Gill Permeability Research Articles

Effects of short-term exposure to the heavy metal, nickel chloride (Nicl2) on gill histology and osmoregulation components of the gray mullet, Mugil cephalus

The gray mullet, Mugil cephalus is an inshore and bottom-feeding fish species of Oman sea. Therefore, the gray mullet may be more exposed to heavy metal contamination, as the toxic impacts of heavy metals mullet has been reported in various studies. This study was conducted to evaluate the toxic effects of the heavy metal, nickel (as NiCl2) on osmoregulation of the gray mullet by measuring blood biochemicals, hormones, minerals and gill histology. Fish (10 fish/tank) were experimentally exposed to NiCl2 at three environmentally relevant concentrations of 5, 10 and 15 μg/l for 96 h. Then, fish were challenged with seawater (35 mg/l) for a period of 120 min. The samples (blood and gill tissue) were collected After 96 exposure to NiCl2 and during salinity challenge (30, 60 and 120 min post challenge). The plasma levels of cortisol and glucose significantly increased in NiCl2−exposed fish. In addition, cortisol increased in all experimental groups 30 min after salinity challenge and then returned gradually to the same levels as the control at 120 min post salinity challenge (PSC). The triiodothyronine (T3) and thyroxine (T4) levels significantly decreased in response to 10 and 15 μg/l NiCl2. In all groups, the thyroid hormones significantly elevated at 30 min PSC. After 30 min PSC, T3 levels in all NiCl2−exposed fish and T4 in the treatment, 10 μg/l NiCl2 remained unchanged throughout the salinity challenge. In the treatment, 5 μg/l NiCl2, T4 levels were recovered at 120 min PSC and reached the same levels as the control. Exposure of fish to high concentrations of NiCl2 and salinity stress increased the lactate levels. However, lactate levels in 5 and 10 μg/l NiCl2 groups were recovered at 120 min PSC and reached the same levels as the control. Furthermore, plasma protein increased in response to 10 and 15 μg/l NiCl2. At 30 PSC, the protein levels decreased in control and 5 μg/l NiCl2 group, while it remained unchanged in fish exposed to 10 and 15 μg/l NiCl2 throughout the salinity challenge. Exposure of fish to NiCl2 disrupted the electrolyte (Na+, Cl−) balance both before and after salinity challenge, which may be due to gill lesions induced by the heavy metal and following alternations in gill permeability. However, fish in 5 μg/l NiCl2 re-established the ionic balance in the blood at the end of salinity challenge period. The malondialdehyde (MDA) levels significantly increased in response to 10 and 15 μg/l NiCl2. The MDA levels returned to the same levels as the control group at 120 min PSC. The results of the present study showed that nickel-induced toxicity (especially at high concentrations) can reduce the osmoregulation capabilities of mullet. However, fish are able to recover from the toxic effects over time, if contamination be eliminated.

CHANGES IN THE PLASMA ELECTROLYTES OF FRESH WATER TELEOST FISH LABEO ROHITA EXPOSED TO SUBLETHAL CONCENTRATION OF MONOSODIUM GLUTAMATE (MSG)

Background: Extensive usage of MSG in numerous food products as taste enhancer, and continuous release of efuents from Feed additive industries pose a threat to aquatic organisms.MSG induced cellular damages were reported in laboratory animals but there were no reports of MSG induced stress to shes. Methods: Fish Labeo rohita exposed to sublethal concentration of MSG (150mg/L) for 15 days and plasma electrolytes level were estimated. Results: MSG is toxic to sh even in sub lethal concentration. Decrease in plasma electrolytes during initial exposure to MSG may be due to + + disturbances in the membrane permeability of gills or inhibition of Na K ATP-ase activity in the gills. Elevated plasma electrolytes in MSG exposed sh may be due to efux of ions from their tissues there by indicating severe imbalance in the osmoregulation phenomena. Conclusion: MSG is toxic to sh and caused severe osmotic imbalance. Prolonged consumption of MSG through food as well as MSG intoxicated sh may affect human health via ecological cycling and biological magnication and cause serious health hazards like susceptibility to infectious diseases and even cancer

