Branchial and intestinal osmoregulatory acclimation in the four-eyed sleeper, <Emphasis Type="Italic">Bostrychus sinensis</Emphasis> (Lacepède), exposed to seawater

Bostrychus sinensis is a facultative air breather that inhabits waters of a wide range of salinities. This study aimed to elucidate whether branchial and intestinal osmoregulatory acclimation occurred in B. sinensis transferred from 5‰ water through a progressive increase in salinities to seawater. Our results indicate that B. sinensis acted as a hyperosmotic regulator in 5‰ water, but exhibited hypoosmotic hypoionic regulation in seawater. During short- (1 day) and medium- (10 days) term acclimation to seawater, there were only minor perturbations in plasma osmolality and [Na⁺], which returned to control levels after 45 days of exposure to seawater. Branchial Na⁺/K⁺-ATPase activity was unaffected by 1, 10 or 45 days of exposure to seawater. However, prolonged (45 days) acclimation to seawater led to a significant increase in Na⁺/K⁺-ATPase α-subunit protein abundance. Taken together, these results indicate that there could be changes in the expression of Na⁺/K⁺-ATPase isoforms and/or post-translational modification of Na⁺/K⁺-ATPase in the gills of fish exposed to seawater. Immunofluorescence microscopy revealed that acclimation to seawater for 10 days only resulted in no change in branchial Na⁺/K⁺-ATPase protein expression, but there were increases in protein expression of cystic fibrosis transmembrane regulator (CFTR)-like chloride channel and Na⁺:K⁺:2Cl⁻ cotransporter (NKCC; probably NKCC1). Indeed, NKCC was undetectable in gills of fish kept in 5‰ water by Western blotting, but it became weakly detectable in fish exposed to seawater for 10 days and prominently expressed in fish exposed to seawater for 45 days. Therefore, our results indicate that branchial CFTR-like chloride channel and NKCC1 were the determining factors in the transition between hyperosmotic regulation and hypoosmotic hypoionic regulation in B. sinensis. Furthermore, the intestine of B. sinensis also served as an important osmoregulatory organ, since there were significant increases in both the activity and protein abundance of intestinal Na⁺/K⁺-ATPase in fish acclimated to seawater for 45 days. The effectiveness of branchial and intestinal osmoregulatory acclimation in B. sinensis during seawater acclimation led to only a minor increase in plasma osmolality, and thus resulted in relatively unchanged free amino acid contents in muscle and liver.     

Interaction of short-term testosterone treatment with osmotic acclimation in the gilthead sea bream <Emphasis Type="Italic">Sparus auratus</Emphasis>

To assess the interaction between testosterone (T) treatment and acclimation to different salinities, seawater-acclimated gilthead sea bream (Sparus auratus) were implanted with slow-release coconut oil implants alone (control) or containing T (5 μg/g body mass). After 5 days, eight fish of control and T-treated groups were sampled. The same day, eight fish of each group were transferred to low salinity water (LSW, 6 ppt, hypoosmotic test), seawater (SW, 38 ppt, control test) and high salinity water (HSW, 55 ppt, hyperosmotic test) and sampled 9 days later. Gill Na⁺, K⁺-ATPase activity increased in HSW-acclimated fish with respect to SW- and LSW-acclimated fish in both control and T-treated groups. Kidney Na⁺, K⁺-ATPase activity was also enhanced in HSW-acclimated fish, but only in T-treated group. From a metabolic point of view, most of the changes observed can be attributed to the action of salinity and T treatment alone, since few interactions between T treatment and osmotic acclimation to different salinities were observed. Those interactions included in treated fish: in the liver, decreased capacity in using glucose in fish acclimated to extreme salinities; in the gills, decreased capacity in using amino acids in HSW; in the kidneys increased capacity in using amino acids in extreme salinities; and in the brain, decreased glycogen and acetoacetate levels of fish in LSW.     

