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Taurine and osmoregulation. II. Administration of taurine analogues affords cerebral osmoprotection during chronic hypernatremic dehydration.

We have previously shown that in the cat, taurine is an osmoprotective molecule that lessens mortality, neurological morbidity, and brain-cell dehydration during chronic hypernatremic dehydration. We examined the ability of two taurine analogues to afford cerebral osmoprotection in rats. Pretreatment with guanidinoethane sulfonate, a competitive antagonist for beta-amino acid transport, as a 1% drinking solution for ten days led to a significant reduction in brain-cell dehydration. Thus, total brain-cell water was higher in experimental vs control animals (544.3 +/- 36.8 vs 478.2 +/- 12.7 mL/100 g of fat-free dry solids [FFDS]) and the difference was almost exclusively derived from the intracellular water compartment (452.7 +/- 27.3 vs 371.4 +/- 7.7 mL/100 g of FFDS). Pretreatment with taltrimide, a lipophilic taurine derivative (intraperitoneal injection of 200 mg/kg for four days), led to similar results. Total brain-tissue water was significantly higher in experimental vs control rats (507.6 +/- 18.8 vs 363.2 +/- 9.5 mL/100 g of FFDS), with the difference primarily derived from the intracellular water space (372.8 +/- 18.1 vs 221.3 +/- 13.1 mL/100 g of FFDS). These results suggest that the cerebral response to chronic hypertonic stress includes accelerated transmembrane flux of osmoprotective solutes in addition to mobilization from sequestered intracellular storage sites in an attempt to increase the cytosolic pool of osmotically active molecules.

Animals↗

Osmoregulation and salinity effects on the expression and activity of Na+,K(+)-ATPase in the gills of European sea bass, Dicentrarchus labrax (L.).

The European sea bass, Dicentrarchus labrax, tolerates salinities ranging from freshwater (FW) to hypersaline conditions. In two experiments, we analysed changes in plasma ions, muscle water content (MWC), gill Na+,K(+)-ATPase activity, and alpha-subunit mRNA expression during the course of acclimation from 15 ppt salt water to FW or high salinity seawater (HSSW). In Experiment 1, fish (6.2 +/- 1.1 g) were acclimated from 15 ppt to either FW, 5, 15, 25, 50, or 60 ppt SW and sampled after 10 days. Gill Na+,K(+)-ATPase activity was stimulated in FW- and in 50 and 60 ppt SW-groups relative to the 15 ppt control group. In Experiment 2, subgroups of fish (89 +/- 7 g) were transferred from 15 ppt SW to FW or 50 ppt SW, and sampled 1, 2, 4, and 10 days later. Plasma osmolality, [Na+] and [Cl-] decreased in the FW-group and increased in the HSSW-group one day after transfer and lasting until day 10. This was accompanied by a pronounced increase in MWC in the FW-group and an insignificant decrease in the HSSW-group. The plasma [Na+]:[Cl-]-ratio increased markedly in the FW-group and decreased slightly in the HSSW-group, suggesting acid-base balance disturbances after transfer. Gill Na+,K(+)-ATPase activity was unchanged in 15 ppt SW but doubled in FW- and HSSW-groups after transfer. In both groups, this was preceded by a 2- to 5-fold elevation of the gill alpha-subunit Na+,K(+)-ATPase mRNA level. Thus increased expression of alpha-subunit mRNA is part of the molecular mechanism of both FW and SW acclimation in sea bass. Gill Na+,K(+)-ATPase Na(+)-, K(+)-, and ouabain-affinity were similar in fish acclimated to FW, 15 ppt, and HSSW, suggesting that identical isoforms of the catalytic subunit of the enzyme are expressed irrespective of salinity.

Adaptation, Physiological↗

Possible functions of oxytocin/vasopressin-superfamily peptides in annelids with special reference to reproduction and osmoregulation.

