Two sodium pumps in the hepatopancreas of the intertidal euryhaline crab Neohelice granulata: biochemical characteristics and differential modulation after feeding

2018 ◽  
Vol 96 (6) ◽  
pp. 576-584 ◽  
Author(s):  
E. Méndez ◽  
C. Caruso Neves ◽  
A.A. López Mañanes
Keyword(s):  
Atpase Activity ◽  
Environmental Conditions ◽  
Sodium Pump ◽  
Differential Modulation ◽  
Model Species ◽  
Biochemical Characteristics ◽  
Neohelice Granulata ◽  
Sodium Pumps ◽  
Total Inhibition ◽  
Occurrence Characteristics

No study has been done on the existence, biochemical characteristics, and modulation of K+-independent ouabain-insensitive Na+ ATPase activity (the second sodium pump) in the digestive tract of intertidal euryhaline crabs and moreover on the coexistence and modulation under distinct physiological and (or) environmental conditions of different sodium pumps. We determined the occurrence, characteristics, and responses at different times (0, 1, 24, 48, and 120 h) after feeding upon distinct salinities of Na+ ATPase activity and Na+/K+ ATPase in the hepatopancreas of Neohelice granulata (Dana, 1851), which is a model species. The stimulation by Na+ under total inhibition of Na+/K+ ATPase activity revealed the occurrence of Na+ ATPase activity that was totally inhibited by 2 mmol·L–1 furosemide, exhibits Michaelis–Menten kinetics for ATP (apparent Km = 0.52 ± 0.16 mmol·L–1), and highest activity at around pH 7.4. In crabs acclimated to 35 psu (osmoconforming conditions), Na+ ATPase activity was highly increased (about 15-fold) (532 ± 58 nmol Pi·mg protein−1·min−1) in the hepatopancreas 48 h after feeding. In 10 psu (hyper-regulating conditions), Na+ ATPase activity decreased in the hepatopancreas 24 h after feeding (7 ± 9 nmol Pi·mg protein−1·min−1) and recovered initial values after 48 h (24 ± 35 nmol Pi·mg protein−1·min−1). Unlike Na+ ATPase, Na+/K+ ATPase activity did not change after feeding at any salinity, suggesting the specific modulation of the second sodium pump and its role in postprandial adjustments in the hepatopancreas.

Ouabain binding, Na+-K+-ATPase activity, and 86Rb uptake of canine arteries

1983 ◽  
Vol 245 (4) ◽  
pp. H604-H609
Author(s):  
R. D. Bukoski ◽  
C. L. Seidel ◽  
J. C. Allen
Keyword(s):  
Atpase Activity ◽  
Renal Artery ◽  
Femoral Artery ◽  
Sodium Pump ◽  
Dissociation Constants ◽  
Renal Arteries ◽  
Ouabain Binding ◽  
Pump Function ◽  
Sodium Pumps ◽  
Rate Limiting

In a previous report [Am. J. Physiol. 245 (Heart Circ. Physiol. 14): H598-H603, 1983] we measured potassium-induced relaxation of canine femoral and renal arteries as an index of sodium pump function and concluded that it may not be an accurate measure. In this report, the sodium pumps of femoral and renal arteries were examined using three separate approaches to compare sodium pump function more directly. The number of pump sites in subcellular fractions was measured using [3H]ouabain binding. No differences were observed (femoral artery 14.5 +/- 4.8 pmol/g wet wt; renal artery 17.4 +/- 6.5). Similarly, there was no difference in ouabain-inhibitable Na+-K+-ATPase activity in NaI-treated microsomal fractions of these arteries (femoral artery 3.10 +/- 0.65 mumol Pi X mg-1 X h-1; renal artery 3.42 +/- 0.23). Finally, sodium pump function was measured using the ouabain-inhibitable 86Rb uptake method. The rate of ouabain-inhibitable 86Rb uptake was found to be identical for the two arteries, as were the dissociation constants of the two arteries for rubidium. However, in the presence of 3.5 microM norepinephrine, the rate of 86Rb uptake in the renal artery was greater than that of the femoral artery. We conclude that, under resting conditions, the sodium pumps of femoral and renal arteries are identical, whereas in the presence of norepinephrine, the renal artery sodium pump is not rate limiting with respect to contractility. These data have significant implications with regard to the use of potassium-induced relaxation as an index of sodium pump activity.

