Sea-water tolerance in freshwater-resident arctic charr (Salvelinus alpinus)

• 1.1. Freshwater-resident Arctic charr acclimated for 2 months at 8°C, 15% were divided into four experimental groups in July and exposed to 1 and 8°C in 15 and 34% salinity.
• 2.2. Only slight changes in gill Na-K-ATPase activity, blood plasma osmolality and blood plasma concentrations of Cl⁻ and Mg²⁺ were found for the fish exposed to 1 or 8°C in brackish water.
• 3.3. When exposed to sea-water at 8°C, an increase in osmolality and in concentrations of Cl⁻ and Mg²⁺ took place during the first 2–3 days, after which it levelled off.
• 4.4. If exposed to sea-water at 1°C, however, marked increases were found for all parameters measured and all the fish were dead within 5 days of exposure.
• 5.5. These results show that freshwater-resident Arctic charr—if acclimated to brackish water—can survive in sea-water during summer if the environmental temperature is not too low.

     

Sodium transport in red blood cells of lamprey LAmpetra fluviatilis

• 1.1. Unidirectional Na⁺ influx in lamprey red blood cells was determined using ²²Na as a tracer.
• 2.2. Total Na⁺ uptake and amiloride-inhibitable Na⁺ influx increased in a saturable fashion as a function of external Na⁺ concentration (Na_e).
• 3.3. At 141 mM Na_e, the average value of net Na⁺ influx was 13 ± 1.1 and the amiloride-sensitive Na⁺ influx was 5.3±1.1 mmol/l cells per hr (±SE).
• 4.4. The amiloride-sensitive component of Na⁺ influx was significantly activated by 10⁻⁵ M isoproterenol, by 2 × 10⁻⁵ M DNP, and by cell shrinkage.
• 5.5. Furosemide (1 mM) had no effect on the Na⁺ transport in red cells.
• 6.6. The residual amiloride-insensitive component of Na⁺ transport was a linear function of Na_e in the range of 5–141 mM. This transport seems to be accounted for by simple diffusion.

     

Intestinal l-methionine transport in the cultured gilthead bream (Sparus aurata)

• 1.1. The influx and transepithelial movements of l-methionine and its effects on the electrophysiology and Na-Cl-transport in upper and lower intestine of the cultured fish, Spanis aurata, were measured.
• 2.2. The K_m and V_max of l-methionine influx into the tissues were higher in lower intestine than in upper intestine. A prominent diffusion-like transport component was also measured in both segments during influx experiments.
• 3.3. Net transepithelial fluxes of l-methionine (1 mM) were observed in both upper and lower intestine, this transport being Na⁺-dependent.
• 4.4. The two intestinal segments exhibited an electrical potential difference (PD) and a short circuit current (I_sc) serosa negative or near zero. Tissue conductance (G_t) was higher in posterior than in lower intestine.
• 5.5. Addition of l-methionine to the mucosal side of lower or upper intestine did not induce changes in PD in either part.
• 6.6. Isotopic fluxes of Cl⁻ or Na⁺ measurements under short circuit conditions showed that there were no net Cl⁻ or Na⁺ transport in either part.
• 7.7. l-Methionine additions to the mucosa did not induce changes in unidirectional fluxes of Cl⁻ or Na⁺ or in the (I_sc) in either the anterior or posterior intestine.

     

Effects of aluminium and pH on calcium fluxes,and effects of cadmium and manganese on calcium and sodium fluxes in brown trout (Salmo trutta L.)

• 1.1. Simultaneous measurement of calcium fluxes in brown trout, at low external [Ca] (20 μ mol 1⁻¹), provided evidence of active uptake of Ca from the medium.
• 2.2. At pH 4.5, calcium influx was inhibited and efflux was stimulated.
• 3.3. Cd and Mn, but not Al, at concentrations within the ranges found in acid waters experiencing fish population decline, inhibited calcium influx. Efflux was unaffected.
• 4.4. Cd and Mn stimulated sodium influx and efflux.