Acclimation of Solea senegalensis to different ambient temperatures: implications for thyroidal status and osmoregulation

We have investigated the regulation of thyroidal status and osmoregulatory capacities in juveniles from the teleost Solea senegalensis acclimated to different ambient temperatures. Juveniles, raised in seawater at 19°C, were acclimated for 3 weeks to temperatures of 12, 19 and 26°C. Since our preliminary observations showed that at 12°C feed intake was suppressed, our experimental design controlled for this factor. The concentration of branchial Na⁺,K⁺-ATPase, estimated by measurements of enzyme activity at the optimum temperature of this enzyme (37°C), did not change. In contrast, an increase in Na⁺,K⁺-ATPase activity (measured at 37°C), was observed in the kidney of 12°C-acclimated fish. In fish acclimated to 12°C, the hepatosomatic index had increased, which correlated with increased plasma levels of triglycerides and non-esterified fatty acids. Plasma cortisol levels did not differ significantly between the experimental groups. In liver and gills, the amount of iodothyronine deiodinases that exhibit thyroid hormone outer ring deiodination was up-regulated only when fish did not feed. When assayed at the acclimation temperature, kidney deiodinase activities were similar, indicating a temperature-compensation strategy. 3,5,3′-triiodothyronine (T3) tissue concentrations in gills and kidney did not differ significantly between experimental groups. However, at 12°C, lower T3 tissue levels were measured in plasma and liver. We conclude that S. senegalensis adjusts its osmoregulatory system to compensate for the effects of temperature on electrolyte transport capacity. The organ-specific changes in thyroid hormone metabolism at different temperatures indicate the involvement of thyroid hormones in temperature acclimation.     

A comparison of transport-related gill enzyme activities and tissue-specific free amino acid concentrations of Baltic Sea (brackish water) and freshwater threespine sticklebacks, Gasterosteus aculeatus, after salinity and temperature acclimation

The osmoregulatory abilities of one freshwater and two brackish water (Baltic Sea) populations of the euryhaline teleost fish Gasterosteus aculeatus were studied with respect to evolutionary physiology. Plasma osmolality, activities of Na⁺K⁺-ATPase, citrate synthase, creatine kinase in the gill and free amino acids in liver, axial muscle and pectoral fin muscle were measured. After transfer from 10 to 35 ppt at 15 °C, time-course changes of plasma osmolality and gill Na⁺K⁺-ATPase showed no significant fundamental differences between the freshwater and one of the Baltic Sea populations. In a multi-factorial experiment, each population was exposed to four different abiotic regimes. Both brackish water populations had high mortality in freshwater at 4 °C, which is discussed as a failure of osmotic regulation (reduced taurine concentrations). Freshwater specimens had higher levels of glycine in the axial and pectoral fin muscles compared to the brackish water populations. This is interpreted as a genetically based effect. In brackish (20 ppt) water of 15 °C, the freshwater population had high activities of Na⁺K⁺-ATPase, but low activities of creatine kinase, whereas both brackish water populations behaved in the opposite way. A fundamental difference between the freshwater and brackish water populations on the level of the osmoregulatory machinery was not observed. Received: 10 December 1998 / Accepted: 22 September 1999     

Na+-K+-adenosine triphosphatase activities in gills of marine teleost fishes: Changes with depth,size and locomotory activity level

Activities of the primary enzyme responsible for monovalent ion regulation, Na⁺-K⁺-adenosine triphosphatase (Na⁺-K⁺-ATPase), were measured in gills of marine teleost fishes with different depths of occurrence (0 to 4800 m), body weights (a range of five orders of magnitude), and locomotory capacities. Specimens were collected off the coasts of California and Oregon in 1983–1989, and at the Galápagos Spreading Center and 13°N East Pacific Rise hydrothermal vent sites in 1987 and 1988, respectively. Except for two hydrothermal vent fishes, deep-sea species had much lower Na⁺-K⁺-ATPase activities g^–1 gill filament than shallow-living species, indicating that osmoregulatory costs, like total metabolic rate, are greatly reduced in most deep-living fishes. Within a species, the total branchial Na⁺-K⁺-ATPase activity per individual was dependent on size; the average allometric scaling exponent was 0.83. Using published values for oxygen consumption rates, and the total branchial Na⁺-K⁺-ATPase activities as an index of osmoregulatory costs, we estimated the maximal cost (as percent of ATP turnover) for osmoregulation in ten teleosts. Osmoregulatory costs averaged about 10% of total ATP turnover among these species, and maximal costs were no greater than about 20%. The percent costs of osmoregulation did not differ between shallow- and deep-living fishes. The reduced total ATP expenditure for osmoregulation in deep-living fishes is proposed to result from the sluggish locomotory habits of these fishes, not from selection for reduced osmotic coastper se. Thus, the reduced swimming abilities of these fishes lead to lower rates of water flow over the gills and less blood flow through the gills due to reduced demands for oxygen. Consequently, passive flux of water and ions through the gills is much lower than in more active fishes, and osmotic costs are thereby minimized. The relatively high activities of Na⁺-K⁺-ATPase in gills of the two hydrothermal vent fishes suggest that these fishes may be more active and have higher metabolic rates than other deep-sea fishes.     

