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Biomedical subjects

G Flik

Publications and source records attributed to G Flik.

At least 91 records · Page 5Linked to original sources
PubMed1990

Calcium absorption by fish intestine: the involvement of ATP- and sodium-dependent calcium extrusion mechanisms.

Measurements of unidirectional calcium fluxes in stripped intestinal epithelium of the tilapia, Oreochromis mossambicus, in the presence of ouabain or in the absence of sodium indicated that calcium absorption via the fish intestine is sodium dependent. Active Ca2+ transport mechanisms in the enterocyte plasma membrane were analyzed. The maximum capacity of the ATP-dependent Ca2+ pump (Vm: 0.63 nmol.min-1.mg-1, Km:27 nM Ca2+) is calculated to be 2.17 nmol.min-1.mg-1, correcting for 29% inside-out oriented vesicles in the membrane preparation. The maximum capacity of the Na+/Ca2+ exchanger with high affinity for Ca2+ (Vm:7.2 nmol.min-1.mg-1, Km:181 nM Ca2+) is calculated to be 13.6 nmol.min-1.mg-1, correcting for 53% resealed vesicles and assuming symmetrical behavior of the Na+/Ca2+ exchanger. The high affinity for Ca2+ and the sixfold higher capacity of the exchanger compared to the ATPase suggest strongly that the Na+/Ca2+ exchanger will contribute substantially to Ca2+ extrusion in the fish enterocyte. Further evidence for an important contribution of Na+/Ca2+ exchange to Ca2+ extrusion was obtained from studies in which the simultaneous operation of ATP- and Na(+)-gradient-driven Ca2+ pumps in inside-out vesicles was evaluated. The fish enterocyte appears to present a model for a Ca2+ transporting cell, in which Na+/Ca2+ exchange activity with high affinity for Ca2+ extrudes Ca2+ from the cell.

Adenosine Triphosphatases↗
PubMed1990

Prolactin cell activity and sodium balance in the acid-tolerant mudminnow Umbra pygmaea in acid and neutral water.

In East-American mudminnows, obtained from lakes and bogs of different water pH levels, prolactin cell size and ultrastructure reflect higher secretory activity in neutral water than in water of pH 3.5-6.5. This contrasts with observations on other species in which prolactin cell activity is higher at low water pH. Laboratory experiments involving acute exposure of mudminnows from pH 5.5 for 48 hr to different pH levels showed that prolactin secretion increased in water below pH 3 and above pH 6.5, which could be correlated with losses of blood electrolytes. No osmoregulatory stress was noticeable in the pH range of 3.5 to 5.5. Acclimation of fish for 5 to 6 months to either pH 4.5 or 7.2 confirmed that prolactin cell activity, as estimated with ultrastructural morphometry, was significantly higher in water of neutral pH than of pH 4.5. The growth rate was significantly higher at the lower pH. Determination of whole body sodium fluxes, with 24Na as a tracer, showed that both groups had a positive sodium balance. However, total body Na+ influx as well as efflux values were slightly but significantly reduced at pH 7.0 when compared to pH 4.5. The reduction of Na+ efflux at pH 7.0 is in line with increased secretion of prolactin since this hormone is known to limit the branchial permeability to water and ions, including Na+. The results show that the mudminnow is extremely acid tolerant, having an optimum water pH range of about 3.5 to 6.0, which is consistent with ecological observations. Prolactin secretion is at a minimum in this range. Exposure to neutral water represents osmoregulatory stress for the mudminnow, in contrast to all other teleost species examined so far.

Animals↗
PubMed1990

Rainbow trout corpuscles of Stannius: stanniocalcin synthesis in vitro.

