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PubMed2005

Actions of growth hormone on carbohydrate metabolism and osmoregulation of rainbow trout (Oncorhynchus mykiss).

Rainbow trout Oncorhynchus mykiss were injected intraperitoneally with slow-release implants of vegetable oil alone or containing ovine growth hormone (oGH) (2 and 5 microgg(-1) body weight), and sampled after 5 days to assess the simultaneous effects of GH on both osmoregulation and carbohydrate metabolism. An enhanced hypoosmoregulatory capacity of oGH-implanted fish is suggested by the increase observed in gill Na+,K+-ATPase activity, and the decrease observed in plasma ion concentration (Na+ and Cl-) and osmolality. GH treatment also elicited increased plasma glucose levels and metabolic changes in liver, gills, kidney, and brain. Major metabolic changes elicited by GH treatment included (1) decreased glycolytic potential and capacity for exporting glucose in liver, (2) enhanced glycogenolytic potential and capacity for use of exogenous glucose in gills and kidney, as well as increased glycolytic capacity in the later tissue, and (3) enhanced glycogenolytic and glycolytic capacities in brain. These metabolic changes elicited by GH treatment support a role for GH in the control of carbohydrate metabolism in salmonids that could be related either to the metabolic changes occurring during osmotic acclimation in nature (a process in which changes in GH levels and carbohydrate metabolism have both been reported) or to metabolic changes associated with growth.

Animals↗
PubMed2005

A perspective on the role of natriuretic peptides in amphibian osmoregulation.

The natriuretic peptide (NP) system is a complex family of peptides and receptors that is primarily linked to the maintenance of osmotic and cardiovascular homeostasis. In amphibians, the potential role(s) of NPs is complicated by the range of osmoregulatory strategies found in amphibians, and the different tissues that participate in osmoregulation. Atrial NP, brain NP, and C-type NP have been isolated or cloned from a number of species, which has enabled physiological studies to be performed with homologous peptides. In addition, three types of NP receptors have been cloned and partially characterised. Natriuretic peptides are always potent vasodilators in amphibian blood vessels, and ANP has been shown to increase the permeability of the microcirculation. In the perfused kidney, ANP causes vasodilation, diuresis and natriuresis that are caused by an increased GFR rather than effects in the renal tubules. These data are supported by the presence of ANP receptors only on the glomeruli and renal blood vessels. In the bladder and skin, the function of NPs is enigmatic because physiological analysis of the effects of ANP on bladder and skin function has yielded conflicting data with no clear role for NPs being revealed. Overall, NPs often have no direct effect, but in some studies they have been shown to inhibit the function of AVT. In addition, there is evidence that ANP can inhibit salt retention in amphibians since it can inhibit the ability of adrenocorticotrophic hormone or angiotensin II to stimulate corticosteroid secretion. It is proposed that an important role for cardiac NPs could be in the control of hypervolaemia during periods of rapid rehydration, which occurs in terrestrial amphibians.

Amino Acid Sequence↗
PubMed2006

Effects of acute stress on osmoregulation, feed intake, IGF-1, and cortisol in yearling steelhead trout (Oncorhynchus mykiss) during seawater adaptation.

Juvenile steelhead (Oncorhynchus mykiss) acclimated to freshwater (FW) were subjected for 3 h to confinement stress in FW, and subsequently saltwater (SW, 25 ppt) was introduced to all tanks. Fish were sampled immediately after the stress treatment, and 1, 7, and 14 days after introduction of SW. Electrolytes, cortisol, glucose, and lactate showed the typical stress response expected after stress treatment in FW. Fish regained osmotic balance within 24 h. Glucose concentrations were increasing throughout the experiment and lactate levels stayed elevated during the time spent in SW. Insulin-like growth factor-1 (IGF-1) did not show an immediate response to stress but after transfer to SW we detected significantly higher concentrations for control fish at days 1 and 14. The differences in IGF-1 levels between stressed and control fish are not reflected in SW adaptability but positive correlations between IGF-1 and electrolyte levels in control fish may indicate its role for osmoregulation. Confinement stress did not impair feed intake subsequently in SW, but our results suggest that feed intake was suppressed by the change of the media from FW to SW.

Adaptation, Physiological↗
PubMed1998

Methylmercury-induced inhibition of regulatory volume decrease in astrocytes: characterization of osmoregulator efflux and its reversal by amiloride.

