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S R Ennis

Publications and source records attributed to S R Ennis.

16 recordsLinked to original sources
PubMed1996

Mechanisms of sodium transport at the blood-brain barrier studied with in situ perfusion of rat brain.

The mechanism of unidirectional transport of sodium from blood to brain in pentobarbital-anesthetized rats was examined using in situ perfusion. Sodium transport followed Michaelis-Menten saturation kinetics with a Vmax of 50.1 nmol/g/min and a Km of 17.7 mM in the left frontal cortex. The kinetic analysis indicated that, at a physiologic sodium concentration, approximately 26% of sodium transport at the blood-brain barrier (BBB) was carrier mediated. Dimethylamiloride (25 microM), an inhibitor of Na+/H+ exchange, reduced sodium transport by 28%, whereas phenamil (25 microM), a sodium channel inhibitor, reduced the transfer constant for sodium by 22%. Bumetanide (250 microM) and hydrochlorothiazide (1.5 mM), inhibitors of Na(+)-K(+)-2Cl-/NaCl symport, were ineffective in reducing blood to brain sodium transport. Acetazolamide (0.25 mM), an inhibitor of carbonic anhydrase, did not change sodium transport at the BBB. Finally, a perfusate pH of 7.0 or 7.8 or a perfusate PCO2 of 86 mm Hg failed to change sodium transport. These results indicate that 50% of transcellular transport of sodium from blood to brain occurs through Na+/H+ exchange and a sodium channel in the luminal membrane of the BBB. We propose that the sodium transport systems at the luminal membrane of the BBB, in conjunction with Cl-/HCO3- exchange, lead to net NaCl secretion and obligate water transport into the brain.

Amiloride↗
PubMed1995

Developmental changes in blood-brain barrier potassium permeability in the rat: relation to brain growth.

1. The potassium permeability of the blood-brain barrier (BBB) was determined in anaesthetized rats aged between 21 days gestation and adult using 86Rb+ as a marker for potassium. 2. The brain influx rate constant for 86Rb+ was high in fetal cortex at 21 days gestation (42.5 +/- 4.3 microliters g-1 min-1) but had decreased markedly by just after birth (12.2 +/- 0.6 microliters g-1 min-1). There was a further, gradual, postnatal decline to 7.0 +/- 0.3 microliters g-1 min-1 by 50 days after birth. 3. Developmental changes in passive BBB permeability were examined over the same age range using 14[C]urea. These studies showed similar developmental changes in influx rate to those found for 86Rb+. Specifically, a marked perinatal decline followed by a more gradual postnatal fall. Thus, the changes in potassium permeability probably reflect a decrease in the BBB paracellular leak during development. 4. The changes in BBB permeability coincide with changes in the rate of brain growth and the associated rate of brain potassium accumulation. As the potassium permeability properties of the adult BBB would provide insufficient potassium influx to meet the requirement associated with fetal brain growth, it is suggested that need for potassium may be the reason for the greater BBB permeability early in development.

Animals↗
PubMed1994

Transport of alpha-aminoisobutyric acid across the blood-brain barrier studied with in situ perfusion of rat brain.

Transport of alpha-aminoisobutyric acid (AIB) from blood to brain in pentobarbital-anesthetized rats was examined using in situ perfusion. In situ perfusion with washed sheep red blood cells allowed the precise control of the composition of the perfusate that was necessary for a detailed examination of the transport of AIB. Retrograde perfusion at 4 ml/min through the left external carotid artery with oxygenated, artificial blood (hematocrit = 0.3) maintained a normal electroencephelogram during a 10 min experiment. The perfusate cerebral blood flow, at a value of 1.2 +/- 0.1 ml/g/min, and the perfusate cerebral plasma volume, at a value of 5.4 +/- 1.9 microliter/g, in the left frontal cortex were within the range of reported in vivo values. The in situ PS product for AIB (3.8 +/- 0.4 microliter/g/min) was higher than the value observed in vivo. AIB uptake was reduced to the in vivo value by 2 mM phenylalanine (1.3 +/- 0.3 microliter/g/min) and equally well by a mixture of neutral amino acids at their normal plasma concentrations but was unaffected by 2 mM methyl-AIB or removal of sodium from the perfusate. A kinetic analysis showed that the apparent Ki for phenylalanine inhibition of AIB transport was 19.8 +/- 4.9 microM. Thus, although AIB has affinity for the large neutral amino acid carrier in the blood-brain barrier, brain uptake by this mechanism in vivo is negligible due to competition by other amino acids in the plasma.

Amino Acids↗
PubMed1994

Riboflavin reduces edema in focal cerebral ischemia.

