Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose... more Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose or L-fucose concentrations. Chronic exposure of neuroblastoma cells to 30 mM glucose or 30 mM L-fucose caused a decrease in ouabain-sensitive and veratridine-stimulated 22Na+ uptake compared with cells cultured in unsupplemented medium. The Na+ current, determined by using whole-cell configuration of the patch clamp, was also decreased in these cells. Tetrodotoxin (3 microM), which blocked whole cell Na+ currents, also blocked veratridine-stimulated 22Na+ accumulation. Culturing cells in medium containing 30 mM fructose as an osmotic control had no effect on Na+ flux. Specific [3H]saxitoxin binding was not affected by 30 mM glucose or 30 mM L-fucose compared with cells grown in unsupplemented medium, suggesting that the number of Na+ channels was not decreased. These studies suggest that exposing cultured neuronal cells to conditions that occur in the diabetic milieu alters Na+ transport and Na(+)-channel activity.
Metabotropic glutamate (mGlu) receptors provide a mechanism by which the function of NMDA glutama... more Metabotropic glutamate (mGlu) receptors provide a mechanism by which the function of NMDA glutamate receptors can be modulated. As NMDA receptor hypofunction is implicated in the etiology of psychiatric disorders, including schizophrenia, the pharmacological regulation of mGlu receptor activity represents a promising therapeutic approach. We examined the effects of the positive allosteric mGlu(5) receptor modulator 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide (CDPPB), alone and in combination with the NMDA receptor antagonist MK-801, on a task measuring cognitive set-shifting ability. This task measures NMDA receptor-dependent cognitive abilities analogous to those impaired in schizophrenia. Systemic administration of CDPPB (10 and 30 mg/kg i.p) blocked MK-801 (0.1mg/kg, i.p.)-induced impairments in set-shifting ability. The effect on learning was dose-dependent, with the 30 mg/kg dose having a greater effect than the 10mg/kg dose across all trials. This ameliorative effect of ...
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology, 2004
Pharmacological manipulation of N-methyl-D-aspartate (NMDA) receptors may be critical for the tre... more Pharmacological manipulation of N-methyl-D-aspartate (NMDA) receptors may be critical for the treatment of many neurological and psychiatric disorders. Metabotropic glutamate (mGlu5) receptors are abundant in corticolimbic circuitry, where they modulate NMDA receptor-mediated signal transduction. Therefore, pharmacological manipulation of mGlu5 receptor may provide a treatment strategy for cognitive disorders that are associated with NMDA receptor dysfunction. We sought to determine whether the recently described molecular and cellular interactions between NMDA and mGlu5 receptors coregulate higher order behaviors. We examined the interaction of the selective mGlu5 receptor antagonist, 2-methyl-6-(phenylethynyl)-pyridine (MPEP), and the use-dependent NMDA antagonist MK-801, on locomotion, stereotypy, working memory, instrumental learning, and corticolimbic dopamine release. MPEP, at 10 mg/kg, but not 3 mg/kg, impaired working memory and instrumental learning, transiently increased d...
myo-Inositol accumulation and incorporation into phosphoinositides was decreased in neuroblastoma... more myo-Inositol accumulation and incorporation into phosphoinositides was decreased in neuroblastoma cells chronically exposed to medium containing 30 mmol/L glucose or 30 mmol/L galactose. In addition, the intracellular content of myo-inositol and phosphatidylinositol was decreased and the sorbitol or galactitol content increased in cells cultured for 2 weeks in medium containing 30 mmol/L glucose or 30 mmol/L galactose, respectively. Na+/K+ adenosine triphosphatase (ATPase) transport activity was also significantly decreased by long-term exposure of neuroblastoma cells to medium containing 30 mmol/L glucose or 30 mmol/L galactose. When glucose-conditioned cells were placed in medium containing a normal glucose concentration for 24 hours, myo-inositol metabolism and content, phosphatidylinositol levels, and Na+/K+ pump activity were restored or completely returned to normal values. These functions were also significantly improved, except for the phosphatidylinositol content, which was increased by 55%, when galactose-conditioned cells were incubated for 24 hours in unsupplemented medium. The polyol content of the glucose- or galactose-conditioned cells was also significantly reduced. Returning the cells to normal glucose levels for 1 to 3 hours did not completely restore myo-inositol metabolism. Improved myo-inositol metabolism and content, sorbitol levels, and Na+/K+ ATPase transport activity were also obtained within 24 hours when cells chronically exposed to medium supplemented with 30 mmol/L glucose were placed in medium containing 30 mmol/L glucose and 0.4 mmol/L sorbinil. The phosphatidylinositol content of these cells was improved by approximately 30%. Cells prelabeled for 24 hours with [U-14C]sorbitol metabolize more than 50% of the [U-14C]sorbitol during a 24-hour incubation in unsupplemented medium. These studies conducted at the cellular level suggest that restoration of normal myo-inositol metabolism, polyol content, and Na+/K+ pump activity altered by hyperglycemic conditions occurs rapidly following normalization of glucose concentration.