Sodium- and potassium-activated adenosine triphosphatase in the excretory organs of Sepia officinalis (Cephalopoda)

Sodium- and potassium-activated ATPase (Na⁺–K⁺-ATPase) has been demonstrated in excretory organs of Sepia officinalis, using a cytochemical procedure. In the renal appendages, both epithelia of the pancreatic appendages, the folded epithelium of the branchial heart appendage and the transport-active epithelium of the gill, the enzyme is localized exclusively in the basolateral cell membranes, i.e., the membranes of the basal labyrinth and the lateral plasma membranes. In addition, Na⁺–K⁺-ATPase is also located in the sarcolemma of the muscle fibres of the branchial heart. Distribution and localization of the enzyme is further substantiated by [³H]-ouabain autoradiography. The possible involvement of Na⁺–K⁺-ATPase in the excretion of ammonia and in ionic regulation in dibranchiate cephalopods is discussed.This study was supported by the Deutsche Forschungsgemeinschaft and is part of a doctoral dissertation     

Reduced hypoosmoregulatory ability and alteration in gill chloride cell distribution in mature chum salmon (Oncorhynchus keta) migrating upstream for spawning

Osmoregulatory ability of mature chum salmon (Oncorhynchus keta) during spawning migration was examined by following the changes in gill Na⁺, K⁺-ATPase activity and in the distribution and morphology of chloride cells. Mature chum salmon caught in Otsuchi Bay, northern Honshu Island, Japan, died within 5 d in seawater (SW) in association with a marked increase in plasma osmolality, whereas the fish transferred to fresh water (FW) maintained plasma osmolality efficiently. Gill Na⁺, K⁺-ATPase activity decreased in both SW-maintained and FW-transferred fish. Well-developed chloride cells, identified by immunocytochemical staining specific for Na⁺, K⁺-ATPase, were present mainly in the filament epithelium of immature fish caught in the ocean. In mature fish caught in the bay, however, additional chloride cells were also found in the lamellar epithelium. The number of filament chloride cells decreased markedly in the mature fish both in SW and in FW, whereas the number of lamellar chloride cells was maintained. These results suggest that the loss of hypoosmoregulatory ability in mature chum salmon may be attributable to the decrease in filament chloride cells and associated decrease in gill Na⁺, K⁺-ATPase activity, and also that appearance of lamellar chloride cells may be preparatory to the forthcoming upstream migration. Received: 14 April 1997 / Accepted: 5 May 1997     

Ontogenic change of gill chloride cells in leptocephalus and glass eel stages of the Japanese eel, <Emphasis Type="Italic">Anguilla japonica</Emphasis>

The development of gill chloride cells was examined in premetamorphic larvae (leptocephali) and juveniles (glass eels) of the Japanese eel, Anguilla japonica. Branchial chloride cells were detected by immunocytochemistry using an antiserum specific for Na⁺,K⁺-ATPase. The specificity and availability of the antiserum for the detection of Japanese eel chloride cells were confirmed by Western blot analysis. The chloride cells first appeared on the developing gill filaments in a mid larval stage of leptocephalus (32.2 mm). Both immunoreactivity and the number of chloride cells gradually increased as the fish grew to a late stage of leptocephalus over 54 mm. In glass eels just after metamorphosis, gill lamellae developed from the gill filaments, and a rich population of chloride cells was observed in the gill filaments. In glass eels collected at a coastal area, chloride cells were extensively distributed in the gill filaments. The chloride cell size decreased progressively in glass eels transferred from seawater (SW) to freshwater (FW), whereas there was no difference in cell number. In contrast, some Na⁺,K⁺-ATPase immunoreaction distinct from typical chloride cells was observed in the gill lamellae throughout FW-transferred fish, but disappeared in control fish maintained in SW for 14 days. These findings indicate that the gill and gill chloride cells developed slowly during the extremely long larval stage, followed by rapid differentiation during a short period of metamorphosis. The excellent euryhalinity of glass eels may be due to the presence of the filament chloride cells and lamellar Na⁺,K⁺-ATPase-immunoreaction, presumably being responsible for SW and FW adaptation, respectively.     

