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At least 19 recordsLinked to original sources
PubMed1978

Kinetics of the co-transport of phenylalanine and sodium ions in the guinea-pig small intestine. II. Sodium fluxes and flux ratios.

The validity of a general non-compulsory model for the description of the co-transport of sodium and phenylalanine in the small intestinal mucosa of the guinea-pig has been examined by measuring the influx of sodium and the flux ratios. The simultaneous influxes of sodium and phenylalanine have been determined by incubating intestinal tissues at a fixed sodium and variable phenylalanine concentrations. The experiment was repeated at a number of sodium concentrations, and a straight-line relationship between the fluxes was always obtained. The slope of this line, the flux ratio, is dependent on the sodium concentration; the function is biphasic inasmuch as it rises with sodium concentration at low external sodium, but decreases when the sodium concentration is raised above a level of about 70 mM. This response is compatible with the model under examination. No saturable component of sodium influx could be detected in the absence of phenylalanine, but the values of this influx corresponded with those predicted from the flux ratio experiment. In the presence of phenylalanine, sodium influx could be resolved into saturable and non-saturable components, and the Kt for sodium influx via the saturable mechanism agreed with that predicted from the constants derived from measurements of phenylalanine influx. The maximal velocity for sodium influx was similar to the maximal velocity for phenylalaline influx. Equations were derived to examine the behaviour of sodium influx as a function of the external phenylalanine concentration, and a Kt for phenylalanine was deduced which agrees closely with that obtained by studying phenylalanine fluxes directly. These results provide evidence in favour the applicability, in this species, of a model involving no compulsory pathway for the formation of a ternary complex between the carrier, a sodium ion and the phenylalanine molecule. Examination of the literature suggests that in other species different models may describe more accurately this co-transport mechanism.

Animals↗
PubMed1975

Chloride flux in bilayer membranes: the electrically silent chloride flux in semispherical bilayers.

High resistance semispherical bilayer membranes of areas as large as 0.3 cm-2 were formed from a decane solution of synthetic diphytanoylphosphatidylcholine. These bilayers had a specific resistance of about 10-9 omega cm-2 and a specific capacitance of 0.38 mu F cm- minus 2 at 20 degrees in 0.1 M KCL. Under these conditions, chloride permeability was 6.8 times 10- minus 8 cm/sec. This electrically silen 36-Cl flux was found to be about 10-3-fold larger than the chloride current calculated from the electrical parameters of the system. The chloride flux in the bilayer was independent of the applied electrical field and was unaltered by addition of reducing agents to the ambient aqueous solutions. It was, however, substantially reduced when NO3 minus was substituted for Cl minus on the side of the bilayer initially free of 36-Cl, or if I minus was added to the aquesous phases in the concentration range of 0.001-0.1 M. These results strongly suggested that the electrically silent flux of 36-Cl is primarily a carrier mediated diffusion process in which phosphatidylcholine acts as the carrier species.

Binding Sites↗
PubMed1979

Sodium and calcium fluxes in a clonal nerve cell line.

