Increase In Passive Permeability Research Articles

Paradoxical Increase of Permeability and Lipophilicity with the Increasing Topological Polar Surface Area within a Series of PRMT5 Inhibitors.

An imidazolone → triazolone replacement addressed the limited passive permeability of a series of protein arginine methyl transferase 5 (PRMT5) inhibitors. This increase in passive permeability was unexpected given the increase in the hydrogen bond acceptor (HBA) count and topological polar surface area (TPSA), two descriptors that are typically inversely correlated with permeability. Quantum mechanics (QM) calculations revealed that this unusual effect was due to an electronically driven disconnect between TPSA and 3D-PSA, which manifests in a reduction in overall HBA strength as indicated by the HBA moment descriptor from COSMO-RS (conductor-like screening model for real solvation). HBA moment was subsequently deployed as a design parameter leading to the discovery of inhibitors with not only improved passive permeability but also reduced P-glycoprotein (P-gp) transport. Our case study suggests that hidden polarity as quantified by TPSA-3DPSA can be rationally designed through QM calculations.

Deletion of the Imprinted Phlda2 Gene Increases Placental Passive Permeability in the Mouse.

Genomic imprinting, an epigenetic phenomenon that causes the expression of a small set of genes in a parent-of-origin-specific manner, is thought to have co-evolved with placentation. Many imprinted genes are expressed in the placenta, where they play diverse roles related to development and nutrient supply function. However, only a small number of imprinted genes have been functionally tested for a role in nutrient transfer capacity in relation to the structural characteristics of the exchange labyrinthine zone. Here, we examine the transfer capacity in a mouse model deficient for the maternally expressed Phlda2 gene, which results in placental overgrowth and a transient reduction in fetal growth. Using stereology, we show that the morphology of the labyrinthine zone in Phlda2−/+ mutants is normal at E16 and E19. In vivo placental transfer of radiolabeled solutes 14C-methyl-D-glucose and 14C-MeAIB remains unaffected at both gestational time points. However, placental passive permeability, as measured using two inert hydrophilic solutes (14C-mannitol; 14C-inulin), is significantly higher in mutants. Importantly, this increase in passive permeability is associated with fetal catch-up growth. Our findings uncover a key role played by the imprinted Phlda2 gene in modifying placental passive permeability that may be important for determining fetal growth.

Immune responses at brain barriers and implications for brain development and neurological function in later life

For a long time the brain has been considered an immune-privileged site due to a muted inflammatory response and the presence of protective brain barriers. It is now recognized that neuroinflammation may play an important role in almost all neurological disorders and that the brain barriers may be contributing through either normal immune signaling or disruption of their basic physiological mechanisms. The distinction between normal function and dysfunction at the barriers is difficult to dissect, partly due to a lack of understanding of normal barrier function and partly because of physiological changes that occur as part of normal development and ageing. Brain barriers consist of a number of interacting structural and physiological elements including tight junctions between adjacent barrier cells and an array of influx and efflux transporters. Despite these protective mechanisms, the capacity for immune-surveillance of the brain is maintained, and there is evidence of inflammatory signaling at the brain barriers that may be an important part of the body's response to damage or infection. This signaling system appears to change both with normal ageing, and during disease. Changes may affect diapedesis of immune cells and active molecular transfer, or cause rearrangement of the tight junctions and an increase in passive permeability across barrier interfaces. Here we review the many elements that contribute to brain barrier functions and how they respond to inflammation, particularly during development and aging. The implications of inflammation–induced barrier dysfunction for brain development and subsequent neurological function are also discussed.

Luminal hyperosmolarity decreases Na transport and impairs barrier function of sheep rumen epithelium

The effects of luminal hyperosmolarity on Na and Cl transport were studied in rumen epithelium of sheep. An increase of luminal osmotic pressure with mannitol (350 and 450 mosm/l) caused a significant increase of tissue conductance, G (T), which is linearly correlated with flux rates of (51)Cr-EDTA and indicates an increase of passive permeability. Studies with microelectrodes revealed, that an increase of the osmotic pressure caused a significant increase of the conductance of the shunt pathway from 1.23 +/- 0.10 (control) to 1.92 +/- 0.14 mS cm(-2) (450 mosm/l) without a change of fractional resistance. Hyperosmolarity significantly increased J (sm) and reduced J (net) Na. The effect of hyperosmolarity on J (ms) Na is explained by two independent and opposed effects: increase of passive permeability and inhibition of the Na(+)/H(+) exchanger. Hypertonic buffer solution induced a decrease of the intracellular pH (pH(i)) of isolated ruminal cells, which is consistent with an inhibition of Na(+)/H(+) exchange, probably isoform NHE-3, because NHE-3-mRNA was detectable in rumen epithelium. These data are in contrast to previous reports and reveal a disturbed Na transport and an impaired barrier function of the rumen epithelium, which predisposes translocation of rumen endotoxins and penetration of bacteria.