Comparison of the osmoregulatory capabilities of two portunid crabs,Callinectes sapidus and C. similis

An investigation of the osmoregulatory capabilities of two portunid crabs, Callinectes sapidus and C. similis, was conducted to determine if their differences in distributional patterns were reflected in their capacity to adjust physiologically to changes in salinity. After acclimation to 5, 20 and 35 S, measurements of hemolymph and muscle concentrations of Na⁺, Cl^- and K⁺ and muscle-free amino acids indicated that C. sapidus is a better osmoregulator at low salinity than C. similis, while both species osmoregulate equally well at high salinity. This difference in osmoregulatory capacity corresponds well with their distribution in coastal-plain estuaries.This research was supported under agreement (49-7)-5 between the National Marine Fisheries Service and the Energy Research and Development Administration.Communicated by M.R. Tripp, Newark     

Water balance systems of Rangia cuneata: ionic and amino acid regulation in changing salinities

In order to study the interaction of the extracellular and intracellular osmoregulatory systems of the bivalve Rangia cuneata, we have measured blood osmotic and ionic concentrations together with intracellular free amino acid concentrations and total tissue water under identical salinity conditions. Like freshwater bivalves, the blood of R. cuneata is maintained hyperosmotic (50 mOsm) to the environment in salinities below 110 mosm by the regulation of Na⁺, Cl^-, K⁺ and Ca²⁺ concentrations. On the other hand in company with marine bivalves, R. cuneata also regulates intracellular free amino acids (FAA) as a mechanism to control cellular volume during osmotic stress over the entire non-lethal salinity range (3 to 620 mOsm). Alanine is the predominant intracellular osmotic effector. Thus, by utilizing the osmoregulatory mechanisms of both marine and freshwater bivalves, R. cuneata is able to tolerate salinities ranging from freshwater to 25 ppt and to traverse the faunal salinity boundary, known as the horohalinicum (5 to 8 ppt), controlling cell volume throughout.Please address requests for reprints to Dr. S. K. Pierce     

Inhibition of Na+/K+-ATPase and of active ion-transport functions in the gills of the shore crab Carcinus maenas induced by cadmium

Inhibition of Na⁺/K⁺-ATPase from gill plasma membranes of the shore crab Carcinus maenas by cadmium was investigated and compared with inhibitory effects by known antagonists (ouabain and Ca²⁺). For comparative considerations the Cd²⁺-inhibition of the enzyme from dog kidney was also tested. Na⁺/K⁺-ATPase from dog kidney and from crab gill differed greatly in sensitivity against ouabain. The inhibition constant K _i of the dog enzyme amounted to 9.1 × 10⁻⁷ mol l⁻¹, i.e. more than 300-fold smaller than the K _i of 2.9 × 10⁻⁴ mol l⁻¹ determined for the crab enzyme. Ca²⁺ inhibited the activity of Na⁺/K⁺-ATPase from crab gill plasma membranes with a K _i of 4.3 × 10⁻⁴ mol l⁻¹. The Na⁺/K⁺-ATPase from crab gill was inhibited by Cd²⁺ with a K _i of 9.1 × 10⁻⁵ mol l⁻¹. Cd²⁺ inhibited the Na⁺/K⁺-ATPase from dog kidney with a K _i (6.4 × 10⁻⁵ mol l⁻¹) comparable to that observed in the crab gill enzyme. Under experimental conditions Cd²⁺-inhibition of Na⁺/K⁺-ATPase was irreversible. Repeated washing, centrifugation and homogenization of the plasma membranes (four times) with Cd²⁺-free buffer did not restore any activity lost in the presence of 1 × 10⁻³ mol l⁻¹ Cd²⁺. Since ouabain-insensitive (nonspecific) ATPases in the plasma membrane fraction of crab gills were inhibited by Cd²⁺ in the same way as Na⁺/K⁺-ATPase, the heavy metal is considered as an unspecific ATPase inhibitor. Comparing these results with literature data on Cd²⁺-binding to electrophoretically separated proteins suggests that Na⁺/K⁺-ATPase is a Cd²⁺-binding enzyme. The results obtained on Na⁺/K⁺-ATPase were reflected by Cd²⁺-inhibition of the branchial ion-transport functions depending on this enzyme. The transepithelial short-circuit current of isolated gill half lamellae, a direct measure of area-specific active ion uptake, and the transepithelial potential difference of isolated, perfused whole gills, also indicative of active ion uptake, were inhibited by the heavy metal in a time- and dose-dependent mode. Remarkably these inhibitions were also irreversible. These findings are ecologically and biomedically significant: even when the actual environmental or tissue concentrations measured are low, biological microstructures such as Na⁺/K⁺-ATPase may accumulate the heavy metal by tight binding over prolonged periods until the first inhibitory effects occur. Received: 25 June 1997 / Accepted: 25 August 1997     