1. 22Na+ and 45Ca2+ fluxes were studied in the clonal nerve cell line PC12. Three distinct types of ion channels were found: (a) voltage-dependent Na+ channels, (b) voltage-dependent Ca2+ channels, and (c) acetylcholine-activated channels permeable to both ions. 2. 22Na+ uptake through voltage-dependent Na+ channels is induced by veratridine and scorpion venom, and is inhibited 50% by 5 X 10(-7) M-tetrodotoxin and greater than 98% by 5 X 10(-6) M-tetrodotoxin. 3. 45Ca2+ uptake through voltage-dependent Ca2+ channels is induced by depolarizing the cells in 50 mM-KCl. This flux is not dependent on the presence of Na+ in the medium and is insensitive to 5 X 10(-6) M-tetrodotoxin. However, 1 mM-Mn2+ causes a 95% inhibition of K+-induced 45Ca2+ uptake. 4. Veratridine and scorpion venom also induce voltage-dependent 45Ca2+ uptake which can be blocked by 1mM-Mn2+. In contrast to KCl-induced 45Ca2+ uptake, this flux is completely blocked by 5 X 10(-6) M-tetrodotoxin and is abolished by removal of Na+ from the medium. Thus the depolarizing stimulus for Ca2+ uptake in this case is Na+ from the medium. Thus the depolarizing stimulus for Ca2+ uptake in this case is Na+ influx through voltage-dependent Na+ channels. 5. Carbamylcholine induces both 22Na+ and 45Ca2+ fluxes which are blocked by nicotinic cholinergic antagonists with the exception of alpha-bungarotoxin. The 22Na+ flux occurs exclusively via acetylcholine receptor channels, as evidenced by the lack of effect of 5 X 10(-6) M-tetrodotoxin. In the presence of Na+, almost all of the 45Ca2+ uptake can be blocked by 1 mM-Mn2+ and thus occurs via voltage-dependent Ca2+ channels which are activated by the depolarizing Na+ influx. 6--8% of the total 45Ca2+ flux, however, is insensitive to 1 mM-Mn2+, suggesting that this portion of the uptake occurs via the acetylcholine receptor channels. In Na+-free medium, the Mn2+-resistant 45Ca2+ component increases to 40% of the total uptake, apparently due to lack of competition from Na+ for the acetylcholine receptor channels. This receptor-linked flux still causes sufficient depolarization to induce the additional 60% of the Ca2+ flux through voltage-dependent, Mn2+ sensitive Ca2+ channels. 6. Mn2+ inhibits Ca2+ flux through voltage-dependent Ca2+ channels by competing for entry through these channels. 50 mM-KCl induces 54Mn2+ fluxes in PC12 cells that are comparable in magnitude to 45Ca2+ fluxes. 7. In normal saline 45Ca2+ efflux from PC12 cells is several times more rapid than in Na+-free medium, indicating the presence of a Ca2+-Na+ exchange mechanism.

Animals↗
PubMed1976

Relations between the electrical potential, pH gradient, proton flux and phosphorylation in the photosynthetic membrane.

The transmembrane electrical potential (deltaphi), the proton flux (H+), the rate of electron transport (e), the pH gradient (deltapH) and the rate of phosphorylation (ATP) were measured in chloroplasts of spinach. Photosynthesis was excited periodically with flashes of variable frequencies and intensities. A new method is described for determining the rate of electron transport and proton flux. Under conditions where the rate of electron transport and proton flux are not pH controlled the following correlations were found in the range 50 mV less than or equal to deltaphi less than or equal to 125 mV and 1.8 less than or equal to deltapH less than or equal to 2.7: (1) The pH gradient, deltapH, increases with H+ independently of Phout between 7-9. (2) The rate of phosphorylation, ATP, depends exponentially on deltapH (at constant deltaphi) and is independent of pHout between 7-9. (3) The rate of phosphorylation, ATP, depends also on deltaphi (at constant deltapH and at constant proton flux H+). (4) The proton flux via the ATPase pathway, Hp+, depends non-linearly on the ratio of the proton concentrations: Hp+ approximately (Hin+/Hout+)b, (b=2.3--2.6). The proton flux via the basal pathway, Hb+, depends linearly on the ratio of the proton concentrations: Hb+ approximately (Hin/Hout). (5) The ratio deltaH+/ATP (e/ATP, i.e. the ratio of the total proton flux, Hp+ + Hb+, and the rate of ATP formation, ATP, depends strongly on deltaphi and on deltapH. The ratio is deltaH+/ATP approximately 3 (e/ATP approximately 1.5) at deltapH 2.7 and deltaphi = 125 mV. (6) It is supposed that the reason for the dependence of deltaH+/ATP on deltaphi anddeltapH is the different functional dependence of the basal proton flux Hb+ and the phosphorylating proton flux Hp+ on deltapH and deltaphi. The calculation of deltaH+/ATP on the basis of this assumption is in fair agreement with the experimental values. Also the "threshold" effects can be explained in this way. (7) The ratio of deltaHp+/ATP, i.e. the ratio of the phosphorylating proton flux Hp+ and ATP, is deltaHp+/ATP APPROXIMATELY 2.4.

Adenosine Triphosphate↗
PubMed1979

Simultaneous minute by minute determination of unidirectional and net water fluxes in frog urinary bladder. A reexamination of the two barriers in series hypothesis.

