was no effect on the relative clearances of urea, sodium, or chloride. Perfusion with 10(-10) M Ang II or CNP had no significant renal effects. Our results suggest that these hormones act at the level of the glomeruli rather than at a tubular level.

Hepatic urea biosynthesis in the euryhaline elasmobranch Carcharhinus leucas.

Plasma urea levels and hepatic urea production in the euryhaline bull shark, Carcharhinus leucas, acclimated to freshwater and seawater environments were measured. It was found that plasma urea concentration increased with salinity and that this increase was, in part, the result of a significant increase in hepatic production of urea. This study provides direct evidence that hepatic production of urea plays an important role in the osmoregulatory strategy of C. leucas.

Sequence, circulating levels, and expression of C-type natriuretic peptide in a euryhaline elasmobranch, Carcharhinus leucas.

The present study has examined expression and circulating levels of C-type natriuretic peptide (CNP) in the euryhaline bull shark, Carcharhinus leucas. Complementary DNA and deduced amino acid sequence for CNP in C. leucas were determined by RACE methods. Homology of CNP amino acid sequence in C. leucas was high both for proCNP and for mature CNP when compared with previously identified elasmobranch CNPs. Mature CNP sequence in C. leucas was identical to that in Triakis scyllia and Scyliorhinus canicula. Levels of expression of CNP mRNA were significantly decreased in the atrium but did not change in either the brain or ventricle following acclimation to a SW environment. However, circulating levels of CNP significantly increased from 86.0+/-7.9 fmol ml(-1) in FW to 144.9+/-19.5 fmol ml(-1) in SW. The results presented demonstrate that changes in environmental salinity influences both synthesis of CNP from the heart and also circulating levels in C. leucas. Potential stimulus for release and modes of action are discussed.

Effect of cortisol on aquaporin expression in the esophagus of the European eel, Anguilla anguilla.

Long-term cortisol infusion into freshwater (FW)-adapted eels induced a significant increase in aquaporin-1 (AQP1) mRNA expression within the esophageal epithelium of migratory "silver" eels, but not in nonmigratory, immature "yellow" eels. Cortisol treatment had no significant effect on the mRNA abundance of a second aquaporin-1 isoform, termed AQP1dup, which exhibited a highly variable expression profile among individual members of all fish groups. These results suggest that cortisol, at plasma concentrations similar to that found during FW/seawater (SW) acclimation, induces upregulation in AQP1 expression and thus increases esophageal water permeability during the migration of eels to the SW environment.

Glomerular effects of AVT on the in situ perfused trunk preparation of the dogfish.

The effects of arginine vasotocin on the pattern of glomerular perfusion in an elasmobranch fish were examined using an in situ perfused renal trunk preparation. A significant antidiuresis was coupled with a marked reduction in the proportion of filtering glomeruli (86%-25%) and an increase in the proportion of perfused but not filtering (10%-53%) and non-perfused glomeruli (4%-22%). However, the reduction in filtering glomeruli was greater than predicted by measurements of urine flow rate and glomerular filtration rate alone, suggesting that alterations in single nephron glomerular filtration rate may occur.

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.

Sex-biased investment in yolk androgens depends on female quality and laying order in zebra finches (Taeniopygia guttata).

The Trivers-Willard hypothesis predicts sex biases in parental investment according to parental condition. In addition, parents may need to sex bias their investment if there is an asymmetry between the sexes in offspring fitness under different conditions. For studying maternal differential investment, egg resources are ideal subjects because they are self contained and allocated unequivocally by the female. Recent studies show that yolk androgens can be beneficial to offspring, so here we test for sex-biased investment with maternal investment of yolk testosterone (T) in zebra finch (Taeniopygia guttata) eggs. From the Trivers-Willard hypothesis, we predicted females to invest more in male eggs in optimum circumstances (e.g. good-condition mother, early-laid egg), and more in female eggs under suboptimal conditions (e.g. poor-condition mother, late-laid egg). This latter prediction is also because in this species there is a female nestling disadvantage in poor conditions and we expected mothers to help compensate for this in female eggs. Indeed, we found more yolk T in female than male eggs. Moreover, in accordance with our predictions, yolk T in male eggs increased with maternal quality relative to female eggs, and decreased with laying order relative to female eggs. This supports our predictions for the different needs and value of male and female offspring in zebra finches. Our results support the idea that females may use yolk androgens as a tool to adaptively manipulate the inequalities between different nestlings.

Regulation of expression of two aquaporin homologs in the intestine of the European eel: effects of seawater acclimation and cortisol treatment.

Complementary DNAs encoding homologs of the mammalian aquaglyceroporins (termed AQPe) and aquaporin-1 isoforms (termed AQP1) were isolated from the European eel. The AQP amino acid sequences share 35-54% identity with other known human AQPs. Although AQPe mRNA expression was approximately equivalent along the entire length of the gut, AQP1 expression was the highest in the posterior/rectal segment. Seawater (SW) acclimation increased AQP1 mRNA abundance by 5- and 17-fold in the anterior, 14- and 23-fold in the mid-, and 9- and 7-fold in the posterior/rectal gut regions of yellow and silver eels, respectively. SW acclimation had an effect on AQPe mRNA expression only in the midintestine of silver eels, where a small but significant 1.7-fold increase in abundance was measured. Western blots using an eel AQP1-specific antibody identified the presence of a major immunoreactive 28-kDa protein, primarily within the posterior/rectal segment. A 3-wk SW transfer induced an increase in AQP1 protein abundance in all intestinal segments, with the posterior/rectal region still expressing protein levels approximately 40- and 8-fold higher than the anterior and midsegments, respectively. Strong AQP1 immunofluorescence was detected within the vascular endothelium in both freshwater (FW)- and SW-acclimated eels and in the epithelial apical brush border in the posterior/rectal gut regions of SW-acclimated eels. Cortisol infusion into FW eels had no effect on intestinal AQPe mRNA expression but induced increases in AQP1 mRNA and protein levels. These results provide evidence for the presence of a SW-induced and steroid-regulated AQP water channel pathway within the intestine of the European eel.