Interactive effects of temperature and hypoxia on diffusive water flux and oxygen uptake rate in the tidepool sculpin, Oligocottus maculosus

The osmorespiratory compromise hypothesis posits that respiratory epithelial characteristics and physiological regulatory mechanisms which promote gas permeability also increase permeability to ions and water. The hypothesis therefore predicts that physiological responses which increase effective gas permeability will result in increased effective ion and water permeabilities. Though analyses of water and gas effective permeabilities using high temperature have generally supported the hypothesis, water permeability responses to hypoxia remain equivocal and the combination of high temperature and hypoxia untested. We measured diffusive water flux (DWF) and oxygen uptake rate (Ṁo2) in response to acute temperature change, hypoxia, and the combination of high temperature and hypoxia in a hypoxia-tolerant intertidal fish, the tidepool sculpin (Oligocottus maculosus). In support of the osmorespiratory compromise hypothesis, Ṁo2 and DWF increased with temperature. In contrast, DWF decreased with hypoxia at a constant temperature, a result consistent with previously observed decoupling of water and gas effective permeabilities during hypoxia exposure in some hypoxia tolerant fishes. However, DWF levels during simultaneous high temperature and hypoxia exposure were not different from fish exposed to high temperature in normoxia, possibly suggesting a failure of the mechanism responsible for down-regulating DWF in hypoxia. These results, together with time-course analysis of hypoxia exposure and normoxic recovery, suggest that tidepool sculpins actively downregulate effective water permeability in hypoxia but the mechanism fails with multi-stressor exposure. Future investigations of the mechanistic basis of the regulation of gill permeability will be key to understanding the role of this regulatory ability in the persistence of this species in the dynamic intertidal environment.

Effects of repeated short episodes of environmental acidification on Atlantic salmon (Salmo salar) from a landlocked population

Chronic or repeated exposure to environmental contaminants may result in allostatic overload, a physiological situation in which the costs of coping affect long-term survival and reproductive output. Continuous measurements in Otra, the largest river in southern Norway, show the occurrence of repeated 24–48 h episodes of acidification. This work investigates the impact of repeated short acidification episodes on a unique land-locked population of normally anadromous Atlantic salmon (“Bleke”). This was done by recording physiological measures of stress and allostatic load in fish exposed for 7 days to continuous or repeated episodes of simulated environmental acidification or untreated Otra water (controls). A standardized acute stress test was performed after these different exposure regimes, with brain and blood samples taken before (baseline) or after the stress test. Treatment effects on stress coping ability were assessed by neuroendocrine indicators, including telencephalic serotonergic activity and plasma cortisol. Continuous exposure to acidification resulted in increased baseline plasma Cl− and Na+ and elevated baseline plasma cortisol compared to episodic exposed fish. However, both episodic and continuous acidification resulted in similar increase in gill Al, indicating similar impact on gill permeability of these two exposures. This suggests a lower impact on the electrolyte homeostasis in episodic compared to continuous exposure and that this effect is not directly related to the effects of Al complexes binding to the gills. Furthermore, there were no treatment induced differences on stress coping ability, suggesting that episodic exposure to the sublethal concentrations of Al in pH 5.5 in the present study do not result in higher allostatic load than in control or continuous exposed Bleke.

Ionoregulatory aspects of the hypoxia-induced osmorespiratory compromise in the euryhaline Atlantic killifish (Fundulus heteroclitus): the effects of salinity.