Unidirectional and net water fluxes were simultaneously estimated in frog urinary bladder. The minute by minute tritiated water (3HOH) transepithelial flux and the net volume of fluid traversing the tissue were employed. It was observed that: (1) the time course of the increase in the 3HOH flux induced by antidiuretic hormone had a very similar pattern to that reported for the increase in the net movement. (2) Unstirred layers strongly affected the magnitude of the antidiuretic hormone-induced increase in 3HOH fluxes while the time course of the response was almost non-affected. In non-stimulated bladders 3HOH fluxes were poorly modified by medium stirring. New steady-state conditions for 3HOH fluxes were established 1 min after stirring rate modifications. (3) The simultaneously determined net water flux was not affected by a modification in the unstirred layers, indicating that the variations in the measured net water fluxes are a good estimation of the changes in the mucosal border permeability. (4) The presence of an osmotic gradient during hormonal challenge (implying net water fluxes, cell swelling and dilation of the intracellular spaces) did not modify the time course of 3HOH movements. These results suggest that the time course of the increase in water permeability is an intrinsic characteristic of the experimental system that could result from the addition of permeability units that increase in number during the development of the hormonal action.

Animals↗
PubMed1979

86Rb+ fluxes in Chinese hamster ovary cells as a function of membrane cholesterol content.

Steady-state fluxes of 86Rb+ (as a tracer for K+) were measured in Chinese hamster ovary cells (CHO-K1) and a mutant (CR1) defective in the regulation of cholesterol biosynthesis; the membrane cholesterol content of this mutant was varied by growing it on a range of cholesterol supplements to lipid-free medium (Sinensky, M. (1978) Proc. Natl. Acad. Sci. U.S. 75, 1247--1249). Analogous to previous findings in ascites tumor cells, 86Rb+ influx in the parent strain was differentiated into a ouabain-inhibitable 'pump' flux, furosemide-sensitive, chloride-dependent exchange diffusion, and a residual 'leak' flux. On the basis of this flux characterization, 86Rb+ pump and leak fluxes were measured in the mutant as a function of membrane cholesterol content. Pump and leak fluxes, when expressed per ml cell water, were independent of the cholesterol content of the mutant. Moreover, 86Rb+ fluxes in the mutant were equal to those in the parent strain. Our data imply that the flux behavior of K+ in the steady state is independent of the ordering of membrane lipid acyl chains.

Animals↗
PubMed1975

Chloride flux in bilayer membranes: chloride permeability in aqueous dispersions of single-walled, bilayer vesicles.

Aqueous dispersions of phosphatidylcholine vesicles were utilized to determine bilayer permeability to 36-Cl as a function of pH and temperature. These dispersions were comprised of single-walled vesicles, homogeneous in size, prepared by sonication of purified egg phosphatidylcholine under argon followed by fractionation on a molecular sieve. Permeability constants calculated from the inward flux of 36-Cl and the geometric parameters of these vesicles proved to be dependent on both pH and temperature. Analysis of these dependences leads to the conclusion that 36-Cl permeation in the presence of KCl is due principally to a carrier mediated exchange process involving a phospholipid-HCL complex. Net permeation by H-36-Cl may make a small contribution to the 36-Cl flux, however, studies carried out at very low chloride concentrations show that this flux is much smaller than the exchange flux. Thus chloride permeability for the exchange process is 1.5 times 10- minus 11 cmsec- minus 1 while the corresponding coefficient for the net flux of H-36-Cl is 1.0 times 10- minus 12 cm sec- minus 1 at pH 7. The activation energy for the 36-Cl exchange flux was found to be 19 plus or minus 2 kcal/mol. This value is similar to that obtained for the transbilayer "flip-flop" of phosphatidylcholine molecules in a similar system (Kornberg and McConnell, 1971). This correspondence together with the fact that the experimentally determined flux of 36-Cl agrees well with that calculated from the "flip-flop" parameters, strongly suggests that the flux of 36-Cl and "flip-flop" of phosphatidylcholine may be the same process.

Binding Sites↗
PubMed1977

Effect of perfusion rate on the fluxes of water, sodium, chloride and urea across the proximal convoluted tubule.