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We have studied the effect of polarity, current density, current duration, crosslinking density, swelling ratio, and permeation enhancers on the transdermal flux of ketoprofen from acrylamide hydrogel. Hydrogel was prepared by free radical crosslinking polymerization of acrylamide. Drug loading was made just before transport experiment by soaking the hydrogel in solution containing drug. In vitro flux study using hairless mouse skin was performed at using side-by-side diffusion cell, and the drug was analysed using HPLC/UV system. The result showed that, compared to passive flux, the total amount of drug transported increased about 18 folds by the application of cathodal current. Anodal delivery with same current density also increased the total amount of drug transported about 13 folds. It seemed that the increase in flux was due to the electrorepulsion and the increase in passive permeability of the skin by the current application. Flux increased as current density, the duration of current application and loading amount (swelling duration) increased. As the cross linking density of the hydrogel increased, flux clearly decreased. The effect of hydrophilic enhancers (urea, N-methyl pyrrolidone, Tween 20) and some hydrophobic enhancers (propylene glycol monolaurate and isopropyl myristate) was minimal. However, about 3 folds increase in flux was observed when 5% oleic acid was used. Overall, these results provide some useful information on the design of an optimized iontophoretic delivery system of ketoprofen.

Plasma membrane H-ATPase activity is involved in adaptation of tomato calli to NaCl.

A tomato (Lycopersicon esculentum Mill. cv. Pera) callus culture tolerant to NaCl was obtained by successive subcultures of NaCl-sensitive calli in medium supplemented with 50 mM NaCl. NaCl-tolerant calli grew better than NaCl-sensitive calli in media supplemented with 50 and 100 mM NaCl. Analysis of callus ion content showed a strong increase in Na+ and Cl- both in NaCl-tolerant and -sensitive calli grown in media containing NaCl for one subculture. Cells from NaCl-tolerant calli showed a higher H+ extrusion activity than those from NaCl-sensitive calli grown for one subculture in the presence of NaCl. The inhibition of H+ extrusion by NaCl-sensitive cells was correlated with an inhibition of microsomal vanadate-sensitive H+-ATPase (EC 3.6.1.35) and ATP-dependent H+ transport, while the stimulation of H+ extrusion by cells tolerant to 50 mM NaCl was correlated with an increase in plasma membrane ATP-dependent H+ transport. The increase of ATP-dependent H+ extrusion in plasma membranes isolated from 50 mM NaCl-tolerant calli was not a result of stimulation of a vanadate-sensitive ATP hydrolytic activity or an increase in passive permeability to H+. Relative to NaCl-sensitive calli, plasma membrane H+-ATPase from calli tolerant to 50 mM NaCl showed a lower Km for Mg2+-ATP. Our results indicate that tolerance of tomato calli to 50 mM NaCl increases the affinity of plasma membrane H+-ATPase for the substrate ATP and stimulates the H+-pumping activity of this enzyme without modifying its phosphohydrolytic activity.

Influence of the glycopeptidic moiety of mycobacterial glycopeptidolipids on their lateral organization in phospholipid monolayers

Glycopeptidolipids (GPLs) from the cell wall of opportunistic pathogenic mycobacteria are potential factors of pathogenicity which can interact with biological membranes. GPL suspensions uncouple oxidative phosphorylation of mitochondria and increase membrane permeability of liposomes. Heavily glycosylated GPLs are less active than lightly glycosylated ones. GPL–phospholipid interactions into preformed mixed films at the air–water interface were investigated in order to understand the permeabilization efficiency differences among GPLs. Polarization modulation infrared reflection absorption spectroscopy (PMIRRAS) was used to determine, in situ, the organization of GPL and of 1,2-di(perdeuteropalmitoyl)phosphatidylcholine (DPPC) molecules in mixed films. Compression isotherms of GPL alone or mixed with DPPC in various proportions showed that the less the GPL was glycosylated the higher its miscibility with DPPC. PMIRRAS studies indicated that low miscibility may result from large self-association of GPL molecules in β-sheet structures. Low glycosylated GPL molecules increased disorder of DPPC acyl chains. Based on these results, an explanatory model is proposed for membrane permeabilization. Increase of passive permeability may arise from disruption of phospholipid packing induced by GPL molecules. GPL segregation is proposed as the cause of low activity of GPL with high sugar content, by decreasing the number of GPL molecules interacting with phospholipids.