The osmorespiratory compromise is a physiological trade-off between the characteristics of the gill that promote respiratory gas exchange and those that limit passive flux of ions and water with the environment. In hypoxia, changes in gill blood flow patterns and functional surface area that increase gas transfer can promote an exacerbation in ion and water flux. Our goal was to determine whether the osmorespiratory compromise is flexible, depending on environmental salinity (fresh, isosmotic and sea water) and oxygen levels (hypoxia) in euryhaline killifish, Fundulus heteroclitus Plasma ion concentrations were minimally affected by hypoxia, indicating a maintenance of osmoregulatory homeostasis. In freshwater killifish, hypoxia exposure reduced branchial Na+/K+-ATPase and NEM-sensitive ATPase activities, as well as diffusive water flux rates. Unidirectional Na+ influx and Na+ efflux decreased during hypoxia in freshwater, but net Na+ flux remained unchanged. Net loss rates of Cl-, K+ and ammonia were also attenuated in hypoxia, suggesting both transcellular and paracellular reductions in permeability. These reductions appeared to be regulated phenomena as fluxes were restored immediately in normoxia. Na+ flux rates increased during hypoxia in 11 ppt, but decreased in 35 ppt, the latter suggesting a similar response to hypoxia to that in freshwater. In summary, freshwater and seawater killifish experience a reduction in gill permeability, as seen in other hypoxia-tolerant species. Fish acclimated to isosmotic salinity increased Na+ influx and efflux rates, as well as paracellular permeability in hypoxia, responses in accord with the predictions of the classic osmorespiratory compromise.

Plasma osmolality and oxygen consumption of perch Perca fluviatilis in response to different salinities and temperatures

The present study determined the blood plasma osmolality and oxygen consumption of the perch Perca fluviatilis at different salinities (0, 10 and 15) and temperatures (5, 10 and 20° C). Blood plasma osmolality increased with salinity at all temperatures. Standard metabolic rate (SMR) increased with salinity at 10 and 20° C. Maximum metabolic rate (MMR) and aerobic scope was lowest at salinity of 15 at 5° C, yet at 20° C, they were lowest at a salinity of 0. A cost of osmoregulation (SMR at a salinity of 0 and 15 compared with SMR at a salinity of 10) could only be detected at a salinity of 15 at 20° C, where it was 28%. The results show that P. fluviatilis have capacity to osmoregulate in hyper-osmotic environments. This contradicts previous studies and indicates intraspecific variability in osmoregulatory capabilities among P. fluviatilis populations or habitat origins. An apparent cost of osmoregulation (28%) at a salinity of 15 at 20° C indicates that the cost of osmoregulation in P. fluviatilis increases with temperature under hyperosmotic conditions and a power analysis showed that the cost of osmoregulation could be lower than 12·5% under other environmental conditions. The effect of salinity on MMR is possibly due to a reduction in gill permeability, initiated to reduce osmotic stress. An interaction between salinity and temperature on aerobic scope shows that high salinity habitats are energetically beneficial during warm periods (summer), whereas low salinity habitats are energetically beneficial during cold periods (winter). It is suggested, therefore, that the seasonal migrations of P. fluviatilis between brackish and fresh water is to select an environment that is optimal for metabolism and aerobic scope.

Gill paracellular permeability and the osmorespiratory compromise during exercise in the hypoxia-tolerant Amazonian oscar (Astronotus ocellatus)

In the traditional osmorespiratory compromise, fish increase their effective gill permeability to O2 during exercise or hypoxia, and in consequence suffer unfavorable ionic and osmotic fluxes. However oscars, which live in the frequently hypoxic ion-poor waters of the Amazon, actually decrease ionic fluxes across the gills during acute hypoxia without changing gill paracellular permeability, and exhibit rapid paving over of the mitochondrial-rich cells (MRCs). But what happens during prolonged exercise? Gill paracellular permeability, ionic fluxes, and gill morphology were examined in juvenile oscars at rest and during aerobic swimming. Initial validation tests with urinary catheterized fish quantified drinking, glomerular filtration, and urinary flow rates, and confirmed that measurements of gill paracellular permeability as [(3)H]PEG-4000 clearances were the same in efflux and influx directions, but far lower than previously measured in comparably sized trout. Although the oscars achieved a very similar proportional increase (90%) in oxygen consumption (MO2) to trout during steady-state swimming at 1.2 body lengths sec(-1), there was no increase in gill paracellular permeability, in contrast to trout. However, oscars did exhibit increased unidirectional Na(+) efflux and net K(+) rates during exercise, but no change in drinking rate. There were no changes in MRC numbers or exposure, or other alterations in gill morphology during exercise. A substantial interlamellar cell mass (ILCM) that covered the lamellae to a depth of 30% was unchanged by 4 h of swimming activity. We conclude that a low branchial paracellular permeability which can be dissociated from changes in O2 flux, as well as the presence of the ILCM, may be adaptive in limiting ionoregulatory costs for a species endemic to ion-poor, frequently hypoxic waters.