Swelling of neonatal rat primary astrocyte cultures by hypotonic media leads to regulatory volume decrease (RVD) and the resumption of resting cell volume. RVD is associated with activation of conductive K+ and Cl- channels, allowing for the escape of KCl, as well as the release of osmoregulators, such as taurine and myoinositol. As we have previously shown [D. Vitarella, H.K. Kimelberg, M. Aschner, Inhibition of RVD in swollen rat primary astrocyte cultures by methylmercury (MeHg) is due to increase amiloride-sensitive Na+ uptake, Brain Res. 732 (1996) 169-178.], MeHg, when added to hypotonic buffer inhibits RVD, primarily due to increased cellular permeability to Na+ via the Na+/H+ antiporter. The present study was, therefore, undertaken to assess the ability of cation-anion cotransport blockers to reverse the inhibitory effect of MeHg on RVD in swollen astrocytes, and to further characterize MeHg-induced changes in astrocytic osmoregulatory release processes. The studies demonstrate the following: (1) MeHg-induced inhibition of RVD is partially inhibited by the Na+/H+ antiporter blocker, amiloride, but not SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid), DIDS (4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid), furosemide or bumetanide; (2) exposure of swollen astrocytes to MeHg is associated with specific effects on osmoregulatory release, leading to significant inhibition of taurine release and a significant increase in potassium and myoinositol release compared with release in hypotonic conditions.

Amiloride↗
PubMed1998

The Na+ and K+ transport deficiency of an E. coli mutant lacking the NhaA and NhaB proteins is apparent and caused by impaired osmoregulation.

Cells of the E. coli mutant EP432, which lacks the two Na+/H+ antiporters, NhaA and NhaB, have been reported to have an impaired sodium transport activity (Harel-Bronstein et al. (1995) J. Biol. Chem. 270, 3816-3822). Here we report that active transport of Na+ in EP432 cells can be restored to wild-type levels, either by a high K+ concentration or by an increase in the medium osmolarity. We suggest that this mutant is primarily deficient in osmoregulation rather than in cation transport per se.

Bacterial Proteins↗
PubMed1989

Smoltification and seawater adaptation in coho salmon (Oncorhynchus kisutch): plasma calcium regulation, osmoregulation, and calcitonin.

In order to examine the dynamics of ion regulation, osmoregulation, and plasma calcitonin during the parr-smolt transformation (smoltification), blood and gill tissue were collected from yearling coho salmon, Oncorhynchus kisutch, from February to October. Fish were kept in fresh water (FW) throughout this period. In addition, fish were exposed to seawater (SW) at the peak of smoltification in mid-April, and samples from these fish were collected until July. Plasma osmolality, gill Na+,K+-ATPase activity, plasma levels of calcitonin, and free and total calcium and magnesium were measured. SW adaptability of FW fish was assessed throughout the study by measurements of plasma osmolality following a 24-hr exposure to seawater. The greatest hypoosmoregulatory ability occurred in April-May, although SW-adapted fish had higher plasma osmolality than FW-adapted fish at all times. Gill Na+,K+-ATPase activity in FW-adapted fish increased from April to June and increased rapidly following exposure of fish to SW, and remained elevated in SW-adapted fish. Free plasma calcium and magnesium levels increased following SW exposure, but returned to prior levels within 1 week. Netting and confinement stress during sampling caused an increase in plasma osmolality and free calcium and magnesium levels in both FW- and SW-adapted fish. Changes in hypoosmoregulatory ability during smoltification and SW adaptation were correlated with changes in gill Na+,K+-ATPase activity. A sharp transitory peak in plasma calcitonin levels occurred early in smoltification (March) and in SW-adapted fish in June. Plasma calcitonin levels gradually increased in FW-adapted fish during the period of desmoltification. However, no change in plasma calcitonin levels occurred during SW-induced hypercalcemia, suggesting that the hormone does not play a major role in short-term plasma calcium regulation in coho salmon.

Acclimatization↗
PubMed1983

Intestinal osmoregulation of single component meals in neonatal piglets.

The ability of the neonatal stomach and duodenum to modify the osmolality of hypotonic and hypertonic meals containing a single nutrient was studied in 3- to 5-day-old piglets. Test meals were administered through a gastrostomy and samples of gastric content and all proximal jejunal effluent were collected every 30 minutes for 2 hours. Hypotonic meals (149 mosm/L) and hypertonic meals (497 mosm/L) of glucose, crystalline amino acids, or lipids were used. Results were similar after all meals. There were significant changes in intragastric osmolality at each postprandial time period. However, osmolality never returned to basal levels. During duodenal transit there were further increases in osmolality after hypotonic meals and decreases after hypertonic meals but isotonic levels were never achieved. As a result, chyme of widely divergent osmolality entered the jejunum until late in the observation periods. Influx of sodium, potassium, and chloride were similar after all meals and were independent of original meal osmolality. These data indicate that, in awake, unstressed neonatal piglets, the stomach and duodenum are capable of effective but incomplete osmoregulation of hypotonic and hypertonic single component meals.