Evidence that the rotifer<Emphasis Type="Italic"> Brachionus plicatilis</Emphasis> is not an osmoconformer

The rotifer Brachionus plicatilis is euryhaline (growing between 2 and 97 ppt) and has previously been considered an osmoconformer. We suggest that B. plicatilis is an osmoregulator, exhibiting a pattern of Na⁺/K⁺ ATPase activity in response to salinity consistent with that of other osmoregulating euryhaline invertebrates. To examine salinity tolerance, growth rates between 5 and 60 ppt were determined. The activity of Na⁺/K⁺ ATPase was examined, over the same range of salinities, by measuring ATPase activity in rotifer homogenates in the presence and absence of a Na⁺/K⁺ ATPase inhibitor. Maximum specific growth rate (0.95 day^–1) occurred at 16 ppt, highest mean amictic eggs per female (1.41) occurred at 20 ppt, and both parameters decreased rapidly as salinity increased. Egg development time was constant with salinity at 0.92 days. The activity of Na⁺/K⁺ ATPase per milligram protein increased from 3.9 µmol h^–1 at 5 ppt to 6.8 µmol h^–1 at 50 ppt and accounted for 15 and 30% of total ATPase activity, respectively. We suggest that these observations are consistent with increasing stress at high salinities and the occurrence of a hypo-osmoregulatory response. Given the high ATP consumption of Na⁺/K⁺ ATPase at high salinities, it is possible that a proportion of the corresponding decreases in growth rate and egg production are a direct cost of regulation.Communicated by J.P. Thorpe, Port Erin     

Target-site sensitivity in a specialized herbivore towards major toxic compounds of its host plant: the Na+K+-ATPase of the oleander hawk moth (Daphnis nerii) is highly susceptible to cardenolides

The caterpillars of the oleander hawk moth, Daphnis nerii (Linnaeus, 1758) (Lepidoptera: Sphingidae) feed primarily on oleander (Nerium oleander). This plant is rich in cardenolides, which specifically inhibit the Na⁺K⁺-ATPase. Since some insects feeding on cardenolide plants possess cardenolide-resistant Na⁺K⁺-ATPases, we tested whether D. nerii also possesses this strategy for circumventing cardenolide toxicity. To do so, we established a physiological assay, which allowed direct measurement of Na⁺K⁺-ATPase cardenolide sensitivity. Using Schistocerca gregaria, as a cardenolide-sensitive reference species, we showed that D. nerii Na⁺K⁺-ATPase was extremely sensitive to the cardenolide ouabain. Surprisingly, its sensitivity is even higher than that of the cardenolide-sensitive generalist, S. gregaria. The presence or absence of cardenolides in the diet of D. nerii did not influence the enzyme’s cardenolide sensitivity, indicating that target-site insensitivity is not inducible in this species. However, despite the sensitivity of their Na⁺K⁺-ATPase, caterpillars of D. nerii quickly recovered from an injection of an excessive amount of ouabain into their haemocoel. We conclude that D. nerii possesses adaptations, which enable it to feed on a cardenolide-rich diet other than that previously described in cardenolide specialized insects, and discuss other potential resistance mechanisms.     

