Ion transport in crab gills: A new method using isolated half platelets of Eriocheir gills in an ussing-type chamber

• 1.1. A half platelet preparation from Chinese crab (Eriocheir sinensis) gill is described which allows electrophysiological investigations of ion transport by gill epithelial monolayer when mounted in a modified Ussing chamber.
• 2.2. The resistance of these preparations equals half that of complete gill platelets (containing the gill epithelium and cuticle twice) indicating that cell damage during preparation of half platelets is negligible.
• 3.3. The transepithelial resistance (resistance of cuticle subtracted previously) was determined to be about 140 Ω cm² when both sides are bathed with identical salines.
• 4.4. Similarities to the results obtained with perfused complete gills demonstrates the reliability of this preparation.
• 5.5. When identical salines are applied on both sides of the epithelium an outside positive transepithelial potential difference (PD_te) up to 40 mV was measured.
• 6.6. The occurrence of such a high PD_te under symmetric conditions and its sensitivity to CN⁻ suggests the PD_te to be generated by active transport processes.

     

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.

     

Evidence for hyporegulation in the calanoid copepod,Acartia tonsa

• 1.1. The euryaline calanoid copepod, Acartia tonsa, maintains haemolymph Na below that of the external medium in salinities above 34^o_oo (475 mM Na).
• 2.2. The measured transepithelial electrical potential. −9.97 ± 1.0 mV, indicates that Na is regulated out of electrochemical equilibrium.
• 3.3. Water osmotically lost in hyporegulation is replaced by Na-dependent absorption by the gut.
• 4.4. High osmotic water permeability is evidenced by the fact that with an increase in external salinity from 475 mM Na to 580 mM Na the copepod's drinking rate nearly doubles.
• 5.5. Sodium efflux measurements indicate that ionic permeability is much lower than other hyporegulating crustaceans.
• 6.6. The energetic advantage of hyporegulation in this species is considered.

     

Regulation of amiloride-sensitive Na+ absorption in the lizard (gallotia galloti) colon by aldosterone

• 1.1. Acute (AT) or chronic (CT) administration of exogenous aldosterone brought about a considerable increase in transmural potential difference (PD), short-circuit current (Isc) and net sodium flux (J_net^Na).
• 2.2. A good relationship between Isc and J_m-s^Na was observed in CT but not in AT or UC (untreated controls).
• 3.3. Addition of mucosal amiloride reduced Isc in AT and CT colons, but did not cause any change in UC colons. The relationship between Isc and J_m-s^Na, observed in CT was totally suppressed by the diuretic.
• 4.4. J_net^Na was reduced in AT tissues and abolished in CT colons by amiloride.
• 5.5. The present results strongly suggest that aldosterone levels quantitatively and qualitatively modify sodium absorption across colonie mucosa in a dose-dependent fashion and that lower levels of the hormone are required to induce the electrogenic Na absorption than to suppress the electroneutral transport.

     

Bioelectrical potentials across the mantle epithelium of Achatina fulica

• 1.1. The shell side of the mantle of Achatina fulica is several millivolts positive to the blood side in vitro.
• 2.2. The electrical potential does not depend on Na⁺, Ca²⁺, Mg²⁺, K⁺ or HCO₃⁻ but requires the presence of chloride on the shell side.
• 3.3. The potential difference and short-circuit current ranged from 3.0 to 30.0 mV and 15.0 to 75 μA/cm² with averages at 10m V and 50 μA/cm² respectively.
• 4.4. The electrical gradient is reduced by 2,4-dinitrophenol, thiocyanate and furosemide but not by ouabain, CO₂ or acetozolamide.
• 5.5. It is suggested that the nature and mechanism of electrogenesis in Achatina parallels that of the Helix mantle.

     

Characterization of NaK ATPase from the gills of the freshwater prawn Macrobrachium rosenbergii (de Man)

• 1.1. The specific activity of Na-K ATPase was determined from the microsomal preparation of gills dissected from adult Macrobrachium rosenbergii.
• 2.2. Maximal ATPase activity was achieved at a substrate concentration of 0.5 mM ATP.
• 3.3. Optimal enzyme activity was obtained at pH of 7.5.
• 4.4. The Arrhenius plot of Na-K ATPase activity revealed a marked discontinuity at 30°C. “Mg” ATPase activity did not exhibit a marked discontinuity.
• 5.5. The E_a for Na-K ATPase and “Mg” ATPase was 14.6 kCal/mole and 9.31 kCal/mole respectively. Q₁₀ values for Na-K ATPase was 2.34 and for “Mg” ATPase 1.65.
• 6.6. ATPase activity and gill homogenate protein concentration exhibited a linear relationship up to 130 μg protein/ml.
• 7.7. Na-K ATPase activity was inhibited by 10⁻³ M ouabain. It was equally inhibited by the removal of K⁺ from the reaction medium.