Enhanced Aeromonas salmonicida bacterin uptake and side effects caused by low frequency sonophoresis in rainbow trout (Oncorhynchus mykiss)

Low frequency sonophoresis (LFS) has been recognized as one of the most advanced technologies in transdermal delivery of substances, due to the modification of the stratum corneum lipid bilayer, in focal skin applications in mammals. Based on these findings, LFS has been suggested as a potential technology to be used for enhancement in immersion fish vaccination. In contrast to mammals where LFS is applied to discrete regions of the skin, in fish the whole individual needs to be exposed for practical purposes. The current study evaluated the impact of LFS at 37 kHz on the uptake of an Aeromonas salmonicida bacterin and side effects of the treatment in rainbow trout. Quantitative real time PCR (qPCR) and immunohistochemistry were used to examine the bacterin uptake into skin and gill tissue. Side effects were assessed by behavioural examination, histology and blood serum analysis. The sonication intensity of 171 mW/cm2 was enough for increasing skin permeability, but caused heavy erratic swimming and gill haemorrhages. Sonication intensities as low as 105 mW/cm2 did not modify skin permeability and enhanced the bacterin uptake into the gill tissue by factor 15 compared to conventional immersion. Following sonication, the gill permeability for the bacterin decreased after 20 min and 120 min by factor 3 and 2, respectively. However, during sonication, erratic swimming of the fish raised some concerns. Further reduction of the sonication intensity to 57 mW/cm2 did not induce erratic swimming, and the bacterin uptake into the gill tissue was still increased by factor 3. In addition, a decreasing albumin–globulin ratio in the serum of the rainbow trout within 40 min revealed that LFS leads to an inflammatory response. Consequently, based on both increased bacterin uptake and the inflammatory response, low intensity LFS has the potential to enhance vaccine immunity without significant side effects.

Progress toward understanding the bioaccumulation of perfluorinated alkyl acids

Progress toward understanding the bioaccumulation of perfluorinated alkyl acids

Differential responses in ammonia excretion, sodium fluxes and gill permeability explain different sensitivities to acute high environmental ammonia in three freshwater teleosts

We examined the acute physiological responses to high environmental ammonia (HEA), particularly the linkages between branchial ammonia fluxes and unidirectional Na+ fluxes, as well as urea excretion, cortisol, and indicators of gill permeability in three freshwater teleosts differing in their sensitivities to ammonia; the highly sensitive salmonid Oncorhynchus mykiss (rainbow trout), the less sensitive cyprinid Cyprinus carpio (common carp) and the highly resistant cyprinid Carassius auratus (goldfish). Fish were acutely exposed to two sub-lethal ammonia concentrations (as NH4HCO3) at pH 7.9: 1mM for a period of 12h, identical for all species, and 5mM for the cyprinids and 1.4mM for the trout for 3h. Elevation of plasma cortisol at both levels of HEA was apparent in all species. At 1mM, ammonia excretion (Jamm) was inhibited to a greater extent in trout than cyprinids and concurrently a significantly higher plasma ammonia level was evident in trout. However Jamm was reversed in all species at 5 or 1.4mM. Goldfish showed a significant increase in urea excretion rate (Jurea) during HEA exposure. In carp and trout, neither level of HEA elevated Jurea but urea production was increased as evidenced by a considerable elevation of plasma urea. At 1mM HEA, Na+ imbalance became progressively more severe in trout and carp due to a stimulation of unidirectional Na+ efflux (JoutNa) without a concomitant increase in unidirectional Na+ influx (JinNa). Additionally, a transient reduction of JinNa was evident in trout. Goldfish showed an opposite trend for JoutNa with reduced efflux rates and a positive Na+ balance during the first few hours of HEA. However, after 12h of exposure, both JinNa and JoutNa were also increased in both carp and goldfish, whereas only JoutNa was increased in trout, leading to a net Na+ loss. Na+ homeostasis was entirely disrupted in all three species when subjected to the 5 or 1.4mM ammonia for 3h: JinNa was significantly inhibited while considerable activation of JoutNa was observed. Diffusive water efflux rates and net K+ loss rates across the gills were enhanced during HEA only in trout, indicating an increment in gill transcellular permeability. Transepithelial potential was increased in all the species during ammonia exposure, but to the least extent in goldfish. Overall, for several different physiological systems, trout were most disturbed, and goldfish were least disturbed by HEA, helping to explain the differential ammonia tolerance of the three species.