Branchial Na+−K+-ATPase activity during osmotic adjustments in two freshwater euryhaline teleosts,tilapia (Sarotherodon mossambicus) and orange chromid (Etroplus maculatus)

Effects of osmotic stress on the branchial Na⁺–K⁺-ATPase activity in two freshwater euryhaline teleosts (the tilapia Sarotherodon mossambicus Peters and the orange chromid Etroplus maculatus Bleeker) were studied. Direct transfer from fresh water to salt water with a salinity of 35 S caused extensive mortality in tilapia. Of the remaining four saltwater concentrations (4.375, 8.75, 17.5, and 26.25 S), the higher two increased the enzyme activity significantly in the first week without affecting it further during the remaining seven weeks of acclimation. The lower two concentrations failed to elicit any significant change. In the case of orange chromid, all three salt-water concentrations (4.375, 8.75, and 17.5 S) which the fish survived produced a more significant and extensive change in the enzyme activity, which continued to rise during the entire acclimation period. Employing a procedure of gradual transfer, tilapia and orange chromid were successfully adapted to saltwater concentrations of up to 61.25 and 35 S, respectively. In this experiment, almost each successive increase in salinity elevated the enzyme activity further, but the total change in both species was considerably less extensive than might be expected from the direct transfer study. When the freshwater fish were transferred to identical concentrations of pure NaCl or whole ocean salt, the former produced a relatively greater change in the enzyme activity in tilapia. No such difference was seen in orange chromid. During reacclimation of the saltwater-adapted fish to fresh water, the enzyme activity began to decrease immediately in both species, but remained above the freshwater control levels at the end of eight weeks.     

Ecotoxicity of silver nanoparticles on earthworm Eisenia fetida: responses of the antioxidant system,acid phosphatase and ATPase

Ecotoxicity of nanoparticles has received growing attention in recent years. This study investigated the influence of silver nanoparticles (Ag-NP) on earthworm Eisenia fetida. The experiment was performed with five test groups: control (without Ag-NP), 10?nm Ag-NP groups (20, 100 or 500?mg?kg^?1) and positive control (787?mg?kg^?1 AgNO₃). After 14-day acute exposure, activities of various enzymes, including glutathione S-transferase (GST), glutathione reductase (GR), acid phosphatase (AP), and Na⁺, K⁺-ATPase were determined. Effects of Ag-NP with different sizes (10 and 80?nm) were also tested. Data showed that the activity of GR was significantly lower at 500?mg?kg^?1. The activities of AP and Na⁺, K⁺-ATPase were inhibited following the increase of Ag-NP concentration. When treated with Ag-NP with different sizes, activities of AP and Na⁺, K⁺-ATPase of the 10?nm group were significantly lower than the control group, but those of the 80?nm group were similar to the control group. Data indicate that Ag-NP may be harmful to the earthworm E. fetida at 500?mg?kg^?1, and the toxicity of Ag-NP with 10?nm size is greater than 80?nm. In addition, AP and Na⁺, K⁺-ATPase are sensitive biomakers to the effects of Ag-NP.     

Aspects of osmotic and lonic regulation in two Baltic teleosts: Effects of salinity on blood and urine composition

Hydromineral regulation was studied by examining the response to different environmental salinities in two Baltic brackish-water (BW) teleosts—a species of marine ancestry, Myoxocephalus scorpius (L.), and a glacial relict, M. quadricornis (L.). M. scorpius tolerated fresh water (FW) and M. quadricornis sea water (SW) for only about 24 h, but the survival time of M. scorpius in SW and M. quadricornis in FW was one to several weeks. M. scorpius seems able to balance plasma ionic concentrations in salinities down to about 2 to 3. Death of M. scorpius in FW was associated with partial haemolysis, increased volume of red blood cells (RBC), increased plasma K⁺ concentration, and decreased concentrations of Na⁺, Cl^- and Mg²⁺ in plasma and, to a lesser extent, in urine. Death of M. quadricornis in SW was associated with increased plasma osmolality, and Na⁺, Cl^- and Mg²⁺ concentrations, but the renal excretion of ions approached that generally found in marine teleosts. In most cases, RBC volume followed the changes in plasma osmolality or Na⁺ and Cl^- concentrations. Both species showed an ability to increase tubular Mg²⁺ secretion much over that needed in BW, and increased secretion was associated with high urine Cl^- concentration. M. quadricornis, but not M. scorpius, reabsorbed Na⁺ effectively in SW also. Differences between Oceanic and Baltic specimens of M. scorpius are discussed.     