Studies were undertaken to examine the mechanism whereby changes in intraluminal flow rates after reabsorption in the isolated perfused proximal tubule of the rabbit. All protocols employed the technique of in vitro perfusion of isolated segments of the proximal convoluted tubule. Stepwise elimination of d-glucose and l-alanine from an artifical perfusate stimulating ultrafiltrate decreased the unidirectional flux of sodium, transtubular potential difference, and net water absorption. Using isosmolal ultrafiltrate as the perfusate, net fluid reabsorption and the unidirectional lumen-to-bath flux of sodium and chloride decreased with a decrease in flow rate below 11 nl/min, but neither net fluid reabsorption nor the unidirectional fluxes of sodium and chloride increased further as the perfusion rate was increased above 11 nl/min. The unidirectional flux of 14C-urea was not affected by changes of perfusion rate from 1.6 to 44 nl/min. The dependence of net fluid reabsorption and unidirectional fluxes of sodium and chloride on flow rate per se, and not on intraluminal hydrostatic pressure or geometry, was established by demonstrating their decrease despite a rise in intraluminal pressure and inside diameter produced by counterpressure at the collecting end of the tubule, while flow was decreased. Ouabain decreased net fluid reabsorption to near zero at all flow rates, but ouabain had no effect on the flow-dependency of unidirectional sodium anf sodium was eliminated with a decrease in bicarbonate concentration and removal of d-glucose and l-alanine from the perfusate. Thus, the present studies demonstrate that net water and unidirectional sodium and chloride fluxes are flow-dependent. At flow rates somewhere below 11 nl/min, unidirectional fluxes decreased with decreasing perfusion rates; however, at perfusion rates greater than 11 nl/min, there was no further effect of perfusion rate on either net water absorption or the unidirectional fluxes of sodium or chloride. These effects may be partly mediated through the flow-dependent changes in the intraluminal concentration of bicarbonate, d-glucose, and 1-alanine.

Adsorption↗
PubMed1977

Tracer Na fluxes in Necturus proximal tubule.

Steady-state bidirectional sodium fluxes were measured across Necturus proximal tubules. New methods for capillary perfusion and collection of venous effluent enabled flux determination to be made from the appearance of luminal tracer in the capillaries. Fluxes and permeability were measured in the absence of net fluid reabsorption. The sodium permeability measured in the plasma-to-lumen direction was 3 X 10(-6) cm/s. The flux ratio (lumen-to-plasma/plasma-to-lumen) was about twice the passive value calculated from the measured concentrations and potentials. Estimates for the permeability and flux across the shunt pathway were obtained from nonsteady-state flux determinations. The shunt pathway appeared to be the most significant route for passive sodium movement from plasma-to-lumen. Nonsteady-state tracer measurements also enabled an estimate to be made of the lumenal cell membrane permeability and unidirectional sodium flux. Two-thirds of the lumen-to-plasma flux was calculated to traverse the cellular path and the remainder through the shunt. Approximately one-third of the intracellular sodium was found to exchange rapidly with tracer.

Animals↗
PubMed2025

Suppression of OTUD4 protects against myocardial ischemia-reperfusion injury by increasing autophagic flux and inhibiting apoptosis in cardiomyocytes.