Oxidized irwN has been proposed as a mediator of the free radical-induced damage that occurs during cerebral ischemia. Dihydroriboflavin, a compound produced from riboflavin (B2) by NADPH-dependent flavin reductase, rapidly reduces oxidized iron. Since treatment with riboflavin offers protection from ischemic injury in other tissues, we tested the effect of pretreatment with B2 on brain edema formation during focal ischemia. Two different models of middle cerebral artery occlusion (MCAO) in rats were tested: transcranial electrocautery and intracarotid occlusion with a nylon thread. Groups of 6-8 animals were treated with 7.5 mg of B2/kg or saline vehicle 1 h before MCAO and brain water content was determined after 4 h of ischemia. Pretreatment with B2 reduced total hemisphere edema formation from 0.37 +/- 0.05 to 0.19 +/- 0.05 mg/g dry wt. (48% protection, p < 0.01) following transcranial MCAO. Edema was greater following MCAO with the intra-carotid thread (0.54 +/- 0.05 ml/g) but protection by B2 was less (21%). We conclude that pretreatment with B2 reduces ischemic brain injury, perhaps by reacting with oxidized iron. However, the larger stroke produced by the thread MCAO method makes it more difficult to observe protection following brief ischemia in this model.

Animals↗
PubMed1993

Evidence that NADPH-dependent methemoglobin reductase and administered riboflavin protect tissues from oxidative injury.

NADPH-dependent methemoglobin reductase, first detected in erythrocytes sixty years ago, has subsequently been purified and characterized as a methylene blue reductase and a flavin reductase. The reductase plays no role in methemoglobin reduction under normal conditions, but its activity serves as the basis for the treatment of methemoglobinemia with methylene blue or flavin. On-going studies demonstrate that this cytosolic protein is also present in liver and that its primary structure distinguishes it from other known proteins. The bovine erythrocyte reductase tightly binds hemes, porphyrins, and fatty acids with resulting loss of activity. Pyrroloquinoline quinone serves as a high-affinity substrate of the reductase, suggesting that this naturally-occurring compound may be a physiological substrate. The ability of the reductase to catalyze the intracellular reduction of administered riboflavin to dihydroriboflavin suggested that this system might be exploited to protect tissues from oxidative damage. This hypothesis was supported by our finding that dihydroriboflavin reacts rapidly with Fe(IV)O and Fe(V)O oxidation states of hemeproteins, states that have been implicated in tissue damage associated with ischemia and reperfusion. Preliminary studies demonstrate that, as predicted, administration of low concentrations of riboflavin protects isolated rabbit heart from reoxygenation injury, rat lung from injury resulting from systemic activation of complement, and rat brain from damage caused by four hours of ischemia. Data from these animal studies suggest that flavin therapy holds promise in protecting tissue from the oxidative injuries of myocardial infarction, acute lung injury, stroke, and a number of other clinical conditions.

Animals↗
PubMed1990

Decrease in perfusion of cerebral capillaries during incomplete ischemia and reperfusion.

The effect of unilateral, incomplete cerebral ischemia on CBF, unidirectional flux of alpha-aminoisobutyric acid (AIB) and sodium, and number of perfused capillaries during ischemia and reperfusion was measured in the cortex of gerbils with symptomatic ischemia. Three hours of unilateral carotid occlusion reduced the CBF to the ipsilateral cortex by 81%, with a smaller 30% decrease in the contralateral cortex. Following 11 min of reperfusion, CBF in the ipsilateral cortex returned to the preischemic value, while the contralateral blood flow decreased to 50% of control. The transfer constants for AIB and sodium in the ipsilateral cortex were reduced by 67 and 53%, respectively, after 3 h of ischemia, with no change in the contralateral cortex. The transfer constant for AIB remained decreased by 48% during the first 20 min of reperfusion, while that for sodium returned to its control value. The number of perfused capillaries was reduced 54% by 3 h of ischemia and remained decreased by 20% after 11 min of reperfusion. These data indicate that 3 h of unilateral carotid occlusion reduces the number of perfused capillaries in the ipsilateral cortex during the ischemic period. Further, the early reperfusion phase is characterized by a mismatch between capillary perfusion and CBF. Finally, early in the postischemic phase, sodium transport undergoes a selective stimulation, probably as a result of stimulation of ion transport.

Aminoisobutyric Acids↗
PubMed1990

Permeability of the blood-ocular barrier to mannitol and PAH during experimental diabetes.