Two British families with variants of the 'cryohydrocytosis' form of hereditary stomatocytosis.

We describe two British families with similar, dominantly-inherited, temperature-related variants of hereditary stomatocytosis, consistent with the original description of 'cryohydrocytosis'. The cells show a 5-6-fold increase in passive permeability at 37 degrees C with abnormal intracellular Na and K levels at 15-20 and 60-65 mmol/(l cells) respectively. Marked temperature effects were evident: lysis of red cells on storage in the cold was blatant and when whole heparinized blood was stored at room temperature, K accumulated in the plasma, producing 'pseudohyperkalaemia'. Studies of the temperature dependence of passive permeability showed that the minimum in the passive permeability, which is seen in normal cells at 8-10 degrees C, was shifted up to 23 degrees C in these abnormal cells, such that the permeability at 0 degrees C exceeded that at 37 degrees C. The abnormal temperature dependence in these genetically abnormal red cells strongly resembles that seen in normal cells when suspended in media in which either Na or Cl has been replaced by an organic cation or anion: it could be said these cells had a genetic mutation that somehow rendered the cell resistant to the stabilizing action of NaCl at low temperatures.

Oxidative burst and electrolyte leakage induced by sulfhydryl blockers and by membrane permeabilizing reagents in different organs ofEgeria densa

Sulfhydryl blockers, such as N-ethylmaleimide, iodoacetate and heavy metals induce a transitory stimulation of O2 consumption and H2O2 production (oxidative burst) and a rapid release of electrolytes in leaves of various aquatic plants. The correlation between these two responses to N-ethylmaleimide or to Ag+ in separate organs and stages of leaf development was investigated inEgeria densa. Only adult leaves were able to respond to the sulfhydryl blockers with an oxidative burst, whereas this response was absent in immature growing leaves and in stem and root segments. In N-ethyl-maleimide- as well as in Ag+-treated adult leaves the oxidative burst was constantly associated with a relevant electrolyte leakage. These data are consistent with a model in which the SH reagent would first interact with a plasmalemma protein, leading to an increase in passive permeability to ions and to the activation of an oxidative enzyme of the type of the superoxide synthase described for granulocytes. In its turn, active-oxygen species produced by the activated oxidase might further damage the plasma membrane, increasing its passive permeability. Digitonin and nystatin, two reagents known to cause a permeabilization of lipid membranes, induced in adultE. densa leaves a transient increase in the rate of O2 consumption and H2O2 production and an electrolyte leakage very similar to those induced by sulfhydryl blockers. These effects, however, were not influenced by the flavin analogues diphenylene iodonium and quinacrine, and were partially inhibited by the presence of CN− and salicylhydroxamic acid, thus suggesting the involvement of a different oxidase in the oxidative burst elicited by these reagents.

A Method to Quantitatively Compare In Vivo the Effects of Gallstone Solvents on Intestinal Mucosal Function: A Controlled Study Comparing Mono-Octanoin with Methyl tert-Butyl Ether in the Rat

During contact dissolution of gallstones, solvents may escape from the gallbladder and damage the intestinal mucosa. In order to compare the extent of this potential injury, we developed a method to objectively quantify the effects of two commonly used cholesterol solvents, methyl tert-butyl ether and mono-octanoin, on mucosal transport function in the rat intestine. Two intestinal segments in each of 184 anesthetized rats were cannulated. Three milliliters of either solvent were instilled in one segment and left for varying periods of time, while saline was instilled in the other as control. The segments were then washed and perfused for 45 min with an isotonic solution containing [3H]polyethylene glycol 4000 (a nonabsorbable reference marker) and either [14C]alpha-aminoisobutyric acid (a marker for active absorption) or [14C]mannitol (a marker for passive permeability). Methyl tert-butyl ether caused more inhibition of alpha-aminoisobutyric acid absorption (64%) than mono-octanoin (48%) and a greater reduction of dry weight per centimeter of the perfused segment (22%) compared with mono-octanoin (10%). Such effects appeared after only 1 min of solvent exposure and did not appreciably increase with longer exposures. Permeation of mannitol increased by 26% after 1 min of exposure to mono-octanoin and by 54% after a similar period of exposure to methyl tert-butyl ether. Longer exposures to both solvents did not seem to cause progressive increases in mannitol permeation. The results indicate that brief exposure of the rat jejunum to either of the two solvents causes a reduction in active transport ([14C]alpha-aminoisobutyric acid absorption), an increase in passive permeability (mannitol permeation), and a loss of mucosal constituents. We conclude that the intestinal mucosa is susceptible to solvent damage and may be used as a selectively sensitive model that can characterize the biological injury of gallstone solvents. The study also suggests that escape of the currently available solvents into the small intestine in patients undergoing contact dissolution of gallbladder stones may cause injury to the small intestine.