Dysregulated autophagic flux plays a critical role in myocardial ischemia-reperfusion injury (MIRI), complicating cardiac reperfusion therapy. In this study, we identified OTUD4 as a potential regulator of autophagic flux in MIRI using CRISPR/Cas9 sgRNA sequencing. However, the underlying mechanism is poorly understood. The purpose of this study is to investigate the effects of OTUD4 on autophagic flux in OGD-R treated AC16 cells (IRI model in vitro) and LAD artery ligation induced myocardial ischemia-reperfusion mice (MIRI model in vivo). In the in vitro IRI cell model, OTUD4 knockdown significantly reversed impaired autophagic flux, increased mitochondrial membrane potential, and decreased LDH activity, ROS production, autophagy and apoptosis. Overexpression of OTUD4 showed the opposite result. In the in vivo MIRI model, OTUD4 knockdown also significantly decreased infarct area, improved cardiac structure and function, reduced serum BNP and LDH levels, attenuated cardiac tissue injury/fibrosis/myocardial hypertrophy, and ultimately exerted myocardial protective effects against ischemia-reperfusion injury. Importantly, OTUD4 knockdown inhibited autophagosome-associated markers (LC3II/LC3I, Beclin1, ATG9), autophagy substrate p62, increased lysosomal activity marker LAMP2, and activated the autophagy pathway (AKT/mTOR), thereby promoting the recovery of impaired autophagic flux in the MIRI model. Moreover, OTUD4 showed strong interaction with UBAC1, and OTUD4 deficiency decreases UBAC1 protein expression by impairing its deubiquitination, thereby regulating autophagy. In short, blocking OTUD4 restored damaged autophagic flux in I/R induced myocardial injury both in vivo and in vitro, inhibited myocardial cell apoptosis, and greatly improved cardiac function in ischemia-reperfusion mice. KEY MESSAGES: OTUD4 was identified as a key negative regulator of autophagy flux in myocardial ischemia-reperfusion injury (MIRI) via genome-wide CRISPR/Cas9 screening. OTUD4 knockdown exerts cardioprotective effects by reducing apoptosis and ROS generation and improving heart function in both in vitro and in vivo models. The interaction between OTUD4 and UBAC1 was confirmed, and OTUD4 maintains UBAC1 stability through deubiquitination, providing new insights into the ubiquitination regulatory mechanism in myocardial injury. Targeting OTUD4 has therapeutic potential for MIRI, as OTUD4 knockdown alleviated MIRI in both in vitro and in vivo models, suggesting the possibility of developing OTUD4 inhibitors for cardiac reperfusion treatment.

Animals↗
PubMed1978

Use of flux ratio measurements for the determination of the order of addition of substrates and products in enzyme reactions.

1. Methods of determining the order of addition of substrates and dissociation of products by using flux ratios are investigated. Where an enzyme obeys hyperbolic steady-state velocity kinetics it is concluded that it may be particularly useful to compare the measured flux ratios with those calculated from the steady-state velocity parameters. 2. An expression is derived relating the relative contribution of the two pathways in a branched pathway to the flux ratios. 3. The relationship of equilibrium-reaction-rate measurements [Boyer & Silverstein (1963) Acta Chem. Scand. 17, Suppl. 1, S195] to the flux ratios is considered. Equilibrium-reaction rates are shown to be affected both by the addition of substrates and dissociation of products. Methods of analysing the data to distinguish between these events are discussed. 4. Methods of measurement of flux ratios are described, and it is concluded that the non-equilibrium steady-state method is preferable to measurements at chemical equilibrium. 5. The relative significance of flux ratio measurements and steady-state velocity inhibition data is discussed. It is concluded that flux ratios, when taken in conjunction with the inhibition data, provide the least ambiguous information about mechanism.

Enzymes↗
PubMed1975

Relation between insulin and glucose flux rates in the dog.

A constant infusion isotope dilution procedure for measurement of plasma insulin flux rates was performed in acutely eviscerated dogs receiving an infusion of nonlabeled insulin; the calculated rate of insulin flux corresponded closely to that of the administered rate, thus validating the isotope dilution procedure. Flux rates of insulin and glucose were measured simultaneously at different steady levels in intact dogs. There was a linear relationship between plasma insulin concentration and insulin flux rate. In each animal there was also a linear relationship between glucose flux rate and insulin concentration, and between rates of glucose flux and insulin flux, but there was considerable inter-animal variation in the slope of the regression lines.

Animals↗
PubMed1977

Effect of heat stable and heat labile Escherichia coli enterotoxins, cholera toxin and theophylline on unidirectional sodium and chloride fluxes in the proximal and distal jejunum of weanling swine.