Differential modulation after feeding in different salinities and response to abscisic acid (ABA) and extracellular Ca2+ of aminopeptidase N (APN) activity in the hepatopancreas of the intertidal euryhaline crab Neohelice granulata

2020 ◽  
Vol 98 (4) ◽  
pp. 262-268
Author(s):  
M.S. Michiels ◽  
G.R. Daleo ◽  
A.A. López Mañanes
Keyword(s):  
Enzyme Activity ◽  
Abscisic Acid ◽  
Digestive Tract ◽  
Digestive Enzyme ◽  
Aminopeptidase N ◽  
Differential Modulation ◽  
Model Species ◽  
Main Site ◽  
Neohelice Granulata

Modulation of aminopeptidase N (APN) activity in the digestive tract by various factors would be important to adjust digestive and absorptive processes under different physiological and (or) environmental conditions. We studied the postprandial responses at different salinities and the effect of abscisic acid (ABA) and extracellular Ca2+ on APN activity in the hepatopancreas (the main site for nutrient digestion and absorption) of the model species Neohelice granulata (Dana, 1851). Enzyme activity was determined at different times (0, 24, 48, and 72 h) after feeding in crabs acclimated either to 35 psu (osmoconformation) or 10 psu (hyper-regulation). APN activity increased around 50% at 24 h after feeding at 35 psu, whereas no changes occurred at 10 psu. Enzyme activity was also assayed in the presence of ABA (1 × 10–4 mol·L–1) or extracellular Ca2+ (1 × 10–4 mol·L–1), showing increments of 60% and 56%, respectively. The results suggest a role of APN in postprandial adjustments and its modulation by different chemical messengers by direct effect on the hepatopancreas. Moreover, to our knowledge, this work is the first to show the effect of ABA on a digestive enzyme in the digestive tract of an animal.

Metabolic importance of Na+/K+-ATPase activity during sea urchin development.

1997 ◽  
Vol 200 (22) ◽  
pp. 2881-2892 ◽  
Author(s):  
P Leong ◽  
D Manahan
Keyword(s):  
Enzyme Activity ◽  
Atpase Activity ◽  
Sea Urchin ◽  
Sodium Pump ◽  
Sea Water ◽  
Strongylocentrotus Purpuratus ◽  
Lytechinus Pictus ◽  
Stimulation Of

Early stages of animal development have high mass-specific rates of metabolism. The biochemical processes that establish metabolic rate and how these processes change during development are not understood. In this study, changes in Na+/K+-ATPase activity (the sodium pump) and rate of oxygen consumption were measured during embryonic and early larval development for two species of sea urchin, Strongylocentrotus purpuratus and Lytechinus pictus. Total (in vitro) Na+/K+-ATPase activity increased during development and could potentially account for up to 77 % of larval oxygen consumption in Strongylocentrotus purpuratus (pluteus stage) and 80 % in Lytechinus pictus (prism stage). The critical issue was addressed of what percentage of total enzyme activity is physiologically active in living embryos and larvae and thus what percentage of metabolism is established by the activity of the sodium pump during development. Early developmental stages of sea urchins are ideal for understanding the in vivo metabolic importance of Na+/K+-ATPase because of their small size and high permeability to radioactive tracers (86Rb+) added to sea water. A comparison of total and in vivo Na+/K+-ATPase activities revealed that approximately half of the total activity was utilized in vivo. The remainder represented a functionally active reserve that was subject to regulation, as verified by stimulation of in vivo Na+/K+-ATPase activity in the presence of the ionophore monensin. In the presence of monensin, in vivo Na+/K+-ATPase activities in embryos of S. purpuratus increased to 94 % of the maximum enzyme activity measured in vitro. Stimulation of in vivo Na+/K+-ATPase activity was also observed in the presence of dissolved alanine, presumably due to the requirement to remove the additional intracellular Na+ that was cotransported with alanine from sea water. The metabolic cost of maintaining the ionic balance was found to be high, with this process alone accounting for 40 % of the metabolic rate of sea urchin larvae (based on the measured fraction of total Na+/K+-ATPase that is physiologically active in larvae of S. purpuratus). Ontogenetic changes in pump activity and environmentally induced regulation of reserve Na+/K+-ATPase activity are important factors that determine a major proportion of the metabolic costs of sea urchin development.

Dopamine causes inhibition of Na+-K+-ATPase activity in rat proximal convoluted tubule segments

1987 ◽  
Vol 252 (1) ◽  
pp. F39-F45 ◽  
Author(s):  
A. Aperia ◽  
A. Bertorello ◽  
I. Seri
Keyword(s):  
Atpase Activity ◽  
Sodium Pump ◽  
Maximal Activity ◽  
Proximal Convoluted Tubule ◽  
Decarboxylase Activity ◽  
Dependent Manner ◽  
Site Of Action ◽  
Apparent Reduction ◽  
Ly 171555 ◽  
Dose Dependent