Effect of some triazole fungicides on intracellular pH and on cell membrane permeability in leaves of Elodea densa (Planch.) Casp.

Recent data obtained in this laboratory have shown that a number of triazole fungicides induce in leaves of barley and Elodea densa an early release of solutes and an inhibition of H+ extrusion. This paper deals with the early effects of two triazoles, penconazole and flusilazol, on intracellular pH and plasmamembrane permeability in Elodea densa leaves. Different effects were found at low (0.1-0.15 mM) or at high (0.3-0.5 mM) triazole concentrations. At the low concentrations the triazoles induced a marked intracellular acidification (corresponding to an acidification of both the cytosol and the vacuole) as evaluated by the weak acid and weak base distribution method. For higher triazole concentrations (0.3-0.5 mM) the intracellular pH became very close to that of the external medium (pH 5.5), and cytosolic and vacuolar pH measurements became unreliable. Triazoles at concentrations higher than 0.1-0.15 mM induced an early and very marked increase in net efflux of K+ and Cl- . At these 'toxic' concentrations the sorbitol-permeable space (apparent free space) was increased from about 20% to more than 90%. These results suggest that triazoles at low 'sub-toxic' concentrations inhibit some mechanism involved in the regulation of intracellular pH (possibly the plasmalemma proton pump), whilst at higher concentrations they induce a non-specific increase in passive permeability of the plasmamembrane.

Control of membrane permeability by external adenosine triphosphate in animal cells and its application to chemotherapy

External adenosine triphosphate (ATP) has been shown to induce permeability changes in a variety of animal cells. This review is mainly concerned with the effect of external ATP on passive permeability of transformed cells in culture. Brief exposure of several transformed cells to ATP causes a striking increase in passive permeability, allowing passage through the plasma membrane of phosphorylated metabolites and ions. Untransformed fibroblasts do not respond to the ATP. Although the molecular mechanisms of this permeability change in transformed cells are largely unknown, it has been demonstrated that the permeability change is modulated by cellular ATP, cytoskeleton and calmodulin. These characteristics and the recent studies with an isolated ATP-resistant variant designed to gain some insight into the biochemistry and regulation of the ATP effect are summarized. By considering a selective effect of external ATP on membrane permeability of transformed cells, possible application of this method to cancer chemotherapy has been examined. The results obtained mainly with in vitro systems are also described.

Reciprocal effects of phenothiazines and naphthalene sulfonamides on the external ATP-dependent permeability change in Chinese hamster ovary cells.

External ATP causes a great increase in passive permeability to phosphorylated metabolites in several transformed cells, but not in untransformed cells. We have previously demonstrated that the external ATP-dependent permeability change was induced in Chinese hamster ovary cells, CHO-K1, only in the presence of a mitochondrial inhibitor (rotenone, KCN) or a cytoskeleton-attacking agent, vinblastine (Kitagawa, T. and Akamatsu, Y. Biochim. Biophys. Acta 649, 76-82 (1981); 734, 25-32 (1983]. A similar ATP-dependent permeability change was also induced in CHO cells when the cells were treated with 10-30 microM trifluoperazine. This permeability change, like the previously mentioned ones, was found to be reversible and the treated cells remained viable. The permeability change induced by ATP and trifluoperazine was independent of changes in cellular ATP concentration and this property was the same as that of the permeability change with external ATP and vinblastine. Since trifluoperazine is known to interact with calmodulin and to inhibit calmodulin-dependent cellular functions, these results may indicate that calmodulin associated with the cytoskeleton plays an important role in control of the permeability change, although nonspecific perturbation by the drug of the membranes cannot be ruled out. Chlorpromazine and a naphthalene sulfonamide, W-7, also induced an ATP-dependent permeability change. However, these drugs, like mitochondrial inhibitors, reduced the cellular ATP concentration to induce the permeability change. Thus, a clear difference in the action of these drugs in intact cells was also shown in this study. Possible mechanisms for the ATP-dependent permeability change in mammalian cells are discussed.