Animals↗
PubMed1983

Chronic hypernatremia from a congenital defect in osmoregulation of thirst and vasopressin.

An infant with microcephaly and delayed development was found to have chronic asymptomatic hypernatremia. Computerized brain tomography disclosed dysplasia of the midline structures, septum pellucidum and corpus collosum. Evaluation revealed defective osmoregulation, hypothalamic hypothyroidism, and hypogonadotropinism. He showed no desire to drink at plasma osmolalities over 330 mOsm/kg. His plasma vasopressin levels (less than or equal to 1.4 pg/ml) were inappropriately low relative to his high levels of plasma osmolality (greater than or equal to 310 mOsm/kg), which might be accounted for by either deficient neurohypophyseal vasopressin stores or disturbance of the hypothalamic osmoreceptors governing vasopressin. The first possibility was ruled out by demonstrating normal vasopressin response (167 pg/ml) to nonosmotic (emetic) stimulation. Under baseline conditions, his urine was concentrated up to 747 mOsm/kg and urine volume was low. With water loading, maximal water diuresis developed (urine osmolality 68 mOsm/kg), but his plasma osmolality remained in the hyperosmolar range (312 mOsm/kg). Treatment with a vasopressin analogue, desamino-D-arginine vasopressin, and forced hydration restored plasma osmolality and plasma sodium to normal. These findings indicate a severe defect in the hypothalamic osmoreceptors controlling thirst and vasopressin secretion with normal vasopressin stores and preserved vasopressin responsiveness to nonosmotic stimuli. To our knowledge, this report provides the first documentation of selective osmoreceptor defect in conjunction with congenital dysplasia of midline brain structures.

Brain↗
PubMed2001

Osmoregulation and cell volume regulation in the preimplantation embryo.

The early preimplantation mammalian embryo possesses mechanisms that regulate intracellular osmolarity and cell volume. While transport of osmotically active inorganic ions might play a role in this process in embryos, the major mechanisms that have been identified and studied are those that employ organic osmolytes. Organic osmolytes provide a substantial portion of intracellular osmotic support in embryos and are required for their development under in vivo conditions. The main osmolytes that have been identified in cleavage stage embryos are accumulated via two transport systems of the neurotransmitter transporter family active in early preimplantation embryos--the glycine transport system (GLY) and the beta-amino acid transport system (system beta). While system beta has been established to have a similar role in many other cells, this is a novel function for the GLY transport system. The intracellular concentration of organic osmolytes such as glycine in early preimplantation embryos is regulated by tonicity, allowing the embryo to regulate its volume against shrinkage and to control its internal osmolarity. In addition, the cells of the embryo can regulate against an increase in volume via controlled release of osmolytes from the cytoplasm. This is mediated by a swelling-activated anion channel that is also highly permeable to a range of organic osmolytes, and which closely resembles similar channels found in many other cell types (VSOAC channels). Together, these mechanisms appear to regulate cell volume in the egg through the early cleavage stages of embryogenesis, after which there are indications that the mechanisms of osmoregulation change.

Amino Acids↗
PubMed1998

Osmoregulation of vasopressin neurons: a synergy of intrinsic and synaptic processes.

The release of vasopressin into the general circulation varies as a function of plasma osmolality and therefore plays a major role in systemic osmoregulation. In vivo, the secretion of this hormone in the neurohypophysis is primarily determined by the rate of action potential discharge of the magnocellular neurosecretory cells (MNCs) in the hypothalamus. Experiments done over the past 20 years have clarified much of the neurophysiological basis underlying this important osmoregulatory reflex. As discussed here, recent findings indicate that the regulation of the firing rate of MNCs during changes in systemic osmolality involves the concerted modulation of mechanosensitive ion channels in MNCs, as well as excitatory glutamatergic inputs derived from forebrain regions such as the organum vasculosum of the lamina terminalis.

Action Potentials↗
PubMed2002

Osmoregulation and its importance to food-borne microorganisms.

The control of water activity has been used as a means of preserving foods for thousands of years. This preservation strategy presents food-borne microorganisms with serious problems, many of which relate to the management of water flow. Although the specific details of how each organism deals with these problems are different, several common themes have emerged. Bacteria induce specific responses. both physiological and genetic, to respond to either the loss or the gain of water, triggered by changes in the osmolarity of the environment. Many of the key systems have now been identified and the mechanisms of their regulation are beginning to be understood. Here we review recent developments in the field of bacterial osmoregulation with emphasis on key food-borne genera.