     

Nacl flux in the flounder (Pseudopleuronectes americanus) intestine: Effects of ph and transport inhibitors

• 1.1. The effects of extracellular pH on Na⁺ and Cl⁻ absorption were studied in vitro in the small intestine of the winter flounder, Pseudopleuronectes americanus.
• 2.2. Reductions in bathing solution pH inhibited J_ms^na (mucosal-to-serosal flux) and J_net^na (net flux) (r = 0.90) and J_net^Cl (r = 0.92) [due to an increase in J_sm^Cl, (serosal-to-mucosal)] and decreased short circuit current (Isc).
• 3.3. Luminal bumetanide (0.1 mM) and amiloride (1 mM) inhibited Na⁺ and Cl⁻ absorption by reducing J_ms.
• 4.4. Luminal barium (5mM) and luminal copper (100 μM) decreased J_ms^Cl and increased J_sm^Cl.
• 5.5. We conclude that reductions in extracellular pH inhibit a luminal membrane NaCl absorptive process (Na⁺-K⁺-2Cl⁻) and stimulate an electrogenic Cl⁻ secretory process.

     

The effects of heavy metals and metabolic inhibitors on calcium uptake by gills and scales of Fundulus heteroclitus in vitro

• 1.1. Cadmium (Cd) and zinc (Zn) were inhibitory to calcium uptake by isolated gills of Fundulus heteroclitus in vitro. The metals appeared to act by displacing Ca²⁺ ions from protein carriers involved in facilitated diffusion.
• 2.2. In saltwater fish, transport of calcium across the serosal membrane of gill chloride cells is partly energy dependent and is likely mediated by Ca²⁺-ATPase. However, much of the calcium transport through the gill epithelium appears to occur by passive processes.
• 3.3. Cd (10⁻⁵M—10⁻³M) and Zn (10⁻⁷M—10⁻³ M) inhibited calcium uptake by isolated scale patches incubated in a physiological saline.
• 4.4. Cyanide, oubain, and quercetin treatment of scale patches produced results similar to those of the Cd and Zn treatments suggesting that metal-induced inhibition of ATPases may be responsible for reduced calcium transport by scale osteoblasts.

     

Cl− fluxes across isolated,perfused gills of the chinese crab Eriocheir sinensis (M.Edw.) acclimated to fresh water

• 1.1. The movements of Cl⁻¹ have been studied in the so-called anterior and posterior gills of E. sinensis using radioactive ³⁶Cl⁻¹.
• 2.2. The anterior gills hardly show any significant movements of Cl⁻¹. They thus have a very low (if any) permeability to that ion. On the contrary, the posterior gills show both passive fluxes and an active inward movement of Cl⁻¹.
• 3.3. The Cl⁻¹ influx in the posterior gills is largely sensitive to the amount of K⁺ in the perfusion saline.

     

Evidence for extracellular deamination of adenosine in the rat heart

• 1.1. In rat heart perfused with adenosine (10⁻⁶M), dilazep (10⁻⁴M) inhibited incorporation of adenosine into nucleotides (an index of nucleoside transport and phosphorylation) to a greater extent (70%) than metabolism to inosine and uric acid (40%) and actually increased the recovery of inosine to 30% of the adenosine infused.
• 2.2. Extrapolating for complete inhibition of transport suggested that 60% of adenosine metabolism was intracellular and 40% extracellular.
• 3.3. Static incubations of atria also gave an estimate for extracellular metabolism of 40%.
• 4.4. Adenosine deaminase was localised by immunocytochemistry to the extracellular surface of endothelial cells of small coronary arteries.
• 5.5. Extracellular deamination may explain the lack of effect of nucleoside transport inhibitors on responses to adenosine in rat heart.

     

Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. V—Rabbit (Oryctolagus Cuniculus)

• 1.1. The diffusional water permeability (P_d) of rabbit red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulfonate (PCMBS).
• 2.2. The values of P_d were around 6.3 × 10⁻³ cm/sec at 15°C, 7.0 × 10⁻³cm/sec at 20°C, 8.0 × 10⁻³ cm/sec at 25°C, 9.1 × 10⁻³ cm/sec at 30°C and10.7 × 10⁻³ cm/sec at 37°C.
• 3.3. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 15 min at 37°C with 0.5 mM PCMBS.
• 4.4. The values of maximal inhibition were around 71–74% at all temperatures.
• 5.5. The basal permeability to water was estimated as 1.6 × 10⁻³cm/sec at 15°C, 2.0 × 10⁻³cm/sec at 20°C, 2.4 × 10⁻³cm/sec at 25°C, 2.6 × 10⁻³cm/sec at 30°C, and 3.1× 10^{−3 cm/secat 37°C}.
• 6.6. The activation energy of water diffusion was around 18 kJ/mol and increased to 27 kcal/mol after incubation with PCMBS in conditions of maximal inhibition of water diffusion.
• 7.7. The membrane polypeptide electrophoretic pattern of rabbit RBCs has been compared with its human counterpart.
• 8.8. The rabbit membrane contained a higher amount of spectrin (bands 1 and 2), while the band 6 (glyceraldehyde-3-phosphate dehydrogenase) was markedly less intense.
• 9.9. Considerable differences in the electrophoretic patterns of the two sources of RBC membranes appeared in the bands migrating in the band 4.5 region and in front of band 7, where some polypeptides were apparent in higher amounts in the rabbit RBC membrane.

