Molecular Water Pump Research Articles

Molecular water pumps and the aetiology of Canavan disease: a case of the sorcerer's apprentice.

Molecular water pumps and the aetiology of Canavan disease: a case of the sorcerer's apprentice.

The existence of molecular water pumps in the nervous system: a review of the evidence

Recently, the presence of both influx and efflux molecular water pumps (MWPs) in vertebrate cells has been reported. These appear to use a common mechanism; the intercompartmental cotransport of water uphill against a gradient as a hydrophylic osmolyte is transported down its own gradient, in a regulated fashion, by a membrane spanning cotransporter protein. In each case, the dwell time of the transported osmolyte is short in that it is metabolically converted and its products either eliminated or recycled, thereby maintaining the required high intercompartmental gradient. An influx water pump osmolyte has been identified as a sodium-glucose complex, and an efflux water pump osmolyte as N-acetylhistidine. These osmolytes may also be archetypal representatives of many other osmolytes with similar functions in a variety of cells. When recycled, the osmolyte metabolites appear to be dewatered during high affinity binding that is associated with their active transport back across the membrane prior to intracellular resynthesis of the osmolyte. Since these cyclical systems result in the pumping of water, they also appear to create a previously unrecognized motive force which results in the establishment of unidirectional transcellular water flows between apical and basolateral cell membranes. As neurons represent highly specialized forms of animal cells, and cells which are also extremely sensitive to changes in osmotic pressure, the presence of these water pumps in the CNS could be significant. There would be connotations with regard to how neurons regulate water balance and transaxonal flow as well as to how these factors affect the integrated function of the nervous system. In this article, evidence of the presence of MWPs in the nervous system, and how they might relate to aspects of both normal and abnormal brain function is reviewed.

Expression of aspartoacylase activity in cultured rat macroglial cells is limited to oligodendrocytes.

N-acetyl-L-aspartate (NAA) is an important osmolyte in the vertebrate brain and eye, and its cyclical metabolism is accomplished in two separate compartments. In the brain, NAA is synthesized primarily in neurons, and after its regulated release, NAA is hydrolyzed by aspartoacylase, which is present in a glial-associated compartment. However, the precise nature of this hydrolytic compartment has remained obscure. It has been proposed that one role of aspartoacylase in the central nervous system (CNS) is as part of a molecular water pump (MWP) that uses the NAA intercompartmental cycle to remove nerve cell metabolic water against a water gradient and that oligodendrocytes comprise the second compartment in this metabolic sequence. The absence of aspartoacylase activity in Canavan disease (CD), a rare early onset genetic spongiform leukodystrophy, is associated with CNS edema, intramyelinic swelling and a progressive loss of oligdendrocytes. In order to evaluate the MWP hypothesis and its possible relationship to the etiology of CD further, both oligodendrocytes and astrocytes obtained from neonatal rat brain were grown in culture and tested for the presence of aspartoacylase activity. The results of this study show for the first time that aspartoacylase activity is expressed only in oligodendrocytes. The meaning of this observation in understanding the function of the NAA metabolic cycle is discussed.

Function of the N-acetyl-L-histidine system in the vertebrate eye. Evidence in support of a role as a molecular water pump.