Permeability properties of the teleost gill epithelium under ion-poor conditions

Permeability properties of the goldfish gill epithelium were examined in vivo and in vitro following exposure to ion-poor water (IPW) conditions. In gill tissue of IPW-acclimated goldfish, transcript abundance of tight junction (TJ) proteins occludin, claudin-b, -d, -e, -h, -7, and -8d increased, whereas ZO-1 and claudin 12 mRNA decreased and claudin-c was unaltered. In association with these changes, TJ depth increased among gill pavement cells (PVCs) and gill PVCs and mitochondria-rich cells (MRCs). PVC and MRC gill cell fractions were isolated using Percoll. Transcripts encoding for occludin, claudin-b, -c, -d, -e, -h, -7, -8d, -12, and ZO-1 were present in both fractions. After IPW acclimation, occludin, claudin-b and -e, and ZO-1 mRNA abundance increased in both fractions. In contrast, claudin-8d mRNA abundance increased in PVCs only while claudin-h decreased in MRCs. Gill permeability was examined using primary cultured goldfish PVC epithelia supplemented with serum derived from IPW-acclimated goldfish. IPW serum supplementation increased transepithelial resistance, reduced [(3)H]PEG-4000 permeability, and enhanced epithelial integrity during in vitro IPW exposure. IPW serum increased mRNA abundance of occludin, claudin-8d and -e in vitro. Using small interfering RNA, we found that occludin abundance was decreased in cultured gill epithelia, resulting in an increase in [(3)H]PEG-4000 flux. As occludin increased in the gills of IPW-acclimated fish as well as cultured gill epithelia exposed to IPW serum, results suggest that occludin is a barrier-forming TJ protein in fish gill epithelia. These studies support the idea that TJ proteins play an important role in regulating gill permeability in IPW.

Glucocorticoid and mineralocorticoid receptors regulate paracellular permeability in a primary cultured gill epithelium

The role of corticosteroid receptors (CRs) in the regulation of gill permeability was examined using a primary cultured trout gill epithelium. The epithelium expressed both glucocorticoid receptors (GR1 and GR2) and a mineralocorticoid receptor (MR), and cortisol treatment significantly increased transepithelial resistance (TER) and decreased paracellular [(3)H]PEG-4000 flux. Epithelial permeability was unaffected by deoxycorticosterone or aldosterone. The GR antagonist RU486 as well as MR antagonists spironolactone and RU26752 significantly reduced, but did not completely block, the effects of cortisol. The MR antagonist eplerenone was without effect. Only RU486 + spironolactone or RU486 + RU26752 treatment completely suppressed the effects of cortisol. On its own, RU486 had cortisol-like effects which could be blocked by spironolactone, suggesting that although RU486 is a GR antagonist, in this system it may also have agonistic properties that are mediated through the MR. The GR agonist dexamethasone increased TER and reduced [(3)H]PEG-4000 flux across cultured epithelia and was unaffected by MR antagonists. Therefore, alterations in transcript abundance of select tight junction (TJ) proteins were examined in response to cortisol, dexamethasone (a GR agonist) and RU486 (as a MR agonist). Occludin and claudin-7, -8d, -12 and -31 mRNA were significantly elevated in response to cortisol, dexamethasone or RU486 treatment. Claudin-3a mRNA was significantly elevated in response to cortisol or dexamethasone only, and claudin-28b and -30 mRNA were significantly altered following cortisol or RU486 treatment only. The data support a role for the GRs and MR in regulating gill permeability and suggest that TJ proteins are responsive to cortisol through both or individual CR types.