Bacterial Physiological Phenomena↗
PubMed2000

PLGamide characterization and role in osmoregulation in leech brain.

Neurons immunoreactive to an antiserum specifically directed against the Prolyl-Leucyl-Glycinamide peptide (PLGamide: Melanocyte Inhibiting Factor: MIF) were detected in the brain of the leech Theromyzon tessulatum. Radioimmunoassay titrations of the PLGamide-like material at different physiological stages of the life cycle indicated a maximal amount at stage 3B, which is correlated to phase of both maximal water uptake and coelomic vitellogenin accumulation. In vivo experiments demonstrate that this cerebral PLGamide-like material is an anti-diuretic factor that would act at stage 3B in order to permit a water uptake leading to water retention allowing coelomic yolk protein accumulation. In brains of the Gnatobdellid leech Hirudo medicinalis and the Pharyngobdellid leech Erpobdella octoculata, anti-PLGamide material was also detected with an amount not differing with the degree of sex maturation of the animals, confirming the link between osmoregulation and ovogenesis in rhynchobdellid leeches. Using a combination of biochemical techniques including high-pressure gel permeation chromatography followed by reversed-phase HPLC on brain extracts and Edman degradation, we demonstrated the presence of an authentic MIF-1 peptide in leech brain. Finally, since in vertebrates MIF-1 belongs to the non-classical opioid peptide family, we studied its binding displacement, in contrast to morphine, on mu-receptors and on nitric oxide (NO) release experiments in leech brain. PLGamide did not bind to mu-alkaloid opioid receptors and did not stimulate NO release.

Animals↗
PubMed2001

Betaine aids in the osmoregulation of duodenal epithelium of broiler chicks, and affects the movement of water across the small intestinal epithelium in vitro.

In Experiment 1, the water holding capacity of broiler chick intestinal tissue was studied in vitro. The chicks were fed with corn-based diets with or without a 0.2% betaine supplementation in the drinking water. Slices from duodenum and jejunum were incubated in iso-osmotic (300 mM) or hyperosmotic saline (600 mM) with or without 10 mM betaine. The water volume of tissue slices was studied by adding tritiated water in the incubation medium while [14C]inulin was used to correct for the adherent water. After 30 min of incubation, by which time the steady-state of tritium influx had been achieved, the 3H and 14C-activities of the tissue slices were measured. The ileal and duodenal tissues incubated in the hyperosmotic saline accumulated less tritium than those incubated in iso-osmotic saline. Duodenal slices incubated in hyperosmotic saline with the presence of betaine showed a tritium content similar to slices incubated in iso-osmotic saline. The data suggest that the presence of betaine helped the duodenal, but not jejunal, epithelium to maintain water balance in hyperosmotic conditions. The dietary betaine supplementation diminished the differences between the incubation treatments in duodenal, but not in ileal tissue. In Experiment 2, the same double labeling method, but with shorter incubation times, was used to assess the rate of water flux from the incubation medium to duodenal or jejunal slices. The dietary treatments (as in Experiment 1) had little effect on the results. Betaine in the hyperosmotic saline significantly decreased the rate of tritium accumulation into the tissue slices, indicating that betaine slowed down the influx of water to the epithelium. We suggest that betaine affects the movement of water across the intestinal epithelium and has a role in the osmoregulation of small intestine of broiler chicks.

Animals↗
PubMed2001

Osmoregulation in Onymacris rugatipennis, a free-ranging tenebrionid beetle from the Namib Desert.

Haemolymph levels of organic and inorganic constituents were investigated in the tenebrionid Onymacris rugatipennis during dehydration and rehydration. The major osmolar effectors are sodium (26%), chloride (24%), amino acids (18%), and sugars (11%); regulation of haemolymph osmotic pressure (OP) during dehydration is effected largely by a reduction in the haemolymph content of these constituents. Changes in amino acid levels are not the result of interchanges with soluble protein. During rehydration, the main contributors to osmoregulation are sodium (26%), chloride (24%), and an increase in haemolymph solute(s) not measured in this study (31%). Of the sodium removed from the haemolymph during dehydration, 21.2% was excreted. Faecal losses of potassium during dehydration far exceeded the amounts removed from the haemolymph; however, haemolymph potassium levels were strongly regulated during rehydration. Regulatory efficiency increases as desiccation proceeds, and is greatest only when this species is severely challenged.

Amino Acids↗
PubMed2001

Molecular physiology of osmoregulation in eels and other teleosts: the role of transporter isoforms and gene duplication.