     

Cationic and anionic channels of apical receptive membrane in a taste cell contribute to generation of salt-induced receptor potential

• 1.1. After ionic composition of superficial fluid (ISF) and interstitial fluid (ISF) of the frog Rana catesbeiana) tongue had mostly been changed with a low Na⁺ saline solution, the relations between membrane potentials and receptor potentials in a frog taste cell evoked by various concentrations of NaCl and various types of salts were analyzed to examine permeability of the taste receptive membrane to cations and anions.
• 2.2. The mean reversal potentials for depolarizing potentials of a taste cell in response to 0.05 M, 0.2 M and 0.5 M Nad were -40.0, 6.4 and 28.8 mV, respectively.
• 3.3. When adding an anion channel blocker, SITS, to a NaCl solution the reversal potential for receptor potential with NaCl plus SITS became about twice as large than with NaCl alone.
• 4.4. Reversal potentials for 0.2 M NaCl, LiCl, KCl and NaSCN were 6.4, 25.4, −1.0 and −7.8 mV, respectively, indicating that permeability of the apical taste receptive membrane to cations of Cl⁻ salts is arranged in the order of Li⁺ > Na⁺ > K⁺ and that the permeability to anions of Na⁺ salts is arranged as SCN⁻ > Cl⁻
• 5.5. It is concluded that in the case of NaCl stimulation, Na⁺ and Cl⁻ of NaCl stimulus permeate NaCl-gated cationic and anionic channels at the apical taste receptive membrane in generating receptor potentials.

     

Driving forces for the uphill transport of amino acids into epidermal brush border membrane vesicles of the sea anemone,Anemonia sulcata (Cnidaria,anthozoa)

• 1.1. The transport of amino acids into membrane vesicles prepared from epidermal tentacle tissue of the sea anemone, Anemonia sulcata, depends on an electrochemical potential difference caused, e.g. by sodium chloride gradients.
• 2.2. Potassium or choline chloride gradients energized the transport less effectively than sodium chloride gradients. Both Na⁺-ions and Cl⁻-ions were required for the amino acid transport.
• 3.3. The uphill transport of amino acids along the downhill movement of driver ions (sodium chloride gradient conditions) was characterized by an overshoot; under sodium chloride equilibrium conditions, however, an accumulation of amino acids within the vesicles could not be measured.
• 4.4. Potassium diffusion potentials in combination with valinomycin indicated that hyperpolarization (vesicle inside negative) and hypopolarization (vesicle inside positive) enhanced or depressed the accumulation of amino acids within the vesicles.
• 5.5. Being at the phylogenetic base of the Eumetazoa, cnidarians show characteristics for the transmembrane transport of amino acids comparable to those established for vertebrates.

     

Changes in composition and proteolytic enzyme activities of artemia during early development

• 1.1. The proximate composition, total and free amino acids, and proteases of Artemia nauplii were determined during early development.
• 2.2. Moisture increased from 71.0% to 80.8%, crude protein decreased from 13.2% to 8.8%, crude fat and ash varied slightly.
• 3.3. The total amino acids decreased. Free amino acids changed in three patterns.
• 4.4. Trypsin, chymotrypsin, carboxypeptidase A, B and cathepsin B and C increased in activity. The activity of trypsin was lower, while cathepsin B and C were the highest.
• 5.5. The protease activities were maximal at pH 7.5 and 8.0, and at 45°C on casein.
• 6.6. The optimal pH for carboxypeptidase A was 4.0, for carboxypeptidase B was 4.5, for trypsin and chymotrypsin were 7.0–7.5. The protease(s) active at pH 9.0–9.5 were to be determined.

     

Amino acid sequences around exofacial proteolytic cleavage sites of band 3 from bovine and porcine erythrocytes

• 1.1. Amino acid sequences of bovine and porcine band 3, an erythrocyte anion transporter, were determined.
• 2.2. The sequence of bovine band 3 was positioned to residues 519–599 (the numbering is based on human band 3), in which probably 6 residues were unidentified.
• 3.3. Binding site of DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonate), a potent anion transport inhibitor, was identified as Lys-539 in the bovine case.
• 4.4. A loop (residues 551–567), which provides exofacial proteolytic cleavage sites, contains only 53% homology between human and bovine, whereas the residues flanking it on either side are >84% homologous.
• 5.5. Furthermore, the loop of porcine band 3 was indicated to consist of a 6 or 7-residues short peptide as compared with those of other species.