N-acetyl-L-histidine (NAH) is a major constituent of poikilotherm brain, eye, heart, and muscle, but for which there is no known function. NAH is characterized by high tissue concentrations, a high tissue/extracellular fluid (ECF) gradient, and by a continuous selective and regulated efflux into ECF. In the eye, there is a complete compartmentalization of the synthetic and hydrolytic enzymes, with synthesis of NAH from AcCoA and L-histidine (His) occurring in the lens, and its hydrolysis to acetate and His restricted to surrounding ocular fluids. Using 14C-isotopes, the cycling of NAH between lens and ocular fluids in a simple support medium consisting of NaCl (0.9%), Ca2+ (4 mEq/L) and D-glucose (5 mM) at pH 7.4 has previously been observed. In the present study, using the isolated lens of the goldfish eye, each of the components of that support medium has been individually varied in order to determine its effect on NAH release down its intercompartmental gradient. As a result of these and related studies, it is suggested that NAH may function as a metabolically recyclable gradient-driven molecular water pump. It is proposed that water influx or generation of metabolic water serves as the trigger mechanism to open a Ca-dependent gate for the release of NAH down its gradient, along with its associated water. Preliminary analyses suggest that in addition to its potential for multiple daily cycles, a strongly ionized hydrophilic molecule, such as NAH, may include a large power function as a result of its attraction to water, and it has been calculated that an aqua complex of each NAH molecule may have 33 dipole-dipole-associated water molecules as it passes into ECF. It is this unique combination of a capacity for multiple cycles per day, coupled with a large power function, that may allow for such an intracellular osmolyte to be present in relatively low concentration in comparison to total cellular osmolality, and yet to perform a large and important task with little expenditure of energy. With each NAH molecule recycled up to 10 times/d, and a power factor of 33, there could be 330 mmol of water transported/mmol of NAH each day. With typical NAH concentrations in brains of poikilothermic vertebrates of 5-10 mmol/kg, there is the potential for up to 3.3 mol (60 mL) of water to be removed each day/kg of brain, a value that represents about 8% of total brain water content. Dewatering of the released osmolyte would occur in two additional steps, consisting of its hydrolysis and the subsequent active uptake of its metabolites. It is also suggested that NAH is the archetype of several metabolically and structurally related cellular osmolytes found in both poikilotherms and homeotherms, for which there is similarly no known function, and these may form a family of cycling hydrophilic osmolytes that serve as molecular water pumps in a variety of tissues. These include the basic His containing derivatives: NAH, carnosine, anserine, ophidine, and homocarnosine, and the acidic aspartate derivatives: N-acetyl-L-aspartate (NAA) and N-acetyl-L-aspartylglutamate (NAAG). In each of these cases, the high intracellular/extracellular osmolyte gradient appears to be maintained by combining a hydrophilic protein amino acid with a nonprotein moiety to block its use in other intracellular metabolic pathways, and by blocking catabolism of the derivative by maintaining its hydrolytic enzyme in an extracytosolic membrane or extracellular compartment. Unlike other known water-regulating mechanisms, the proposed cellular system is unique in that as a water pump, it can function as a water regulator independently of extracellular solute composition or osmolality. Finally, based on the hypothesis developed, the NAH system would represent the first cellular water pump to be identified.

The human Na+-glucose cotransporter is a molecular water pump.

1. The human Na+-glucose cotransporter (hSGLT1) was expressed in Xenopus laevis oocytes. The transport activity, given by the Na+ current, was monitored as a clamp current and the concomitant flux of water followed optically as the change in oocyte volume. 2. When glucose was added to the bathing solution there was an abrupt increase in clamp current and an immediate swelling of the oocyte. The transmembrane transport of two Na+ ions and one sugar molecule was coupled, within the protein itself, to the influx of 210 water molecules. 3. This stoichiometry was constant and independent of the external parameters: Na+ concentrations, sugar concentrations, transmembrane voltages, temperature and osmotic gradients. 4. The cotransport of water occurred in the presence of adverse osmotic gradients. In accordance with the Gibbs equation, energy was transferred within the protein from the downhill fluxes of Na+ and sugar to the uphill transport of water, indicative of secondary active transport of water. 5. Unstirred layer effects were ruled out on the basis of experiments on oocytes treated with gramicidin or other ionophores. Na+ currents maintained by ionophores did not lead to any initial water movements. 6. The finding of a molecular water pump allows for new models of cellular water transport which include coupling between ion and water fluxes at the protein level; the hSGLT1 could account for almost half the daily reuptake of water from the small intestine.

A molecular water pump in quartz dislocations

RECENTLY Bakker and Janssen1 have reported experiments suggesting that water leaks out of bubbles in quartz preferentially along dislocations against thermodynamic gradients in fugacity, chemical potential and pressure. This effect indicates that the density of gas-rich fluid inclusions cannot be calculated from metamorphic pressure-temperature conditions and, conversely, that the density of the fluid cannot be used to infer metamorphic pressure-temperature conditions. Using computer modelling techniques, I show here that water not only diffuses easily along dislocations in quartz, but can also be 'pumped' along them. The simulations indicate that in dislocation cores, water molecules dissociate and the protons and hydroxyl groups become strongly bound to kinks. A shear stress can do work on kinks to move them along a dislocation, dragging with them these water-bearing species and sweeping other, undissociated water molecules before them. © 1992 Nature Publishing Group.