Permeability properties and occludin expression in a primary cultured model gill epithelium from the stenohaline freshwater goldfish

Techniques for the primary culture of fish gill epithelia on permeable supports have provided 'reconstructed' gill models appropriate for the study of gill permeability characteristics in vitro. Models developed thus far have been derived from euryhaline fish species that can tolerate a wide range of environmental salinity. This study reports on procedures for the primary culture of a model gill epithelium derived from goldfish, a stenohaline freshwater (FW) fish that cannot tolerate high environmental salt concentrations. The reconstructed goldfish gill epithelium was cultured on permeable filter inserts and using electron microscopy and immunocytochemical techniques, was determined to be composed exclusively of gill pavement cells. When cultured under symmetrical conditions (i.e. with culture medium bathing both apical and basolateral surfaces), epithelial preparations generated appreciable transepithelial resistance (TER) (e.g. 1,150 ± 46 Ωcm(2)) within 36-42 h post-seeding in inserts. When apical medium was replaced with FW (asymmetrical conditions to mimic conditions that occur in vivo), epithelia exhibited increased TER and elevated paracellular permeability. Changes in permeability occurred in association with altered occludin-immunoreactive band position by western blot and no change in occludin mRNA abundance. We contend that the goldfish gill model will provide a useful in vitro tool for examining the molecular components of a stenohaline fish gill epithelium that participate in the regulation of gill permeability. The model will allow molecular observations to be made together with assessment of changing physiological properties that relate to permeability. Together, this will allow further insight into mechanisms that regulate gill permeability in fishes.

A Simple Allometric Diffusion-Based Biokinetic Model to Predict Cu(II) Uptake Across Gills of Freshwater Clam Corbicula fluminea

The purpose of this study was to link Fick's type mass transfer and biokinetics together with Michaelis-Menten kinetics to arrive at a simple predictive framework for quantifying biouptake mechanisms in gills of freshwater clam Corbicula fluminea exposed to Cu(II). A diffusion-based Cu(II) influx and permeability can be calculated using physiological and allometric-related parameters. Simulations indicate that Cu(II) bioconcentration factor of gills was 42. Estimated steady-state Cu(II) gill uptake influx and permeability were 0.097 nmol cm(-2) s(-1) and 0.48 cm s(-1), respectively. The proposed simple allometric diffusion-based biokinetic model meets the need for describing nonequilibrium aspects of biouptake mechanisms in bivalve gills.

Cortisol differentially alters claudin isoforms in cultured puffer fish gill epithelia

A primary cultured gill epithelium from the puffer fish Tetraodon nigroviridis was developed to examine the corticosteroid regulation of claudin isoform mRNA abundance in fish gills. Preparations were composed of polygonal epithelial cells exhibiting concentric apical microridges and zonula occludens-1 immunoreactivity along cell margins. No evidence was found to indicate the presence of Na +–K +–ATPase-immunoreactive or mitochondria-rich cells in cultured preparations. Therefore, epithelia appear to be composed of gill pavement cells (PVCs) only. An RT-PCR profile of 12 salinity responsive gill claudin tight junction (TJ) proteins ( Tncldn3a, - 3c, - 6, - 8d, - 10d, - 10e, - 11a, - 23b, - 27a, - 27c, - 32a, and - 33b) revealed the absence of Tncldn6, - 10d and - 10e in cultured epithelia, suggesting that these isoforms are not associated with gill PVCs. Cortisol treatment of cultured epithelia dose-dependently increased or decreased mRNA abundance of select claudin isoforms. Transcript abundance of several claudin isoforms was unaffected by cortisol treatment. These data provide evidence for the cell specific distribution of claudins in fish gills and suggest that heterogeneous alterations in the abundance of select claudin isoforms contribute to the corticosteroid regulation of gill permeability.