The simultaneous uptake of mannitol, a passive permeability marker, and the organic acid, p-aminohippuric acid, was measured in the anterior and posterior vitreous and retina. Uptake was determined in control, probenecid treated and streptozocin diabetic rats. The unidirectional influx, calculated as a 10 minute PS product for these compounds, was not increased by 2, 4, or 12 weeks of diabetes. A clearance value, calculated using an experimental time of 60 minutes was also examined in order to gauge efflux of these compounds from the eye. The 60 minute clearance value for mannitol in the retina increased approximately 75% in 2, 4, and 12 week diabetic rats. This was an unexpected result due to the lack of increase in the unidirectional flux of mannitol during these same periods of diabetes, and may represent a change in the passive efflux out of the retina. The 60 minute clearance value for mannitol was not significantly changed in either the anterior or posterior vitreous. Experimental diabetes increased the 60 minute clearance value for PAH for the retina by 40% to 70%. In contrast, diabetes did not increase influx of PAH into the anterior or posterior vitreous. Because the unidirectional influx of PAH into the retina was not increased during diabetes, a decrease in the active transport for organic acids out of the eye is a likely explanation for the increase in the 60 minute clearance value for PAH during diabetes.

Analysis of Variance↗
PubMed1990

Blood-to-brain sodium transport in ischemic brain edema.

Brain edema is a frequent complication of cerebral ischemia; however, its mechanism of formation is not well understood. Sodium is known to accumulate in brain during the early stages of partial ischemia. Therefore, the present studies were undertaken to determine the relation among BBB sodium transport, integrity of the BBB, and development of brain edema during the first 24 hr after the onset of cerebral ischemia. Partial cerebral ischemia was produced in gerbils by ligation of the left common carotid artery under ether anesthesia. After recovery from the anesthetic, animals were scored for the presence of symptoms, and those with scores greater than 10 of 25 (n = 87) were chosen for this study. Measurements of tissue water, sodium, and potassium contents, and brain uptake of 22Na and 3H-mannitol were made in each group at 1.5, 3, 6, 12, and 24 hr after carotid ligation. Accumulation of sodium and water in the ischemic compared with the nonischemic cerebral cortex was progressive. This edema formation was not of the vasogenic type because the permeability of the BBB to mannitol was unchanged. Blood-to-brain sodium transport was reduced by 30% to 40% at all time points in the ischemic cortex. Nevertheless, the remaining sodium transport activity appeared to play a role in the development of brain edema because Na accumulated in the tissue at a rate that was approximately the same as the rate of 22Na uptake from blood.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Blood-brain barrier sodium transport limits development of brain edema during partial ischemia in gerbils.

Sodium derived from the blood is known to accumulate in brain tissue during the early stages of incomplete ischemia. Our present studies were undertaken to determine the relation between blood-brain barrier sodium transport and the development of ischemic brain edema. Incomplete cerebral ischemia was produced in gerbils by ligation of the left common carotid artery under ether anesthesia. Following recovery from the anesthetic, the gerbis were evaluated for the presence of neurologic symptoms and were divided into symptomatic (n = 77) and asymptomatic (n = 94) groups. Tissue water, sodium, and potassium contents, tissue plasma volume, and brain uptake of 22Na were measured in both groups 1.5, 3, 6, 12, and 24 hours after carotid ligation. There was a progressive accumulation of sodium and water in the ipsilateral cerebral cortex of the symptomatic group compared with either the corresponding contralateral cortex of the same gerbils or with the asymptomatic group. Net changes in brain sodium and potassium concentrations appeared to be the main determinants of fluid accumulation. Brain edema was not due to opening of the blood-brain barrier because the unidirectional transport of 22Na remained low and was even reduced by 35-55% in the ischemic cortex. Nevertheless, this sodium transport activity appeared to be rate-limiting in the development of brain edema during the first 3 hours of ischemia because the rate of sodium accumulation in the tissue was the same as the rate of 22Na transport from the blood to the brain. We conclude that blood-brain barrier sodium transport is an important factor in the formation of ischemic brain edema.

Animals↗
PubMed1986

Sucrose permeability of the blood-retinal and blood-brain barriers. Effects of diabetes, hypertonicity, and iodate.

The permeabilities of the blood-retinal (BRB) and blood-brain (BBB) barriers to sucrose were determined simultaneously using an intravenous injection technique in the rat. The method involved direct sampling of retinal tissue in order to avoid errors caused by sucrose penetration across other components of the blood-ocular barrier. The permeability X surface area (PS) product for the BRB was approximately four times greater than for the BBB. Intracarotid infusion of a hypertonic arabinose solution resulted in a dose-dependent increase in the permeability of both barrier systems. In contrast, 24 hr after treatment of animals with iodate, the PS product for the BRB but not the BBB was increased. The permeability of the blood-retinal barrier to sucrose was measured in normal and 2-, 6-, and 20-week streptozocin diabetic rats. The BRB was unaffected at 2 and 6 weeks of diabetes, and showed only a small increase in permeability at 20 weeks. Our results suggest that alterations in the blood-ocular barrier in early diabetes do not result from an increased passive permeability of the BRB. The method described should permit direct comparison of BRB and BBB permeabilities to a variety of compounds under various conditions.