In population learning studies, between-subject response differences are an important source of v... more In population learning studies, between-subject response differences are an important source of variance that must be characterized to identify accurately the features of the learning process common to the population. Although learning is a dynamic process, current population analyses do not use dynamic estimation methods, do not compute both population and individual learning curves, and use learning criteria that are less than optimal. We develop a state-space random effects (SSRE) model to estimate population and individual learning curves, ideal observer curves, and learning trials, and to make dynamic assessments of learning between two populations and within the same population that avoid multiple hypothesis tests. In an 80-trial study of an NMDA antagonist's effect on the ability of rats to execute a set-shift task, our dynamic assessments of learning demonstrated that both the treatment and control groups learned, yet, by trial 35, the treatment group learning was significantly impaired relative to control. We used our SSRE model in a theoretical study to evaluate the design efficiency of learning experiments in terms of the number of animals per group and number of trials per animal required to characterize learning differences between two populations. Our results demonstrated that a maximum difference in the probability of a correct response between the treatment and control group learning curves of 0.07 (0.20) would require 15 to 20 (5 to 7) animals per group in an 80 (60)-trial experiment. The SSRE model offers a practical approach to dynamic analysis of population learning and a theoretical framework for optimal design of learning experiments.
Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose... more Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose or L-fucose concentrations. Chronic exposure of neuroblastoma cells to 30 mM glucose or 30 mM L-fucose caused a decrease in ouabain-sensitive and veratridine-stimulated 22Na+ uptake compared with cells cultured in unsupplemented medium. The Na+ current, determined by using whole-cell configuration of the patch clamp, was also decreased in these cells. Tetrodotoxin (3 microM), which blocked whole cell Na+ currents, also blocked veratridine-stimulated 22Na+ accumulation. Culturing cells in medium containing 30 mM fructose as an osmotic control had no effect on Na+ flux. Specific [3H]saxitoxin binding was not affected by 30 mM glucose or 30 mM L-fucose compared with cells grown in unsupplemented medium, suggesting that the number of Na+ channels was not decreased. These studies suggest that exposing cultured neuronal cells to conditions that occur in the diabetic milieu alters Na+ transport and Na(+)-channel activity.