Effect of hypo- and hypersaline conditions on osmolality and Na+/K+-ATPase activity in juvenile shrimp (Litopenaeus vannamei) fed low- and high-HUFA diets

Fatty acid composition of cellular membranes can modify permeability and can modulate the activity of Na(+)/K(+)-ATPase. Although highly unsaturated fatty acids (HUFA) improve survival and osmoregulatory capacity to low salinities in penaeid shrimp, the possible mechanisms have not been established. For this purpose the influence of HUFA supplementation in diet (2.9 vs. 34% HUFA proportion to total fatty acids) on osmoregulatory responses of juvenile Litopenaeus vannamei submitted to an acute (15 h) or chronic exposure (21 days), to low (5 g L(-1)) and high salinities (50 g L(-1)) was analyzed. Shrimp fed the high-HUFA diet, had higher concentration of main HUFA (20:5n-3 and 22:6n-3) in polar lipids of gills. Osmotic pressure in hemolymph was significantly affected by salinity in acute (640, 751, 847 mOsm/kg for 5, 30 and 50 g L(-1), respectively), and chronic exposure (645, 713, 814 mOsm/kg), but variations between them were small compared to environmental salinity (206, 832, 1547 mOsm/kg), indicating that osmoregulation was achieved in a matter of hours. An increase in Na(+)/K(+)-ATPase activity was observed only after a chronic exposure to low salinity. Free amino acids (FAA), mainly alanine and arginine, were higher at 30 (control) and 50 g L(-1) in accordance to their role as organic osmolites. Neither osmotic pressure, Na(+)/K(+)-ATPase activity, nor FAA was affected by HUFA supplementation. However, higher water content in gills of shrimp exposed to low salinities was counteracted by increased HUFA content, which could be a result of changes in water permeability of gills. The osmoregulatory capacity of penaeid shrimp to low and high salinities was achieved within 15 h of acclimation and did not depend on HUFA supplementation in the diet.

Gill area, permeability and Na+,K+-ATPase activity as a function of size and salinity in the blue crab,Callinectes sapidus

Juvenile blue crabs, Callinectes sapidus, extensively utilize oligohaline and freshwater regions of the estuary. With a presumptively larger surface-area-to-body weight ratio, juvenile crabs could experience osmo- and ionoregulatory costs well in excess of that of adults. To test this hypothesis, crabs ranging over three orders of magnitude in body weight were acclimated to either sea water (1,000 mOsm) or dilute sea water (150 mOsm), and gill surface area, water and sodium permeabilities (calculated from the passive efflux of 3H2O and 22Na+), gill Na+, K+ -ATPase activity and expression were measured. Juveniles had a relatively larger gill surface area; weight-specific gill surface area decreased with body weight. Weight-specific water and sodium fluxes also decreased with weight, but not to the same extent as gill surface area; thus juveniles were able to decrease gill permeability slightly more than adults upon acclimation to dilute media. Crabs < 5 g in body weight had markedly higher activities of gill Na+ ,K+ -ATPase than crabs > 5 g in both posterior and anterior gills. Acclimation to dilute medium induced increased expression of Na+, K+ -ATPase and enzyme activity, but the increase was not as great in juveniles as in larger crabs. The increased weight-specific surface area for water gain and salt loss for small crabs in dilute media presents a challenge that is incompletely compensated by reduced permeability and increased affinity of gill Na+, K+ -ATPase for Na+. Juveniles maintain osmotic and ionic homeostasis by the expression and utilization of extremely high levels of gill Na+, K+ -ATPase, in posterior, as well as in anterior, gills.

Diminished social status affects ionoregulation at the gills and kidney in rainbow trout (Oncorhynchus mykiss)