Decline in Energy Reserves of Chlorella sorokiniana upon Exposure to Ozone

Ozone exposure has been shown to increase the loss of K from Chlorella cells due to an increase in passive permeability and a depolarization of membrane potential. One factor which likely influences or can be influenced by these changes is the energy state of the cell. To study this relationship, various indicators of cell energy status were examined in the presence and absence of O(3).The active uptake of chloride and deoxyglucose is nearly completely inhibited by O(3) at a dose at which cellular death, measured by plating efficiency, is minimal. Glucose-stimulated respiration, dependent upon ATP/ADP balance, is depressed to a greater degree than endogenous respiration in ozonated cells. Total ATP and glucose-6-phosphate levels also decrease but not as rapidly, and labeled intermediates of glucose metabolism are lost.Thus, exposure to O(3) results in a depletion of the cell's energy reserves as substantiated by changes observed in processes which both utilize and generate ATP. This loss in energy reserves occurs at the same exposure level of O(3) as do the changes in passive transport properties. Thus, we cannot tell which occurs first; and the processes seem to be linked with respect to O(3) injury.

The role of calcium ions in the permeability changes produced by external ATP in transformed 3T3 cells

External ATP causes a rapid increase in passive permeability to nucleotides and phosphate esters in transformed cell lines, such as 3T6 mouse fibroblasts. However, untransformed lines, such as 3T3, do not show a similar sensitivity to external ATP. Ca 2+ inhibits permeabilization, but only at concentrations approaching those of external ATP. In contrast, La 3+ and Tb 3+ inhibit ATP-dependent permeabilization at one-fifth the concentration of external ATP. Considering reports that lanthanides can substitute for calcium ion in many enzymatic reactions, often with a higher affinity, it would appear that Ca 2+ plays a specific role in the maintenance of a passive membrane permeability barrier and in opposing the effects of external ATP. Other data suggest a regulatory role for the Ca 2+-calmodulin complex in the permeabilization process. Trifluoperazine, chlorpromazine and W-7, compounds which inhibit cellular functions dependent on the Ca 2+-calmodulin complex, are able to enhance the effect of external ATP. Thus, a dramatic stimulation of nucleotide permeability occurs with concentrations of external ATP and inhibitor that are ineffective when added alone. Calmodulin antagonists and low concentrations of external ATP increased membrane permeability to Na + and K + as was previously shown for permeabilization with ATP alone. Earlier studies have shown that energy inhibitors which reduce intracellular ATP levels greatly increase the sensitivity of transformed cells to external ATP. However, the Ca 2+-calmodulin antagonists used in the present study exert their effects at concentrations which do not alter intracellular ATP levels.

Proximal tubular HCO3-, H+ and fluid transport during maleate-induced acidification defect.

The mechanism of tubular acidification was studied in proximal tubular acidification defect induced in rats by acute parenteral infusion of maleate (200 mg/kg), which causes diuresis and bicarbonaturia. Proximal tubular bicarbonate reabsorption and H+ ion secretion were determined by stopped-flow microperfusion and measurement of luminal pH by Sb microelectrodes. Stationary pH increased in proximal tubule from 6.78 to 7.25 and bicarbonate reabsorption decreased from 1.32 to 0.51 nmol/cm2 X s. In these segments, mean cell PD fell from -66.6 to -20.2 mV, while Jv as estimated by the Gertz technique fell to 15% of controls. A similar impairment of acidification was observed during luminal and capillary perfusion with phosphate Ringer's. Since H+-ion efflux from the lumen was not significantly increased and both acidification and alkalinization half-times (t/2) were increased, no evidence for an increase in passive permeability for H+/HCO3- was obtained. The increased t/2 found during luminal perfusion with acid phosphate indicates, according to an electrical analog model, a reduction in pump series conductance. These results show that maleate affects both proximal Na+ and H+ transport; this effect may be ascribed to impairment of sodium-dependent transport systems in the brush-border membrane.