We studied the effect of dopamine (DA) on Na+-K+-ATPase activity in proximal convoluted tubule (PCT) segments dissected from perfused rat kidneys. DA inhibited Na+-K+-ATPase activity in a dose-dependent manner. Inhibition was significant with 10(-7) M DA and maximal with 10(-4) M DA. The inhibition was reversible. Enzyme inhibition occurred in the presence of DA and a DA antagonist, metoclopramide, but not when 10(5) M of the DA1 and DA2 agonists fenoldopam mesylate and LY 171555 were added in the absence of DA. In PCT segments incubated with the DA precursor dopa, Na+-K+-ATPase activity was also inhibited. However, dopa did not inhibit the sodium pump if dopa decarboxylase activity was blocked with benserazide. These findings suggest an intracellular site of action of DA. In tubules incubated in different K concentrations, 10(-5) DA decreased the maximal activity (Vmax) and increased the Km. DA 10(-5) M caused an almost immediate swelling of PCT segments. Swelling did not occur in the presence of both DA and 10(-5) M amiloride. The DA-induced tubular swelling was probably due to inhibition of Na+-K+-ATPase-mediated Na+-transport. We conclude that in rat PCT segments, DA causes a rapid and reversible inhibition of apparent Na+-K+-ATPase activity and an apparent reduction in the affinity for K. The site of action appears to be intracellular.

Molecular activity of sodium pumps in endotherms and ectotherms

1996 ◽  
Vol 271 (5) ◽  
pp. R1287-R1294 ◽  
Author(s):  
P. L. Else ◽  
D. J. Windmill ◽  
V. Markus
Keyword(s):  
Skeletal Muscle ◽  
Enzymatic Activity ◽  
Direct Measurement ◽  
Sodium Pump ◽  
Turnover Rates ◽  
Ouabain Binding ◽  
General Increase ◽  
Sodium Pumps ◽  
Invertebrate Species ◽  
Pump Activity

Previous research has shown ectotherms to have markedly lower sodium pump metabolism than endotherms. Direct measurement of enzymatic activity of the sodium pump (Na(+)-K(+)-adenosinetriphosphatase) confirmed this difference. To determine the source of this difference, sodium pump density was measured with the use of [3H]ouabain binding. Ectotherms and endotherms were found to share similar sodium pump numbers. Approximate densities (in pmol/g) were 250 for skeletal muscle, 500 for liver, 900 for heart, and 8,000 for kidney and brain. Therefore, differences in sodium pump activity between endotherms and ectotherms were due to differences in turnover rates or molecular activities of sodium pumps. Molecular activities of sodium pumps (in ATP/min) of tissues from endotherms were between 6,000 and 12,000 and, for ectotherms, between 1,500 and 2,500. Exceptions were found that included the heart of Bufo marinus. In a single invertebrate species studied, Charax destructor, the sodium pumps of the heart had a low molecular activity characteristic of ectothermic tissues. These results suggest that during the evolution of endothermy there was a general increase in the molecular activity of the sodium pump.

Feeding and osmoregulation in the euryhaline crab Neohelice granulata: digestive parameter responses

2019 ◽  
Vol 69 (4) ◽  
pp. 433-449 ◽  
Author(s):  
Eugenia Méndez ◽  
Alejandra A. López Mañanes ◽  
Silvina A. Pinoni
Keyword(s):  
Proteolytic Activity ◽  
Coastal Lagoon ◽  
Environmental Conditions ◽  
Digestive Enzymes ◽  
Buenos Aires ◽  
Amylase Activity ◽  
Metabolic Flexibility ◽  
Buenos Aires Province ◽  
Neohelice Granulata

Abstract When a species often experiences variation in its environmental conditions, metabolic flexibility is required. We studied the duration of the digestive cycle and the activity of key digestive enzymes (proteolytic, amylase, lipase) at short and long times after feeding in the hepatopancreas of the osmoregulator crab Neohelice granulata from the Mar Chiquita coastal lagoon (Buenos Aires Province, Argentina). We compared these responses upon hyper-regulation and under osmoconformation conditions (10 and 35 psu). The results show the ability of this crab to hyper-regulate in different feeding states. No significant differences were observed in the duration pattern of the digestive cycle between hyper-regulation and osmoconformity. However, distinct responses after feeding were observed in the activities of the digestive enzymes studied in relation to the osmoregulatory state. In individuals exposed to 35 psu, proteolytic activity was higher at 8 h and remained constant until 72 h after feeding. At 10 psu, this activity was higher at 48 h after feeding than before feeding. At 35 psu, the amylase activity after feeding was greater than the prefeeding activity. No differences were observed in the activity of lipase at 35 psu, but at 10 psu this activity was lower 1 h after feeding than before feeding. This work constitutes a contribution to our knowledge of the physiology of crustaceans and attempts to clarify the possible mechanisms of digestive settings associated with hyper-regulation.

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