     

Measurement of blood flow to osseous tissue in dogs using the radiolabelled microsphere method

• 1.1. The purpose of this study was to examine bone blood flow in various intra- and extra-oral sites.
• 2.2. The radiolabelled microsphere method was utilized to assess osseous blood flow in the following regions of 10 dogs: rib, long bone, and anterior and posterior regions of the maxilla and mandible.
• 3.3. Samples of cancellous and cortical bone were also obtained from each of these regions with the exception of the maxilla and the anterior mandible.
• 4.4. Mean blood flow ranged from 3.71 ±0.81 (SE) ml min.⁻¹ 100 g⁻¹ in the mandibular posterior cortical bone to 22.7±4.66ml min⁻¹ 100 g⁻¹ in the cancellous rib samples.
• 5.5. Blood flow to the cancellous tissue of the rib was significantly greater (P < 0.05 ) than the other tissues with the exception of maxillary posterior bone and cortical rib.
• 6.6. Results from this study indicate that blood flow to the maxillary posterior bone is relatively high, but blood flow in other intraoral osseous sites is significantly less than that of cancellous rib bone.

     

Glucose transport across the intestinal wall of the freshwater snail Biomphalaria glabrata

• 1.1. Isolated midguts of the freshwater snail Biomphalaria glabrata were mounted in an incubation chamber in saline containing 2 mM glucose and perfused with the same solution. External and internal media were continuously gassed with carbogen gas (95% O₂, 5% CO₂). In order to measure the flux rates of glucose [¹⁴C]glucose was applied in the perfusion medium or in the incubation medium. Net fluxes of glucose were calculated as the differences between unidirectional in- and effluxes.
• 2.2. A directed net flux from the mucosal to the serosal side of the intestine was demonstrated (mucosal to serosal = 50 ± 10 nmol cm⁻²hr⁻¹(N = 6) serosal to mucosal 7 ± 1 nmol cm⁻²hr⁻¹ (N = 6), net flux = 43 nmol cm⁻²hr⁻¹).r
• 3.3. The active transport of glucose was reduced by the presence of metabolic inhibitors, cyanide (1 mM) and dinitrophenol (1 mM) on the mucosal as well as on the serosal side. Ouabain (1 mM) inhibited the transport rate only when it was added on the serosal side. Amiloride (1 mM) had no effect on the transport rate whether added on the mucosal or on the serosal side.
• 4.4. Inhibition of glucose transport by oubain, a specific inhibitor of Na⁺/K⁺-ATPase, suggests that glucose transport is secondary active and coupled to Na⁺-transport.

     

Blood changes in japanese eels,Anguilla japonica,experimentally infected with typical or atypicalAeromonas salmonicida

• 1.1. Changes in the blood and in the rate of oxygen consumption of Japanese eels injected intramuscularly in the head with a lethal dose of typical or atypical Aeromonas salmonicida at 20°C were investigated.
• 2.2. Eels infected with the bacteria became moribund within 4 to 6 days, and then died within 1 day.
• 3.3. The O₂ consumption rate and blood parameters changed markedly with infections. The responses of hosts to infection by the two kinds of bacteria differed with regard to the following four points: blood pH, plasma Cl⁻, lactic acid, and the numbers of granulocytes and lymphocytes.
• 4.4. The responses of eels infected with atypical A. salmonicida were larger and more rapid than those of eels infected with typical A. salmonicida.

     

Serum ions concentration and atpase activity in gills,kidney and oesophagus of european sea bass (Dicentrarchus labrax,pisces, perciformes) during acclimation trials to fresh water

• 1.1. Kidney, oesophagus and gill Na⁺-K⁺ ATPase activity and serum Na⁺, K⁺ and Cl⁻ concentrations are evaluated in European sea bass during experimental acclimation to fresh water.
• 2.2. Kidney and oesophagus ATPase increase in low salinity and reach a maximum in fresh water.
• 3.3. Gill ATPase decreases during the acclimation trials and rises again to normal values after a 3-week stay in fresh water.
• 4.4. Na⁺ and K⁺ serum concentrations decrease during the trials and increase back after a 3-week stay in fresh water.
• 5.5. The correlations between enzymatic activities, serum ion concentrations, morphological changes and environmental salinity are discussed.