Reduced ocular glucose transport and increased non-electrolyte permeability in rats with retinal degeneration (RCS)

The transport kinetics across the plasma-ocular barriers of labeled molecules including urea, d-glucose, 3-0-methyl- d-glucose, l-glucose, and sucrose were studied in adult RCS rats and compared to control albino Sprague-Dawleys (SD). Results indicate that substances that passively cross the plasma-aqueous and plasma-vitreous barriers (urea, sucrose, l-glucose) do so more readily in the RCS rat especially via the trans-retinal route. By contrast, d-glucose, which penetrates the ocular barriers of the control rat by a carrier-mediated mechanism was found to cross the ocular barriers of the RCS rat at a reduced rate. Thus, the ocular barriers of the RCS rat with well developed retinal degeneration demonstrate an increase in passive permeability and a reduction in ability to perform their membrane transport function ( d-glucose). The permeability defect was more pronounced in the blood-vitreous barrier than in the blood-aqueous barrier.

Characterization of free radical-mediated damage of canine cardiac sarcoplasmic reticulum

In vitro generation of free radicals by xanthine oxidase acting on xanthine as substrate depressed steady-state calcium uptake by canine cardiac sarcoplasmic reticulum vesicles. The effect of free radicals on the calcium-dependent ATPase activity of the SR vesicles was pH dependent. At pH 7.0, ATPase activity was decreased, but at pH 6.4 it was unchanged. Exposure to free radicals increased the passive permeability of the vesicles to calcium. This increase was inhibited by superoxide dismutase (SOD) at pH 7.0, and SOD plus mannitol at pH 6.4. The increased permeability per se was insufficient to explain the effects of free radicals on ATPase activity, since the calcium ionophore A23187 was unable to mimick these effects. Direct measurement of the number and turnover of the pump units indicated that the number of units was unchanged but turnover was decreased by free radicals at pH 7.0. The overall data suggest at least two mechanisms of free radical damage, one associated with an increase in passive permeability and another associated with an as yet undefined change in some specific steps of the ATPase reaction.

Characterization of free radical-mediated damage of canine sarcoplasmic reticulum.

In vitro generation of free radicals by xanthine oxidase acting on xanthine as a substrate depressed steady-state calcium uptake by canine cardiac sarcoplasmic reticulum (SR) vesicles. The effect of free radicals on the calcium-dependent ATPase activity of the SR vesicles depended on pH. At pH 7.0, ATPase activity was decreased, but at pH 6.4»it was unchanged. Exposure to free radicals increased the passive permeability of the vesicles to calcium. This increase was reversible by superoxide dismutase (SOD) at pH 7.0 and by SOD plus mannitol at pH 6.4. The increased permeability per se was insufficient to explain the effects of free radicals on ATPase activity since the calcium ionophore A23187 was unable to mimick these effects. Direct measurement of the number and turnover of the pump units indicated that the number of units is unchanged, but turnover is decreased by free radicals at pH 7.0. The overall data suggest there are at least two mechanisms of free radicals damage, one associated with an increase in passive permeability and another associated with an as yet undefined change in some specific steps of the ATPase reaction. The reversibility of the damage produced by the free radicals is a potentially extremely important finding. It clearly demonstrates that free radical-mediated damage to tissues might be reversible if effective intervention takes place shortly after damage occurs.

The effect of in vitro and in vivo lead intoxication on monosaccharide transport in isolated rat brain microvessels

The effect of lead on facilitated transport of monosaccharides into isolated brain microvessels was investigated. Preparations from control rats and from rats exposed to lead acetate at various doses (0.025–1.0 mg lead/g body wt/day) from days 5–25 postnatally were studied. The time courses of uptake for various concentrations of 3-O-methylglucose (3OMG) were determined. Values of 0.6 min and 2.0 min for 5 and 50 mM 3OMG, respectively, were obtained in the case of preparations from control animals. In contrast, the half-time of uptake was about 1 min for both sugar concentrations for microvessels isolated from animals treated at 1.0 mg lead/g body wt/day. This indicates both a block in facilitated transport of the sugar, as well as damage to the endothelial cells resulting in increased passive permeability. There was a dose-response relationship for the effect of in vivo lead treatment on 3OMG transport in isolated microvessels. Microvessel preparations from younger animals (treated at 1.0 mg/g body wt for two days prior to sacrifice at 7 days postnatally) were more sensitive to lead treatment than preparations from adults; equivalent lead burden induced a greater increase in passive permeability to 3OMG in the younger animals. In other experiments microvessels isolated from untreated rats were preincubated with lead acetate. A concentration of 0.1 μM lead acetate in vitro abolished facilitated transport of 3OMG.