Annetocin is an earthworm oxytocin-related peptide that we previously isolated from the whole body of a lumbricid earthworm Eisenia foetida. We have reported that annetocin induces egg-laying-like behaviors in E. foetida and a gnathobdellid leech, Whitmania pigra, when it is injected into the respective animals. The present study was undertaken to probe physiological functions of invertebrate oxytocin-vasopressin-superfamily peptides with special reference to reproductive and osmoregulatory events in which vertebrate peptides of this superfamily are involved. Annetocin, Lys-conopressin (a leech vasopressin-related peptide) and two analog peptides, [Tyr(3)]-annetocin ((3)Y-annetocin) and [Phe(3)]-annetocin ((3)F-annetocin), were compared for their activities to induce egg-laying-like behavior and to change body weight as a measure of water balance in the leech W. pigra. Injection of annetocin, Lys-conopressin, and (3)F-annetocin caused both egg-laying-like behavior and reduction of body weight in the animals, but (3)Y-annetocin induced neither. Furthermore, leeches in the non-breeding season responded to peptides less conspicuously than those in the breeding season. Such a concomitant induction of egg-laying-like behavior and body-weight reduction suggests that these two phenomena are unitary and might be accounted for by the fact that egg-laying in leeches and earthworms is accompanied by secretion of a large quantity of mucus, which should significantly contribute to body-weight loss. J. Exp. Zool. 284:401-406, 1999.

Amino Acid Sequence↗

DNA twist, flexibility and transcription of the osmoregulated proU promoter of Salmonella typhimurium.

Transcription from many bacterial promoters is sensitive to the level of DNA supercoiling. We have investigated the mechanism by which environmentally induced changes in DNA supercoiling might regulate transcription. For the proU promoter of Salmonella typhimurium, osmotically induced changes in DNA topology appear to play a primary regulatory role. Changes in DNA supercoiling (linking number; delta Lk) are partitioned into changes in the winding of the strands of the double helix about themselves (twist; delta Tw) and/or elastic deformations or flexibility of the DNA helix (writhe; delta Wr). Mutations of the proU promoter were isolated in vivo, or generated in vitro, which altered the spacing between the -10 and -35 motifs. Studies on these mutant promoters, both in vivo and in vitro, exclude models in which changes in DNA twist play a regulatory role. Instead, our data suggest that increased DNA flexibility, reflecting the osmotically induced increase in negative supercoiling of DNA, is required for promoter activation.

Amino Acid Transport Systems↗

A preliminary investigation into the roles played by the rectal gland and kidneys in the osmoregulation of the striped dogfish Poroderma africanum.

Presented here are the results of a preliminary investigation into ionic and osmotic regulation by the kidneys and rectal gland of the striped dogfish, Poroderma africanum. Fish with ligated rectal glands showed an increase in blood concentration of sodium and chloride within a short time period, reaching a maximum after four days. The blood concentration of the two ions then decreased over the following ten days. Control animals showed relatively unchanged blood-sodium and chloride levels, over the entire 14-day period. After salt loading, both control animals and those with ligated rectal glands showed initial rise in blood sodium and chloride levels, but these returned towards initial values within seven hours of injection. Comparison of the two groups indicates that the rectal gland may control blood-chloride levels more so than -sodium, although its action as a salt regulator does not seem very pronounced in either case. Urine and rectal gland fluid, were collected as a compound fluid, from normal fish, and the estimated cloacal salt loss is discussed. Urine from normal fish was also collected separately and was analysed for its contribution to salt loss. Results are discussed and compared with previous relevant findings.

Animals↗

Embryonic osmoregulation: consequences of high and low water loss during incubation of the chicken egg.