Animals↗
PubMed1984

The effect of diet on inherited retinal dystrophy in the rat.

The rate of photoreceptor cell degeneration was significantly retarded in albino rats afflicted with inherited retinal dystrophy by merely changing the food supply from a commercial lab chow to a well defined diet. The data suggest that female dystrophic rats may benefit somewhat more from this dietary manipulation than the males. A well defined fat-free diet was less successful than a standard defined diet in retarding the rate of retinal degeneration.

Animals↗
PubMed1983

Brain microvessel endothelial cells in tissue culture: a model for study of blood-brain barrier permeability.

Endothelial cells were prepared from bovine microvessels and grown in tissue culture. They contained factor VIII/von Willebrand antigen, the most specific market available for determination of the endothelial origin of cells in culture. The cultured cells formed complex tight junctions and contained few pinocytotic vessels. These properties are responsible for formation of the blood-brain barrier in vivo. When monolayers of the endothelial cells were exposed briefly to a calcium-free solution or treated with 1.6 M arabinose, distinctive morphological changes occurred in the intercellular contacts. In either case, a normal structure was reestablished following return to control medium. To assess the effect of these treatments on transcellular permeability, we measured the movement of sucrose labeled with carbon 14 across a monolayer of endothelial cells cultured on a collagen-coated nylon mesh. Removal of external calcium increased the rate of sucrose movement by 120%; the arabinose treatment increased transcellular flux by 40%.

Animals↗
PubMed1982

In situ kinetics of glucose transport across the blood-retinal barrier in normal rats and rats with streptozocin-induced diabetes.

Glucose transport across the blood-retinal barrier (BRB) was investigated in situ by means of a modification of the single-injection technique developed for studying transport across the blood-brain barrier. A retinal uptake index was calculated from the fractional extraction for 14C-glucose divided by the fractional extraction for the 3H2O reference. The use of the Eadie-Hofstee transformation to determine the Km of carrier-mediated transport revealed at least two transport systems for the unidirectional glucose flux across the BRB. Control data showed a relatively high-affinity system with a Km of 0.24 mM and a lower affinity system with a Km of 7.81 mM. Both these transport systems have been shown to be sensitive to phloretin but not to phloridzin or acetazolamide. It has also been shown that glucose transport is uneffected by Na+ or K+ concentrations but that calcium does have a significant effect on the lower affinity transport system for glucose. We have also demonstrated that the Km values of both transport systems are significantly increased by 2 months of streptozocin-induced diabetes.

Acetazolamide↗
PubMed1980

The effect of induced diabetes on the electroretinogram components of the pigmented rat.

The fast and slow components of the electroretinogram (ERG) of the pigmented rat were observed 2, 4, and 19 weeks after the induction of diabetes by streptozotocin. The diabetic state was confirmed by measures of serum and urine glucose and the appearance of polydipsia and polyuria. There was no apparent effect on the b-wave of the ERG in the diabetic animals at the 2- and 4-week periods of observation; however, at 19 weeks the form of the intensity-amplitude curve differed from that of the controls. The c-wave of the ERG was significantly reduced in amplitude at the 2-week period of observation, and a progressive reduction occurred throughout the remaining periods of observation. The relationship between the observed reduction in amplitude of the c-wave and the reported pathological changes in the pigment epithelium in diabetic animals is discussed.

Animals↗
PubMedDate not supplied

The effects of galactosamine-induced hepatic failure upon blood-brain barrier permeability.

The role of changes in blood-brain barrier permeability in the pathogenesis of hepatic encephalopathy remains uncertain. To test the hypothesis that brain microvessel permeability is nonselectively increased in hepatic encephalopathy we measured the blood-brain barrier permeability-surface area product in rats with acute liver failure induced by intraperitoneal injection of galactosamine. The permeability-surface area products to the diffusion-limited tracers, sucrose and methylaminoisobutyric acid, were determined as a measure of blood-brain barrier permeability. Animals were examined 24, 36 and 42 hr after injection, at times when they were stuporous, but not comatose. No significant elevations of the permeability-surface area products for either compound were detected in clinically affected experimental animals when compared to controls. Our results indicate there is no generalized increase in brain vascular permeability during hepatic insufficiency in precomatose animals.

Animals↗