This review focuses on recent developments in the molecular biology of ion and water transporter genes in fish and the potential role of their products in osmoregulation in both freshwater and seawater environments. In particular details of isoforms of various ATPases, co-transporters, exchangers and ion channels in the eel as well as other teleost species are described. Many of the teleost transporter isoforms discovered so far, appear to occur as twin or duplicate copies compared to their homologous counterparts in higher vertebrates, although these duplicate isoforms often have distinct tissue-specific and developmental stage-dependent expression patterns. The possible meaning of this information will be examined in relation to the fish genome duplication debate.

Amino Acid Sequence↗
PubMed2003

The effect of copper on osmoregulation in the freshwater amphipod Gammarus pulex.

The influence of copper on osmoregulation in the freshwater amphipod Gammarus pulex was determined from the analysis of water permeability, haemolymph sodium concentration, sodium influx and gill Na(+)/K(+) ATPase and Mg(2+) ATPase activity. Exposure to nominal copper concentrations of 100 microg l(-1) or greater caused a significant reduction in both haemolymph sodium concentration and sodium influx within 4 h. Measurements of water permeability, expressed as the half-time of exchange of body water (t(1/2)), excluded structural gill damage as the cause of this fall in haemolymph sodium. Copper at 10 microg l(-1) or above in the assay solution significantly reduced gill Na(+)/K(+) ATPase activity. In contrast gill Mg(2+) ATPase activity was markedly less affected by copper. These differences in enzyme sensitivity were considered with respect to the potential mechanisms of copper toxicity.

Amphipoda↗
PubMed2002

Probing the nucleotide binding domain of the osmoregulator EnvZ using fluorescent nucleotide derivatives.

EnvZ is a histidine protein kinase important for osmoregulation in bacteria. While structural data are available for this enzyme, the nucleotide binding pocket is not well characterized. The ATP binding domain (EnvZB) was expressed, and its ability to bind nucleotide derivatives was assessed using equilbrium and stopped-flow fluorescence spectroscopy. The fluorescence emission of the trinitrophenyl derivatives, TNP-ATP and TNP-ADP, increase upon binding to EnvZB. The fluorescence enhancements were quantitatively abolished in the presence of excess ADP, indicating that the fluorescent probes occupy the nucleotide binding pocket. Both TNP-ATP and TNP-ADP bind to EnvZB with high affinity (K(d) = 2-3 microM). The TNP moiety attached to the ribose ring does not impede access of the fluorescent nucleotide into the binding pocket. The association rate constant for TNP-ADP is 7 microM(-1) s(-1), a value consistent with those for natural nucleotides and the eucaryotic protein kinases. Using competition experiments, it was found that ATP and ADP bind 30- and 150-fold more poorly, respectively, than the corresponding TNP-derivatized forms. Surprisingly, the physiological metal Mg(2+) is not required for ADP binding and only enhances ATP affinity by 3-fold. Although portions of the nucleotide pocket are disordered, the recombinant enzyme is highly stable, unfolding only at temperatures in excess of 70 degrees C. The unusually high affinity of the TNP derivatives compared to the natural nucleotides suggests that hydrophobic substitutions on the ribose ring enforce an altered binding mode that may be exploited for drug design strategies.

Amino Acid Sequence↗
PubMed2005

A structural model for the osmosensor, transporter, and osmoregulator ProP of Escherichia coli.

Transporter ProP of Escherichia coli, a member of the major facilitator superfamily (MFS), acts as an osmosensor and an osmoregulator in cells and after purification and reconstitution in proteoliposomes. H(+)-osmoprotectant symport via ProP is activated when medium osmolality is elevated with membrane impermeant osmolytes. The three-dimensional structure of ProP was modeled with the crystal structure of MFS member GlpT as a template. This GlpT structure represents the inward (or cytoplasm)-facing conformation predicted by the alternating access model for transport. LacZ-PhoA fusion analysis and site-directed fluorescence labeling substantiated the membrane topology and orientation predicted by this model and most hydropathy analyses. The model predicts the presence of a proton pathway within the N-terminal six-helix bundle of ProP (as opposed to the corresponding pathway found within the C-terminal helix bundle of its paralogue, LacY). Replacement of residues within the N-terminal helix bundle impaired the osmotic activation of ProP, providing the first indication that residues outside the C-terminal domain are involved in osmosensing. Some residues that were accessible from the periplasmic side, as predicted by the structural model, were more susceptible to covalent labeling in permeabilized membrane fractions than in intact bacteria. These residues may be accessible from the cytoplasmic side in structures not represented by our current model, or their limited exposure in vivo may reflect constraints on transporter structure that are related to its osmosensory mechanism.

Alkaline Phosphatase↗