The rates of water loss of domestic chicken eggs were varied during incubation to measure the osmoregulatory ability of the avian embryo. Egg water loss was increased by drilling holes in the eggshell over the airspace on day 13 (I = 21 days) and then placing these eggs in a low relative humidity (r.h.: 0-10%) incubator until hatch. Egg water loss was decreased by placing other eggs in a high-r.h. (85-90%) incubator on day 0. Eggs with low water loss (approximately 6% of initial fresh mass [IFM]) produced embryos and yolks that were not different in wet or dry mass when compared to control eggs that lost approximately 12% of IFM. However, 1-4 gm of excess albumen were left in low-water-loss eggs on day 21. Hatching success was 71% and 89% for low and control eggs, respectively. Low egg water loss did not appear to disturb embryonic growth. The allantoic fluid volume and millimolar allantoic Na+ and Cl- ions declined faster with high and slower with low rates of water loss. Thus, excess water was lost as a result of increased movement of water out of allantoic fluid, which was due to increased active transport of Na+ ions by the chorioallantoic membrane (CAM). Eggs with high water loss had elevated Cl- levels after day 17 in plasma and amniotic fluid, which indicated a period of osmotic stress after depletion of allantoic fluid between day 18 and hatch. The decrease in wet embryo mass measured in embryos from high-water-loss eggs was due principally to dehydration of skin. Embryonic skin may serve as an emergency water reservoir during osmotic stress. Dehydrated chicks produced from high-water-loss eggs were 6 gm less in wet mass at hatch compared to controls. However, these chicks regained the water deficit 7 days after hatch and grew at a rate not different from control chicks through 6 weeks of age. Total egg water loss of 12% of IFM results in highest hatching success. However, water losses between 6% and 20% of IFM do not appear to affect adversely the growth or water content of the chick. Water losses above 20% of IFM cause early depletion of allantoic fluid, prolong the period of osmotic stress, and result in subsequent dehydration of blood, amniotic fluid, and embryonic skin.(ABSTRACT TRUNCATED AT 400 WORDS)

Allantois↗

Energetics of osmoregulation: I. Oxygen consumption by Fundulus heteroclitus.

We have developed a flow-through method for measuring oxygen consumption in fish which allows continuous monitoring over periods of days with good accuracy. Our goal was to determine the changes in basal metabolic rate in estuarine fish as a function of salinity. We show that in Fundulus heteroclitus, the oxygen consumption drops by 50% during the first 12 hr in the respirometer, as the fish cease exploratory movements. We have determined the influence of temperature and body size on resting respiratory rate, but failed to find any circadian or tidal rhythm in aerobic respiration. With these variables controlled, we determined that changing from 10 to 30 ppt water had no demonstrable effect on oxygen uptake. Since there must be a large change in osmotic flux due to this change in salinity, it appears that the fish might be diverting energy from other uses rather than increasing aerobic energy production to meet the increased osmoregulatory work load.

Animals↗

Osmoreceptors, osmoreception, and osmoregulation.

Mammals have evolved sophisticated behavioral and physiological responses to oppose changes in the osmolality of their extracellular fluid. The behavioral approach consists of regulating the intake of salt and water through changes in sodium appetite and thirst. The physiological approach comprises adjustments of renal excretion of water and sodium which are achieved through changes in the release of antidiuretic and natriuretic hormones. Individually, these osmoregulatory responses are controlled by "osmoreceptors": groups of specialized nerve cells capable of transducing changes in external osmotic pressure into meaningful electrical signals. Some of these sensors are located in the region of the hepatic portal vein, a strategic site allowing early detection of the osmotic impact of ingested foods and fluids. Changes in systemic osmolality, however, are detected centrally, within regions that include the medial preoptic area, the median preoptic nucleus, the organum vasculosum lamina terminalis (OVLT), the subfornical organ, and the supraoptic nucleus (SON). While studies have indicated that these central and peripheral osmoreceptors participate in the control of osmoregulatory responses, little is known of the mechanisms by which this is achieved. One notable exception, however, consists of the osmotic control of electrical activity in SON neurons which, in the rat, contributes to the regulation of natriuresis and diuresis through effects on the secretion of oxytocin and vasopressin. Previous studies have shown that these cells are respectively excited and inhibited by hypertonic and hypotonic conditions. Experiments in vitro indicate that these responses result from both the endogenous osmosensitivity of these cells and changes in synaptic drive. Patch-clamp analysis has revealed that SON neurons are respectively depolarized and hyperpolarized by increases and decreases in external osmolality and that these intrinsic responses result from changes in the activity of mechanosensitive cationic channels. Moreover, intracellular recordings in hypothalamic explants have shown that changes in electrical activity are associated with proportional changes in the frequency of glutamatergic excitatory postsynaptic potentials derived from osmosensitive OVLT neurons. Both of these mechanisms, therefore, may participate in the osmotic regulation of neurohypophysial hormone release in situ.