Interaction of alcohol and kola nut on brain sodium pump activity in Wistar rats

The effect of interaction of alcohol and kola nut on sodium pump activity was studied in Wistar rats. Thirty Wistar rats were divided into six groups of five rats per group. Control received a placebo (4 mL of distilled water). Groups 2–6 were treated for a period of 21 days with 10% (v/v) alcohol, 50 mg caffeine/kg, 50 mg kola nut/kg, or a combination of 10% (v/v) alcohol + 50 mg kola nut/kg, or 10% (v/v) alcohol + 50 mg caffeine/kg, respectively. One day after the final exposure, brains were harvested and several biochemical parameters examined including activities of total ATPase, ouabain-insensitive ATPase, ouabain sensitive ATPase (Na⁺–K⁺-ATPase) and levels of nonenzymatic breakdown of ATP and inorganic phosphate (P_i) released. Results showed that the essential enzyme of the brain responsible for neuronal function, Na⁺–K⁺-ATPase, was inhibited by alcohol–kola nut co-administration relative to control, resulting in a decreased ATP production, ion transport and action potential, leading to loss of neuronal activities.     

Amphibian myiasis. Blowfly larvae (Lucilia bufonivora,Diptera: Calliphoridae) coping with the poisonous skin secretion of the common toad (Bufo bufo)

The blowfly Lucilia bufonivora shows high host specificity for toads despite the host’s toxic skin secretion, which consists mainly of bufadienolides. These toxins are effective blockers of the Na⁺, K⁺-ATPase, an enzyme that is essential for many physiological processes in animals. Whereas common toad (Bufo bufo) toxins were identified in the larvae of the fly, few toxins were found in the pupae and empty puparia as trace amounts, while adult flies were entirely free of these toxic compounds. Similar results were obtained when larvae of generalist necrophagous blowflies (L. sericata, Calliphora vicina) fed on toad carcasses. Analysis of the Na⁺, K⁺-ATPase gene revealed no amino acid substitution at positions known to mediate resistance to bufadienolides in other systems. Alternative mechanisms of resistance such as efficient excretion of the compounds may enable the flies to use this poisonous food source.     

Gill and intestinal Na+-K+ ATPase activity, and estimated maximal osmoregulatory costs, in three high-energy-demand teleosts: yellowfin tuna (Thunnus albacares), skipjack tuna (Katsuwonus pelamis), and dolphin fish (Coryphaena hippurus)

We hypothesize that the morpho-physiological adaptations that permit tunas to achieve maximum metabolic rates (MMR) that are more than double those of other active fishes should result in high water and ion flux rates across the gills and concomitant high osmoregulatory costs. The high standard metabolic rates (SMR) of tunas and dolphin fish may, therefore, be due to the elevated rates of energy expenditure for osmoregulation (i.e. teleosts capable of achieving exceptionally high MMR necessarily have SMR). Previous investigators have suggested a link between activity patterns and osmoregulatory costs based on Na⁺-K⁺ ATPase activity in the gills of active epipelagic and sluggish deep-sea fishes. Based on these observations, we conclude that high-energy-demand fishes (i.e. tunas and dolphin fish) should have exceptionally elevated gill and intestinal Na⁺-K⁺ ATPase activity reflecting their elevated rates of salt and water transfer. To test this idea and estimate osmoregulatory costs, we measured Na⁺-K⁺ ATPase activity (V _max) in homogenates of frozen samples taken from the gills and intestines of skipjack and yellowfin tunas, and the gills of dolphin fish. As a check of our procedures, we made similar measurements using tissues from hybrid red tilapia (Oreochromis mossambicus ×O. niloticus). Contrary to our supposition, we found no difference in Na⁺-K⁺ ATPase activity per unit mass of gill or intestine in these four species. We estimate the cost of osmoregulation to be at most 9% and 13% of the SMR in skipjack tuna and yellowfin tuna, respectively. Our results, therefore, do not support either of our original suppositions, and the cause(s) underlying the high SMR of tunas and dolphin fish remain unexplained. Received: 7 September 2000 / Accepted: 4 December 2000