     

Intracellular calcium in the cells of the outer mantle epithelium of Anodonta cygnea

• 1.1. The intracellular concentration of ionized calcium was measured with double-barrelled ion-sensitive microelectrodes under short-circuit conditions in the isolated outer mantle epithelium of Anodonta cygnea.
• 2.2. When the outside baths contained 1 mmol/1 Ca²⁺ the average intracellular Ca²⁺ was 5.42 ± 0.64 mmol/1(N = 41) while the equilibrium concentration estimated from the intracellular potential measured in the same cells was 5.51 ± 0.33 mmol/l.
• 3.3. Bilateral removal of calcium from the external baths induced a fast fall in the intracellular concentration of this ion by almost three orders of magnitude. This effect was similar to that obtained by removing calcium from the bath on the basolateral side.
• 4.4. Removal of calcium from the bath in contact with the apical side of the preparation had little effect on intracellular calcium.

     

Some properties of ATPase activity in the intact cells of yeast Saccharomycopsis fibuligera

• 1.1. The properties of ATPase activity were studied with the cells at the early stationary phase of Saccharomycopsis fibuligera.
• 2.2. Optimal pH for the activity was approximately 7.
• 3.3. The activity was stimulated by Mg²⁺.
• 4.4. The activity was inhibited by NaF, DCCD, oligomycin, NaN₃, NaVO₃, or PCMB but not inhibited by ouabain.

     

The effect of myo-inositol on the ability of Ditylenchus dipsaci,D. myceliophagus and Anguina tritici to survive desiccation

• 1.1. The effect of myo-inositol on the ability of three species of nematodes to survive desiccation has been studied.
• 2.2. Survival rates obtained from worms treated with an inositol bathing medium were compared with survival rates of worms treated with distilled or tapwater media.
• 3.3. Highest survival rates were found in those nematodes that were placed in an inositol solution prior to desiccation.
• 4.4. Tapwater facilitated higher revival rates than did distilled water in both D. dipsaci and D. myceliophagous.
• 5.5. No such differences were found for A. tritici.
• 6.6. The results are discussed in relation to the possible mechanisms of protection afforded by the different bathing media.

     

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.

     

The neural control of release of hyperlipaemic hormone from the corpus cardiacum of Locusta migratoria

• 1.1. The neural control of the release of hyperlipaemic hormone has been studied in isolated corpora cardiaca of Locusta migratoria using electrical stimulation of the nervi corporis cardiaci I and II, and assaying the bathing medium for hyperlipaemic activity.
• 2.2. Stimulation of nervus corpus cardiacum II (NCC II) results in the release of hormone which is dependent upon the arrival of compound action potentials within the glandular lobe of the corpus cardiacum.
• 3.3. Sodium deficient Ringer and Ringer containing 10⁻⁶ M tetrodotoxin abolish the compound action potential and with it the release of hormone.
• 4.4. The mechanism of release is dependent upon the presence of extracellular calcium-ions.
• 5.5. Stimulation of nervus corpus cardiacum I (NCC I) alone does not result in the release of hyperlipaemic hormone, but the presence of action potentials in NCC I potentiates the hyperlipaemic effect obtained by NCC II stimulation.

     

Purification and properties of glycollate oxidase from Lemna minor L.

• 1.1. Glycollate oxidase has been purified to apparent homogeneity from Lemna minor L. grown on medium containing 7mM NO⁻₃.
• 2.2. The enzyme is a highly basic protein with a sub-unit molecular weight of 42,000 and a holoprotein molecular weight of 250,000.
• 3.3. The Lemna enzyme is a flavoprotein with a broad specificity for straight chain α-hydroxy acids, the preferred substrate being glycollate.
• 4.4. It is also competitively inhibited by oxalate and phenyllactate.
• 5.5. A comparison is drawn between the physical properties of glycollate oxidase from a number of higher plants and the degree of sub-unit aggregation in the resulting protomers.

     

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.

     

Purification and properties of tetralysine endopeptidase from Escherichia coli AJ005

• 1.1. A new tetralysine endopeptidase from Escherichia coli AJ005 has been purified about 135-fold.
• 2.2. The peptidase seems to be specific to tetralysine among lysine homopolymers.
• 3.3. The optimal pH was about 7.5
• 4.4. The activity was inhibited by KCN but not inhibited by soybean trypsin inhibitor.
• 5.5. The apparent K_m value was 2.5 × 1O⁻³ M for tetralysine.

     

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.

     

A simple and reliable method for the purification of Pseudomonas aeruginosa phospholipase C produced in a high phosphate medium containing choline

• 1.1. Pseudomonas aeruginosa phospholipase C from culture supernatants of bacteria grown in high-P_i basal salt medium with choline, as the sole carbon and nitrogen source, was purified by precipitation with 70% saturation ammonium sulfate in the presence of celite.
• 2.2. The PLC activity was eluted of this mixture by the use of a reverse gradient of 70-0% ammonium sulfate.
• 3.3. The peak containing the PLC activity revealed a single protein after SDS-PAGE.
• 4.4. The method could also be applied to purify PLC produced in a low-P_i complex medium. The resultant preparation was not homogeneous.
• 5.5. The molecular weight for both PLC preparations was about 70 kDa.
• 6.6. Both PLC used phosphatydilcholine and sphingomyelin as substrates, displayed hemolytic activity an exhibited an apparent K_M of 25 mM for p-nitrophenylphosphorylcholine.
• 7.7. They were not inhibited by 1% sodium deoxycholate but were 30% inhibited by 1% Triton X-100.
• 8.8. 2% sodium dodecylsulfate and 1% tetradecyltrimethylammonium bromide inhibited the PLC from the HP_l-BSM plus choline but not the enzyme from the LP_l-CM.