Animals↗

Seasonal changes in osmoregulation, cortisol, and cortisol receptor activity in the gills of parr/smolt of steelhead trout and steelhead-rainbow trout hybrids, Oncorhynchus mykiss.

Steelhead and hybrid trout (rainbow trout male, steelhead female) presmolts and smolts, Oncorhynchus mykiss, exposed to 30/1000 seawater (SW) for 24 hr manifested progressively lower plasma sodium values from December to May. The increase in plasma sodium after 24 hr in SW diminished significantly after March, with the hybrids (which were heavier in weight than the steelheads) reaching their peak osmoregulatory ability in the middle of April, and the steelheads in late May. In fresh water there was little difference in plasma osmotic concentration but steelhead plasma chloride fell significantly from March 30 levels to May 23 levels, whereas hybrid plasma chloride levels did not change during this time. In steelheads but not hybrids, plasma cortisol correlated negatively with plasma sodium of fish exposed 24 hr in SW. However, in both steelheads and hybrids, plasma sodium values 24 hr after SW exposure had an inverse correlation with cytosol cortisol from the gills. Again, in both steelheads and hybrids, plasma values of fish exposed 24 hr in SW correlated positively with cytosol receptor activity. In hybrids, the nuclear extract receptor activity, Nmax, increased significantly until late March and then dropped significantly thereafter. In steelheads, there were no significant differences but Nmax followed the same trends as occurred in hybrids. Thus seasonal changes in cortisol and cortisol receptors in steelhead occur during smoltification and preparation for marine life and suggest an important role for cortisol. In hybrids there appears to be a reduction in seasonality.

Animals↗

Hormonal control of osmoregulation in the channel catfish Ictalurus punctatus.

Prolactin (PRL) is an important hormone for freshwater adaptation in many teleost species. In some euryhaline fishes, growth hormone (GH) and cortisol are involved in seawater adaptation by stimulating ion extrusion. When channel catfish (Ictalurus punctatus) were transferred from fresh water to dilute seawater (300-400 mOsm), their plasma osmolality was always higher than the environmental salinity. In correlation with the increase in plasma osmolality, significant increases in plasma cortisol were observed. However, no effect of ovine GH or cortisol was seen in plasma osmolality or gill Na, K-ATPase activity when the hormones were given during the course of acclimation to dilute seawater. When catfish in fresh water were hypophysectomized, plasma osmolality was significantly decreased by 24 h, reaching a minimum level after 2 days. When they were transferred to dilute seawater, the plasma osmolality of the sham-operated fish was consistently higher than that of environmental water, whereas the osmolality of the hypophysectomized fish was equivalent to the environmental salinity. Ovine PRL restored the plasma osmolality of the hypophysectomized fish in fresh water to the level of sham-operated fish. Cortisol was also effective, but the effect was less pronounced than the effect of PRL. Injection of PRL in combination with cortisol resulted in a marked additive increase in plasma osmolality to a level even above that of the sham-operated fish. Ovine GH was without effect. These treatments in hypophysectomized fish transferred to dilute seawater produced essentially the same results as those in fish in fresh water. Plasma osmolality was also increased after PRL treatment of the intact fish in fresh water. There was a synergistic effect between PRL and cortisol in hypophysectomized fish in dilute seawater as well as in intact fish in fresh water. PRL did not stimulate cortisol secretion either in hypophysectomized fish or in intact fish. In the stenohaline catfish, both PRL and cortisol seem to be involved importantly in ion uptake from the environment not only in fresh water but also in brackish water.

Animals↗