In fresh-water rainbow trout, Oncorhynchus mykiss (formerly called Salmo gairdneri), experimentally induced mild hypercalcemia results in release of immunoreactive stanniocalcin from the corpuscles of Stannius (CS) and stimulated synthetic and releasing activities of the glands as measured in vitro. Pulse-chase experiments showed that stanniocalcin (STC) is a 56-kDa glycoprotein, processed from a 64-kDa precursor, prostanniocalcin (PSTC). PSTC and STC are homodimeric molecules that are readily split into monomers in the presence of reducing agents such as 2-mercaptoethanol. The monomeric form of PSTC and STC contains an approximately 5- to 6-kDa glycomoiety. Neither this sugar residue nor the NH2-terminal amino acid sequences of PSTC or STC proved to contain antigenic sites for the antiserum used in this study. Two-dimensional gel electrophoresis indicated the presence of several isoforms of PSTC and STC molecules that may reflect different stages of maturation of the (pro)hormone.

Animals↗
PubMed1989

Cadmium inhibition of the erythrocyte Ca2+ pump. A molecular interpretation.

The effects of cadmium (Cd2+) on transmembrane Ca2+ transport and on the membrane permeability for Ca2+ were studied in human erythrocytes. The erythrocyte Ca2+ pump is inhibited competitively by Cd2+ via interaction with the Ca2+ transport site of the carrier and not via interaction with its activator calmodulin. The affinity of the Ca2+ pump for Cd2+ is extremely high (KI = 2.0 nM Cd2+). Cd2+ (less than or equal to 10(-4) M) does not alter the membrane permeability for Ca2+. We conclude that the pivotal mechanism in the toxic action of Cd2+ is the inhibition of Ca2+-ATPase mediated Ca2+ extrusion. As a result Cd2+ disturbs intracellular Ca2+ homeostasis and may increase cytosolic Ca2+ (Ca2+i) to toxic levels.

Cadmium↗
PubMed1989

Immunocytochemical localization of hypocalcin in the endocrine cells of the corpuscles of Stannius in three teleost species (trout, flounder and goldfish).

In order to identify the cell-type responsible for the production of hypocalcin (the recently isolated hypocalcemic hormone of teleost fish), the corpuscles of Stannius (CS) of trout, flounder and goldfish, were immunocytochemically stained with antisera raised against trout hypocalcin. The secretory granules of the type-1 cells of the CS, considered to be the hypocalcin-producing cells, showed intense immunoreactivity in all species examined. However, in trout and flounder, the secretory granules produced by the type-2 cells, which have been suggested to represent a functionally different cell-type, also showed an intense immunoreactivity. In goldfish, no type-2 cells were observed. We tentatively conclude that type-1 and type-2 cells represent structurally different forms of the same functional cell-type.

Animals↗
PubMed1989

Immunocytochemical demonstration of a novel system of neuroendocrine peptidergic neurons in the pond snail Lymnaea stagnalis, with antisera to the teleostean hormone hypocalcin and mammalian parathyroid hormone.

Immunocytochemical staining with antisera raised against trout hypocalcin, the hypocalcemic hormone of the Stannius corpuscles and against bovine parathyroid hormone (bPTH1-84), revealed a new system of neuroendocrine neurons in the pond snail Lymnaea stagnalis. The neurons are located in small groups or single cells in the visceral, parietal, and pedal ganglia of the central nervous system. The axons of these cells are running to the periphery of the pleuroparietal, visceroparietal, and pleuropedal connections, the dorsopedal commissure, and to several nerves originating in the visceral, parietal, and pedal ganglia. The axons are ending with characteristic axonal distensions in the periphery of these connectives, commissure, and nerves. These regions probably act as neurohaemal areas. The affinity of this neuroendocrine system for both the anti-hypocalcin and anti-PTH sera is another indication for a special relationship between hypocalcin and PTH, which possess some immunological resemblance and similar biological activities, although no similarity in primary structure.

Animals↗
PubMed1989

Prolactin cell activity and sodium fluxes in tilapia (Oreochromis mossambicus) after long-term acclimation to acid water.

In tilapia exposed for 3 months to water of pH 4.5, prolactin cell activity, as estimated by ultrastructural morphometry and determination of prolactin synthesis in vitro, was significantly higher than in controls from neutral water. Sodium influx from the water was 50% lower than in the controls, indicating impaired branchial sodium uptake mechanisms. In contrast to predictions based on the results of short-term exposure to acid water--which is known to induce an increase of sodium efflux--the sodium efflux rate was reduced to 70% of the control value. It is concluded that tilapia are able to acclimate to acid water by successful control--probably via prolactin--of diffusional sodium losses across the integument, in particular the gill surface. This compensates for the impaired sodium uptake, and enables the fish to reestablish a positive sodium balance in acid water.