     

Varying effects of calcium on the oxidation of palmitate and α-ketoglutarate in isolated rat liver mitochondria incubated in KCl-based and sucrose-based media

• 1.1. Isolated mitochondria from rat liver were incubated in the presence of [U-¹⁴C]palmitate, ATP, CoA, carnitine, EGTA (ethylene glycol bis (β-aminoethyl ether) N,N′-tetraacetic acid) and varying amounts of calcium.
• 2.2. When a KCl-based incubation medium was used, the oxidation of palmitate was inhibited when the concentration of free calcium was increased from about 0.1–10μM.
• 3.3. When a sucrose-based incubation medium was used, the basal rate of palmitate oxidation was about half of that observed with the KCl-medium and calcium had a stimulatory effect.
• 4.4. With the KCl-medium the rate of oxygen consumption was inhibited by calcium with α-ketoglutarate as well as palmitate as the respiratory substrate.
• 5.5. No inhibitory effect of calcium was observed with succinate or β-hydroxybutyrate.
• 6.6. With the KCl-medium and with α-ketoglutarate as the respiratory substrate, state 3 respiration but not state 4 respiration was inhibited by calcium.
• 7.7. When the sucrose-medium was used, state 3 respiration was first inhibited by calcium, but this inhibition was gradually relieved and the respiratory rate finally became higher than it was before calcium addition.

     

Na+-independent sugar uptake by rat intestinal and renal brush border and basolateral membrane vesicles

• 1.1. Uptake of [¹⁴C]-labelled d-glucose, l-arabinose and d-fructose by intestinal and renal brush border and basolateral membrane vesicles was studied in the absence of Na⁺ .
• 2.2. The Na⁺-independent d-glucose transport system in these membrane vesicles was saturable, sensitive to phloretin, stereospecific and accessible only to d-glucose and d-galactose.
• 3.3. Na⁺-independent l-arabinose transport was not saturable even when its concentration was raised to 300 mM and it was insensitive to phloretin.
• 4.4. Na⁺-independent d-fructose transport demonstrated saturation kinetics with only renal brush border membrane vesicles, but it was not inhibited by either phloretin or phlorizin.
• 5.5. These studies indicated that the Na⁺-independent carrier-mediated d-glucose/d-galactose transport system of intestinal and renal brush border and basolateral membranes is clearly not shared by other monosaccharides.

     

Na+ regulation and (Na++ K+)ATPase activity in the euryhaline fiddler crab Uca minax (Le conte)

• 1.1. Specific activity and kinetic characteristics of the (Na⁺ + K⁺)ATPase have been investigated in the gill epithelium of the hyper-hypoosmoregulator crab Uca minax.
• 2.2. (Na⁺ +K⁺)ATPase activity is shown to be at least three times higher in the posterior gills.
• 3.3. The kinetic study supports the hypothesis of the existence of two different (Na⁺ + K⁺)ATPases: the enzyme activity in the posterior gills could be involved in the transepithelial transport of Na⁺ while the activity of the anterior gills could be responsible for the intracellular regulation of Na⁺ and K⁺.
• 4.4. Significant and specific changes in (Na⁺ +K⁺)ATPase activity occur upon acclimation to media of various salinities.

     

Effects of Na+ and/or K+ on the Mg2+-dependent ATPase activities in shrimp (Macrobrachium amazonicum) gill homogenates

• 1.1. Homogenates of gills from the freshwater shrimp M. amazonicum exhibit the following ATPase activities: (i) a basal, Mg²⁺-dependent ATPase; (ii) an ouabain-sensitive, Na⁺ + K⁺-stimulated ATPase; (iii) an ouabain-insensitive, Na⁺-stimulated ATPase; and (iv) an ouabain-insensitive, K⁺-stimulated ATPase.
• 2.2. K⁺ suppresses the Na⁺-stimulated ATPase activity in a mixed-type kind of inhibition, whereas Na⁺ does not exert any noticeable effect on the K⁺-stimulated ATPase activity.
• 3.3. The Na⁺- and the K⁺-stimulated ATPase activities are totally inhibited by 5 mM ethacrynic acid in the incubation medium.
• 4.4. The Na⁺- and the K⁺-stimulated ATPase activities are not expressions of the activation of a Ca-ATPase.
• 5.5. The possible localization and roles of the described ATPases within the gill epithelium are briefly discussed and evaluated.