Acute, isolated loops of proximal and distal jejunum of weanling swine were exposed to either heat stable porcine Escherichia coli enterotoxin, heat labile porcine Escherichia coli enterotoxin, cholera toxin or theophylline. Unidirectional sodium fluxes in response to heat stable in the proximal jejunum were dependent on the length of time that the intestinal mucosae was exposed to the enterotoxin. Net water, sodium and chloride and unidirectional sodium and chloride flux measurements in the proximal jejunum in response to each agent uniformly indicated that net secretion of fluid and electrolytes was the result of increased unidirectional sodium secretion or blood-to-lumen flux and decreased unidirectional chloride absorption or lumen-to-blood flux. In addition heat stable cholera toxin and theophylline but not heat labile decreased unidirectional chloride secretion a small but significant amount in the proximal jejunum. Sodium and chloride flux measurements in the distal jejunum demonstrated that all four secretory agents could stimulate net secretion of water, sodium and chloride in that region. The response to these secretory agents as measured by sodium and chloride unidirectional flux rates was not similar to changes observed in the proximal jejunum. In the distal small intestine, whereas heat labile cholera toxin and theophylline induced similar qualitative changes in unidirectional sodium and chloride fluxes, that induced by heat stable differed.

Animals↗
PubMed1977

Quantitative analysis of the change of metabolite fluxes along the pentose phosphate and glycolytic pathways in Tetrahymena in response to carbohydrates.

A metabolic scheme of glycolysis and the pentose phosphate pathway has been constructed, assuming that the reactions occur in a single compartment. From this scheme, equations are written for a system in metabolic and isotopic steady state. These allow computation of the specific activity of every carbon atom of all the intermediates of the glycolytic and pentose phosphate pathways and consequently of the flux of carbon along each step of these pathways. A sufficiently large number of well distributed measurements of incorporation of radioactive label from different positions of several substrates into intermediates or products must be made to determine all the fluxes. This is done by choosing a set of metabolic fluxes, calculating incorporation with the aid of a computer, and then manipulating the flux rates until the computed incorporations match the data. The model is used in this paper to analyze the metabolism of the protozoan Tetrahymena pyriformis. The metabolic scheme of the model is consistent with all available information on the enzyme complement of this ciliate. Cells grown to transition phase in proteose/peptone medium were inoculated into a mixture of glucose (6 mM), fructose (6 mM), ribose (3 mM), and glycerol (3 mM) and incubated for 1 h. In each of these experiments, one of the following labeled substrates was present: [1-, 2-, 6-, or U-14C]glucose; [1- or U-14C]fructose; [1- or U-14C]ribose; [1(3)-or 2-14C]glycerol. The incorporation of label from these substrates into CO2, lipid, glycogen, and RNA was measured. In contrast to earlier studies on the metabolism of 2- and 3-carbon substrates by Tetrahymena, the rate of incorporation of label from some substrates into some products (e.g. from [1-14C]glucose into CO2) changed during the incubation. To treat these time-dependent data within the framework of the steady state model, the 1-h incubation was divided into three 20-min intervals; within each of these, the rates of incorporation were approximately constant, as required for a steady state system. Measurements of the pool sizes of glucose-6-P and fructose-6-P showed that only slow changes in pool sizes occurred after the first 5 min of incubation and indicated that the system was effectively in a metabolic and isotopic steady state throughout most of the incubation. The finding that a low concentration of cycloheximide prevented the acceleration of 14CO2 production from labeled glucose suggests a role for protein synthesis in the slow adaptation to carbohydrate addition and supports the quasi-steady state treatment of this system. The expected incorporation into each product was computed for trial sets of 1, independent flux rates. A set of flux values was found which yielded a good fit to the 29 measurements made for each interval. These flux values therefore constitute a quantitative description of temporal changes in carbon flow along the glycolytic and pentose phosphate pathways during the 1st h of adaptation to the carbohydrate mixture...

Adenosine Triphosphate↗
PubMed2026

13C Stable Isotope Tracing-Based MFA Reveals the Contribution of Glucose to Glycolytic and TCA Fluxes and Its Application in Depression Research.