Acclimatization↗
PubMed1989

Calcitropic actions of prolactin in freshwater North American eel (Anguilla rostrata LeSueur).

In freshwater-acclimated American eels (Anguilla rostrata LeSueur), ovine prolactin and grafts of the part of the pituitary gland containing the prolactin cells induced hypercalcemia. The hypercalcemia was associated with increased uptake of calcium from the water (resulting from increased influx and decreased efflux) and with enhanced high-affinity Ca2+-adenosinetriphosphatase (ATPase) activity in the gills, the putative biochemical correlate of the branchial Ca2+ pump. Kinetic analyses of ATPase-mediated Ca2+ transport in plasma membrane vesicles of branchial epithelium provided evidence that prolactin enhanced the maximum velocity of the Ca2+ pump. Prolactin treatments raised plasma cortisol levels slightly but significantly in eels. However, cortisol per se was not hypercalcemic in eels and did not stimulate the branchial Ca2+ pump. We conclude that the hypercalcemic potency of prolactin in fish relates to its stimulatory action on active Ca2+ transport in the gills.

Adenosine Triphosphatases↗
PubMed1989

Ionic and total calcium levels in the blood of the European eel (Anguilla anguilla): effects of stanniectomy and hypocalcin replacement therapy.

Removal of the corpuscles of Stannius (STX) in the freshwater European eel causes a marked increase in the concentrations of blood ionic calcium and protein-bound calcium. The hypercalcaemia peaks 20 days after STX and lasts at least another 20 days. In stanniectomized eels hypocalcin decreased both blood ionic and total calcium concentrations. The reduction of plasma total calcium concentration by hypocalcin is attributed to a reduction in blood ionic calcium concentration. We conclude that hypocalcin regulates blood ionic calcium levels in fish.

Anguilla↗
PubMed1989

Rainbow trout hypocalcin stimulates bone resorption in embryonic mouse calvaria in vitro in a PTH-like fashion.

Hypocalcin, the major hormone with hypocalcaemic action in fish, was isolated from trout corpuscles of Stannius (SCs). The bioactivity of hypocalcin was assessed in a parathyroid hormone (PTH) bioassay involving bone resorption in embryonic mouse calvaria. Calcium and phosphate release and lactate production were stimulated in a dose-dependent manner by hypocalcin. On a molar basis about equal amounts of hypocalcin and PTH were required to obtain similar effects in this assay. Hypocalcin did not stimulate cyclic AMP production either in mouse calvaria or in cultured osteoblasts. In this respect hypocalcin resembles shortened or N-terminus-modified PTH molecules that induce bone resorption without increasing cyclic AMP levels. Since hypocalcin and PTH have comparable bioactivity in this mammalian bioassay (as well as in fish bioassays), we tentatively suggest that both hormones are structurally similar and that both hormones may act via the same receptors. The two hormones show no resemblance to one another in primary structure, so we suggest that they have similarities in tertiary structure.

Animals↗
PubMed1989

Cortisol stimulates whole body calcium uptake and the branchial calcium pump in freshwater rainbow trout.

Exposure of rainbow trout to a reduced ambient calcium level (from 490 to 25 mumol Ca2+/l) caused hypocalcaemia and induced a rapid increase (within 1 h) in systemic cortisol levels. Under conditions of low environmental calcium concentrations, cortisol levels remained increased for at least 8 days. After this time the in-vitro Ca2+-transport capacity of branchial basolateral membrane vesicles was increased due to stimulation of Ca2+-ATPase activity, presumably as a result of chloride cell proliferation. Pituitary prolactin cells were unaffected by low ambient calcium levels. Fish kept in water containing 490 mumol Ca2+/l and treated with cortisol for 7 days displayed an increase in whole body calcium uptake and an enhancement of the branchial calcium transport capacity; concomitantly, hypercalcaemia was observed. We conclude that, in the rainbow trout, cortisol exerts hypercalcaemic effects by stimulating Ca2+ uptake from the water and that this effect forms an intrinsic part of the established mineralocorticoid action of cortisol in fish.