     

The isolation and partial characterization of α2-macroglobulin from the serum of the ostrich (Struthio camelus)

• 1.1. α₂-Macroglobulin (α₂M) activity is present in the serum of the ostrich, Struthio camelus. The chromogenic synthetic peptide substrates BAPNA and ATNA were hydrolysed by trypsin and chymotrypsin, respectively, in the presence of ostrich serum and the α₂M in ostrich serum protected trypsin from being inhibited by soybean trypsin inhibitor. Ostrich α₂M proved to be a potent inhibitor of bovine pancreatic trypsin and chymotrypsin.
• 2.2. α₂M was purified to apparent homogeneity by PEG precipitation, DEAE-Toyopearl 650M, Bio-Gel A-5m and Zn²⁺-affinity chromatography.
• 3.3. Ostrich α₂M migrated as a single band (M_r 779,000) during non-denaturing gradient gel electrophoresis and showed increased mobility after reaction with trypsin. Denaturation dissociated ostrich α₂ M into half-molecules. Denaturation with reduction further dissociated the protein into quarter-subunits.
• 4.4. Isoelectric focusing revealed a pI of 5.3.
• 5.5. The amino acid composition of ostrich α₂M is typical of an α₂M, comparing favourably with those of other animal species. The carbohydrate composition of the purified protein, in percentage dry weight of the molecule, was galactose: mannose (1:1), 4.55; N-acetylglucosamine, 2.35; N-acetylneuraminic acid, 0.58; and fucose, 0.77.
• 6.6. α₂M was assessed immunologically by Ouchterlony double-diffusion and Western blot analysis with polyvalent antisera directed against ostrich α₂M.
• 7.7. Ostrich α₂M seems to show many physical, chemical and kinetic properties similar to those of other known α₂Ms, but is expected to differ from other αMs when considering the primary structure of the bait region, the area differing among α Ms from different species and determining its specificity.

     

Effect of repetitive cortisol and thyroxine injections on chloride cell number and Na+/K+-ATPase activity in gills of freshwater acclimated rainbow trout,Salmo gairdneri

• 1.1. Freshwater nonanadromous rainbow trout, Salmo gairdneri, were injected three times a week with either saline, 10μg cortisol/g, 1.0μg thyroxine/g or 10μg cortisol/g + 1.0μg thyroxine/g during a period of 28 days (12 injections). A separate group was derived as a subgroup from the thyroxine group on day 14 and received Cortisol + thyroxine from day 14 until day 28 (six injections).
• 2.2. Gill chloride cell number and Na⁺/K⁺-ATPase activity increased by cortisol treatment, the changes being significant on days 7 and 14, respectively.
• 3.3. Thyroxine treatment did not affect gill Na⁺/K⁺-ATPase activity or chloride cell number directly. Neither did it modify the stimulatory effect of cortisol on these parameters.
• 4.4. Muscle water decreased in cortisol-treated fish and increased in thyroxine-treated fish, while no changes were observed in the combined hormone groups.
• 5.5. No changes were observed in plasma chloride in any group during the experiment.
• 6.6. The results demonstrate a putative role of cortisol in stimulating hypo-osmoregulatory mechanisms and suggest that thyroxine is without a direct or a supportive effect for cortisol action.

     

Absorption and blood clearance of labelled carotenoids ([14C]astaxanthin, [3H]Canthaxanthin and [3H]zeaxanthin) in mature female rainbow trout (Oncorhynchus mykiss)

• 1.1. Using one force-fed meal, eight mature female rainbow trout received [¹⁴C]astaxanthin ([¹⁴C]Ax) with [³H]canthaxanthin ([³H]Cx; N = 3) or with [³H]zeaxanthin ([³H]Zx; N = 5).
• 2.2. Approximately 200 μl of blood were collected via caudal puncture every 24 hr for 4 days. After 96 hr, the fish were killed and pyloric caeca (P.C.) from the duodenal intestine (D.I.) section, ileal intestine (I.I.), and posterior intestine (P.I.) were dissected out.
• 3.3. In the blood, Ax levels were higher than Cx followed by Zx levels.
• 4.4. This corresponds to their respective absorption by the trout as was confirmed by their relative concentrations in P.C., I.I. and P.I.
• 5.5. However, blood clearance was similar for all three compounds. [¹⁴C]Phoenicoxanthin ([¹⁴C]Px) was detected as a reduced metabolite of [¹⁴C]Ax in all gut sections.