Competition for social status can result in physiological differences between individuals, including differences in ionoregulatory ability. Subordinate rainbow trout (Oncorhynchus mykiss) had two-fold higher uptake rates of sodium across the gill and two-fold higher whole-body sodium efflux rates than the dominant fish with which they were paired. Sodium efflux was then divided into branchial and renal components, both of which were higher in subordinates. Branchial sodium efflux accounted for 95%–98% of sodium loss. Plasma sodium concentrations were more variable, although not significantly different, in subordinate fish, suggesting that the increased loss of sodium in these trout is compensated for by an increase in uptake rates. Urine flow rates and plasma cortisol concentrations were higher in subordinate fish, but there was no difference in glomerular filtration rate between dominants and subordinates. Renal sodium reabsorption was significantly reduced in subordinates. In summary, the ionoregulation of subordinate individuals was altered, most likely occurring as a result of stress-induced changes in gill permeability, resulting in a higher throughput of water and increased branchial sodium efflux. These changes in ionoregulatory ability have many physiological implications, including the increased susceptibility of subordinates to ionoregulatory challenges and an increased metabolic cost of ionoregulation.

Bioconcentration and tissue distribution of perfluorinated acids in rainbow trout (Oncorhynchus mykiss)

Rainbow trout (Oncorhynchus mykiss) were exposed simultaneously to a homologous series of perfluoroalkyl carboxylates and sulfonates in a flow-through system to determine compound-specific tissue distribution and bioconcentration parameters for perfluorinated acids (PFAs). In general, PFAs accumulated to the greatest extent in blood > kidney > liver > gall bladder. Carboxylates and sulfonates with perfluoroalkyl chain lengths shorter than seven and six carbons, respectively, could not be detected in most tissues and were considered to have insignificant bioconcentration factors (BCFs). For detectable PFAs, carcass BCFs increased with increasing length of the perfluoroalkyl chain, ranging from 4.0 to 23,000, based on wet weight concentrations. Carboxylate carcass BCFs increased by a factor of eight for each additional carbon in the perfluoroalkyl chain between 8 and 12 carbons, but this relationship deviated from linearity for the longest PFA tested, possibly because of decreased gill permeability. In general, half-lives (3.9-28 d) and uptake rates (0.053-1.700 L/kg/d) also increased with increasing length of the perfluoroalkyl chain in all tissues. Sulfonates had greater BCFs, half-lives, and rates of uptake than the corresponding carboxylate of equal perfluoroalkyl chain length, indicating that hydrophobicity, as predicted by the critical micelle concentration, is not the only determinant of PFA bioaccumulation potential and that the acid function must be considered.

Ammonia excretion and urea handling by fish gills: present understanding and future research challenges.

In fresh water fishes, ammonia is excreted across the branchial epithelium via passive NH(3) diffusion. This NH(3) is subsequently trapped as NH(4)(+) in an acidic unstirred boundary layer lying next to the gill, which maintains the blood-to-gill water NH(3) partial pressure gradient. Whole animal, in situ, ultrastructural and molecular approaches suggest that boundary layer acidification results from the hydration of CO(2) in the expired gill water, and to a lesser extent H(+) excretion mediated by apical H(+)-ATPases. Boundary layer acidification is insignificant in highly buffered sea water, where ammonia excretion proceeds via NH(3) diffusion, as well as passive NH(4)(+) diffusion due to the greater ionic permeability of marine fish gills. Although Na(+)/H(+) exchangers (NHE) have been isolated in marine fish gills, possible Na(+)/NH(4)(+) exchange via these proteins awaits evaluation using modern electrophysiological and molecular techniques. Although urea excretion (J(Urea)) was thought to be via passive diffusion, it is now clear that branchial urea handling requires specialized urea transporters. Four urea transporters have been cloned in fishes, including the shark kidney urea transporter (shUT), which is a facilitated urea transporter similar to the mammalian renal UT-A2 transporter. Another urea transporter, characterized but not yet cloned, is the basolateral, Na(+) dependent urea antiporter of the dogfish gill, which is essential for urea retention in ureosmotic elasmobranchs. In ureotelic teleosts such as the Lake Magadi tilapia and the gulf toadfish, the cloned mtUT and tUT are facilitated urea transporters involved in J(Urea). A basolateral urea transporter recently cloned from the gill of the Japanese eel (eUT) may actually be important for urea retention during salt water acclimation. A multi-faceted approach, incorporating whole animal, histological, biochemical, pharmacological, and molecular techniques is required to learn more about the location, mechanism of action, and functional significance of urea transporters in fishes.