Adenosine Triphosphatases↗
PubMed1988

Effects of epinephrine on branchial and renal calcium handling in the rainbow trout.

Acute exposure of rainbow trout (Salmo gairdneri) to low external calcium (25 microM) caused an immediate but transient increase in plasma epinephrine concentration that may have been related to a concomitant depression of blood pH. Intra-arterial infusion of epinephrine at normal ambient calcium levels (0.35 mM) for 4 h caused circulating levels of epinephrine to rise from 2.9 X 10(-9) to 8.0 X 10(-8) M but did not affect norepinephrine levels, or branchial unidirectional calcium fluxes. Active (ATP-dependent) calcium transport across basolateral plasma membranes prepared from gill epithelial cells was not affected by pretreatment of fish with epinephrine or by direct application of epinephrine or cAMP, in vitro. Epinephrine infusion elevated urine flow rate, decreased urine pH, and increased urine phosphate levels significantly. Net renal calcium efflux increased significantly as a result of the increased urine flow rate. It is concluded that epinephrine does not stimulate branchial calcium uptake or renal conservation of calcium in rainbow trout at normal external calcium levels and therefore we cautiously suggest that epinephrine is unlikely to be involved in calcium balance during periods of exposure to low external calcium. Instead, epinephrine may play a role in compensating the acid-base disturbances and the increased branchial water influx that are associated with exposure to low ambient calcium.

Animals↗
PubMed1988

Cadmium inhibits plasma membrane calcium transport.

The interaction of Cd2+ with the plasma membrane Ca2+-transporting ATPase of fish gills was studied. ATP-driven Ca2+-transport in basolateral membrane (BLM) vesicles was inhibited by Cd2+ with an I50 value of 3.0 nM at 0.25 microM free Ca2+, using EGTA, HEEDTA and NTA to buffer Ca2+ and Cd2+ concentrations. The inhibition was competitive in nature since the K0.5 value for Ca2+ increased linearly with increasing Cd2+ concentrations while the Vmax remained unchanged. The Ca2+ pump appeared to be calmodulin dependent, but we conclude that the inhibition by Cd2+ occurs directly on the Ca2+-binding site of the Ca2+-transporting ATPase and not via the Ca2+-binding sites of calmodulin. It is suggested that Cd2+-induced inhibition of Ca2+-transporting enzymes is the primary effect in the Cd2+ toxicity towards cells followed by several secondary effects due to a disturbed cellular Ca2+ metabolism. Our data illustrate that apparent stimulatory effects of low concentrations of Cd2+ on Ca2+-dependent enzymes may derive from increased free-Ca2+ levels when Cd2+ supersedes Ca2+ on the ligands.

Animals↗
PubMed1988

Identification of hypocalcin (teleocalcin) isolated from trout Stannius corpuscles.

We have isolated and purified a glycoprotein from the corpuscles of Stannius (CS) of trout, which we consider hypocalcin (also called teleocalcin), the major hypocalcemic hormone of fish. This product is present in relatively large amounts in the CS of several species (i.e., European eel, tilapia, goldfish, and carp). Hypocalcin is typically released from the CS in response to an experimentally induced increase of the blood calcium concentration. Ultrastructural observations show that after this treatment the type 1 cells, reportedly the hypocalcin-producing cell type of the CS, are almost completely degranulated. The isolated glycoprotein has an apparent molecular weight of 54 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This molecule appears susceptible to breakdown and is recovered upon concanavalin-A affinity chromatography as a 41 kDa product. Reducing agents such as mercaptoethanol or dithiothreitol employed, e.g., during standard electrophoretic techniques or during amino acid sequence analysis, allow only the recovery of 28 or 18 kDa products. Evidence is given that the 54 and 41 kDa products are dimer molecules, with the 28 and 18 kDa products as their respective monomeric constituents. The sequence of the first 33 N-terminal amino acids of these products and the composition of the sugar component are presented.