Metabolomics is widely applied to dissect metabolic pathways and their correlations with biological phenotypes. Unlike genomics and proteomics, metabolites exhibit substantial heterogeneity in chemical structure, physicochemical properties, and biological origin. Accordingly, pathway enrichment and annotation relying merely on alterations in metabolite abundance are prone to incomplete coverage, ionization bias, and ambiguous annotation, which inevitably impair the accuracy of pathway interpretation. Metabolic flux analysis (MFA) coupled with stable isotope-resolved metabolomics (SIRM) offers a powerful quantitative framework for tracing in vivo carbon flow and estimating reaction fluxes across key metabolic nodes. Glucose metabolism lies at the core of systemic energy homeostasis; however, most current investigations are confined to cell lines or in vitro systems, and a simple, easy-to-implement computational pipeline for in vivo glucose flux analysis in animal models is still lacking. Herein, we established an in vivo 13C-labeling-based MFA workflow to trace and resolve the systemic metabolic fate of glucose in rats. The pipeline covers tracer administration, sample preparation, LC-MS detection, isotopologue data acquisition and correction, construction of a glucose-metabolism-related metabolite database, MFA model establishment, and metabolic flux quantification. By infusing rats with [U-13C6]-glucose and [U-13C3]-sodium L-lactate, we precisely characterized the in vivo metabolic fates of circulating glucose and lactate and quantified their respective contributions to glycolytic flux and tricarboxylic acid (TCA) cycle flux. We further applied this workflow to profile energy metabolic reprogramming in depression. The results revealed a systemic shift toward aerobic glycolysis in rats exposed to chronic unpredictable mild stress (CUMS). Overall, the expanded application of this MFA strategy can provide mechanistic and quantitative insights into the regulation of metabolic pathways.

Animals↗
PubMed1977

Water fluxes in nerve fiber.

The hydrostatic (Lp) and osmotic (LPD) filtration coefficients and the efflux rates of tritiated water were measured in the giant axon of Loligo vulgaris. The Lp was 8 to 14 X 10(-8) cm/sec/cm H2O and the LPD was two orders of magnitude smaller (3 to 6 X 10(-10) cm/sec/cm H2O). In axons whose diameter was approximately 500 micron, the time (t1/2) required for a reduction in the axonal labeled water activity to one half its initial value was 38 to 48 sec. The rate limiting structure for solute flux was made ineffective by (1) storing the axon in isosmotoc KF at 0-2 degrees C for one month to one year or by (2) fixing the axon in 2-4% glutaraldehyde for 3 to 7 hr. The criteria of ineffectiveness of the rate limiting structure for solute flux were (1) a reduction of LPD to immeasurably low values, (2) the absence of electrical properties characteristic of plasmalemma, and (3) a marked increase in the rate of efflux of Na22. In such impaired axons the Lp and the t 1/2 of tritiated water efflux were unaffected. This independence of solute and solvent flux in conjunction with the finding that the hydraulic conductivity determined by bulk osmotic and hydrostatic pressure gradients is not equivalent (i.e., LPD/LP less than 1) indicate that the rate limiting structures for solute and solvent flux are in series. Solvent fluxes appear to be surface-limited, not bulk-limited. We have been unable to resolve whether the surface structure involved in limiting solvent flux is the sheath (Schwann layer and adhering connective tissue) and/or the cortical layer of the axoplasmic gel.

Animals↗
PubMed1976

Deviations from the flux ratio equation for chloride ions in ouabain- and acetazolamide-treated frog skin.

In order to establish whether or not chloride ions behave as freely moving particles in "passive", i.e. ouabain- and acetazolamide-treated, frog skin, tracer fluxes of 36Cl-have been measured while a voltage (generally +40 mV, serosal side positive) across the skin was applied. Ussing's flux ratio equation has been used as a criterion for this type of transport. One group of skin samples exhibited significant exchange diffusion phenomena. Most samples in a second group either behaved according to the flux ratio equation of showed significant and extreme exchange diffusion. From flux ratios obtained at two different voltages across various skin samples, showing extreme exchange diffusion, it appeared that the simple form of Kedem and Essig's law derived from irreversible thermodynamics, which is valid for homogeneous systems, does not apply to the type of exchange diffusion found. The system can, however, be described by a 1:1 exchange mechanism working in parallel with a diffusional pathway. The ratio exchange flux/observed efflux must then have a constant value (0.83) at the voltages appled, which implies that the exchange flux is voltage dependent. By comparison with iodide flux experiments as carried out by Ussing, it is shown that iodide exhibits the same type of exchange diffusion. A carrier, possibly responsible for the observed behaviour, is described.

Acetazolamide↗