Amino Acid Sequence↗
PubMed1988

Control of prolactin secretion in the teleost Oreochromis mossambicus: effects of water acidification.

Prolactin secretion is stimulated markedly in fish exposed to water of pH 4. This phenomenon was used to study the control of prolactin secretion. Activation occurs irrespective of changes in plasma osmolarity or plasma sodium and calcium concentrations. After acute acidification of the water, which leads to a substantial fall in plasma osmolarity and plasma electrolyte levels, the activation of the prolactin cells is less marked than after gradual acidification of the water, when plasma osmolarity, plasma sodium, and plasma total and ionic calcium levels are not noticeably affected. When fish bearing an implanted rostral pars distalis of the pituitary gland are exposed to water of pH 4, both the in situ prolactin cells and the prolactin cells of the implant become activated only when the drop in water pH is acute and followed by a reduction in plasma osmolarity and electrolyte levels. When the rate of reduction of the pH is slow and not changing plasma osmolarity or sodium and calcium levels, the in situ prolactin cells are stimulated, but not those of the implants. We conclude that the activation of the prolactin cells in situ in fish in acid water is not mediated by reductions in plasma osmolarity, plasma sodium, total calcium, or ionic calcium, but by hypothalamic mechanisms. The drop in plasma osmolarity and electrolytes probably reflects excessive osmoregulatory stress and this may hamper instead of stimulate the response of the prolactin cells to osmoregulatory disturbance. The physiological significance of the in vitro activation of prolactin cells by reduced ambient osmolarity is unclear.

Acids↗
PubMed1988

Characterization of branchial transepithelial calcium fluxes in freshwater trout, Salmo gairdneri.

Experiments were performed to determine whether gill transepithelial calcium fluxes in the freshwater trout (Salmo gairdneri) are passive or require active transport and to characterize the mechanisms involved. A comparison of the in vivo unidirectional flux ratios with the flux ratios calculated according to the transepithelial electrochemical gradients revealed that calcium uptake from the water requires active transport of Ca2+. The inhibition of calcium uptake by external lanthanum, the specific deposition of lanthanum on the apical surface of chloride cells, and the favorable electrochemical gradient for calcium across the apical membrane suggest that the initial step in branchial calcium uptake is the passive entry of calcium into the cytosol of chloride cells through apical channels that are permeable to calcium. The study of gill basolateral plasma membrane vesicles demonstrated the existence of a high-affinity calmodulin-dependent calcium-transporting system [half-maximal Ca2+ concentration (K0.5) = 160 nM, Vmax = 1.86 nmol.min-1.mg protein-1]. This system actively transports calcium from the cytosol of chloride cells into the plasma against a sizeable electrochemical gradient, thereby completing the transepithelial uptake of calcium. Calcium efflux occurs passively through paracellular pathways between chloride cells and adjacent pavement cells or between neighboring pavement cells.

Animals↗
PubMed1988

Hypocalcin from Stannius corpuscles inhibits gill calcium uptake in trout.

Bidirectional whole body flux and branchial Ca2+ influx were measured in freshwater rainbow trout. Intra-arterial injections of homogenates of Stannius corpuscles (CS) as well as of a 54-kDa isolated product (hypocalcin) exerted an inhibitory effect on whole body Ca2+ influx, but did not effect Ca2+ efflux. Hypocalcin was more effective in reducing Ca2+ influx in trout acclimated to low-calcium freshwater than in fish from normal-calcium water. We conclude that the isolated product (hypocalcin) represents the hypocalcemic principle of the CS. Similar doses of hypocalcin caused quantitatively similar decreases in Ca2+ influx in vivo and in the isolated perfused head preparation. This indicates that the gills form the principle target for hypocalcin in trout. The branchial transepithelial potential did not change during hormone treatments. Possible mechanisms of hypocalcin action are suggested.

Animals↗