Cultured neuroblastoma, cerebral microvessel endothelial, and retinoblastoma cells were used to e... more Cultured neuroblastoma, cerebral microvessel endothelial, and retinoblastoma cells were used to examine the mechanism of acute inhibition by D-glucose of myo-inositol uptake. Acute exposure of the cells to 30 mM D-glucose caused a significant decrease in Na(+)-dependent myo-inositol uptake in all three cell types. The effect of D-glucose to acutely inhibit myo-inositol uptake was dependent on the extracellular glucose concentration and was not reversed by sorbinil. 2-Deoxy-D-glucose (30 mM), 3-O-methyl-D-glucose (30 mM), and cytochalasin B (100 microM) did not acutely inhibit myo-inositol uptake. These data suggest that the hydroxyl groups on carbons 2 and 3 of D-glucose, which in a Haworth projection appear trans to each other, are important for inhibitory activity. Other monosaccharides (30 mM) having a similar 2,3-trans-diol configuration, L-glucose, D- and L-fucose, D- and L-galactose, D- and L-xylose, and D-arabinose, all to varying degrees significantly inhibited myo-inositol uptake. In all cases, the L-isomers were more potent inhibitors of myo-inositol uptake than the corresponding D-isomers. Monosaccharides (30 mM) having hydroxyl groups on carbons 2 and 3 in a cis configuration, D-mannose, L-rhamnose, D-allose, and D-ribose, did not acutely inhibit myo-inositol uptake. Replacing the hydroxyl group with a fluorine on carbons 2 or 3 of D-glucose negated its inhibitory activity of myo-inositol uptake. In contrast, replacing the hydroxyl group with a fluorine on carbon 6 of D-glucose did not block its inhibition of myo-inositol uptake.(ABSTRACT TRUNCATED AT 250 WORDS)
Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose... more Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose or L-fucose concentrations. Chronic exposure of neuroblastoma cells to 30 mM glucose or 30 mM L-fucose caused a decrease in ouabain-sensitive and veratridine-stimulated 22Na+ uptake compared with cells cultured in unsupplemented medium. The Na+ current, determined by using whole-cell configuration of the patch clamp, was also decreased in these cells. Tetrodotoxin (3 microM), which blocked whole cell Na+ currents, also blocked veratridine-stimulated 22Na+ accumulation. Culturing cells in medium containing 30 mM fructose as an osmotic control had no effect on Na+ flux. Specific [3H]saxitoxin binding was not affected by 30 mM glucose or 30 mM L-fucose compared with cells grown in unsupplemented medium, suggesting that the number of Na+ channels was not decreased. These studies suggest that exposing cultured neuronal cells to conditions that occur in the diabetic milieu alters Na+ transport and Na(+)-channel activity.
Metabotropic glutamate (mGlu) receptors provide a mechanism by which the function of NMDA glutama... more Metabotropic glutamate (mGlu) receptors provide a mechanism by which the function of NMDA glutamate receptors can be modulated. As NMDA receptor hypofunction is implicated in the etiology of psychiatric disorders, including schizophrenia, the pharmacological regulation of mGlu receptor activity represents a promising therapeutic approach. We examined the effects of the positive allosteric mGlu(5) receptor modulator 3-cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide (CDPPB), alone and in combination with the NMDA receptor antagonist MK-801, on a task measuring cognitive set-shifting ability. This task measures NMDA receptor-dependent cognitive abilities analogous to those impaired in schizophrenia. Systemic administration of CDPPB (10 and 30 mg/kg i.p) blocked MK-801 (0.1mg/kg, i.p.)-induced impairments in set-shifting ability. The effect on learning was dose-dependent, with the 30 mg/kg dose having a greater effect than the 10mg/kg dose across all trials. This ameliorative effect of ...
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology, 2004
Pharmacological manipulation of N-methyl-D-aspartate (NMDA) receptors may be critical for the tre... more Pharmacological manipulation of N-methyl-D-aspartate (NMDA) receptors may be critical for the treatment of many neurological and psychiatric disorders. Metabotropic glutamate (mGlu5) receptors are abundant in corticolimbic circuitry, where they modulate NMDA receptor-mediated signal transduction. Therefore, pharmacological manipulation of mGlu5 receptor may provide a treatment strategy for cognitive disorders that are associated with NMDA receptor dysfunction. We sought to determine whether the recently described molecular and cellular interactions between NMDA and mGlu5 receptors coregulate higher order behaviors. We examined the interaction of the selective mGlu5 receptor antagonist, 2-methyl-6-(phenylethynyl)-pyridine (MPEP), and the use-dependent NMDA antagonist MK-801, on locomotion, stereotypy, working memory, instrumental learning, and corticolimbic dopamine release. MPEP, at 10 mg/kg, but not 3 mg/kg, impaired working memory and instrumental learning, transiently increased d...
myo-Inositol accumulation and incorporation into phosphoinositides was decreased in neuroblastoma... more myo-Inositol accumulation and incorporation into phosphoinositides was decreased in neuroblastoma cells chronically exposed to medium containing 30 mmol/L glucose or 30 mmol/L galactose. In addition, the intracellular content of myo-inositol and phosphatidylinositol was decreased and the sorbitol or galactitol content increased in cells cultured for 2 weeks in medium containing 30 mmol/L glucose or 30 mmol/L galactose, respectively. Na+/K+ adenosine triphosphatase (ATPase) transport activity was also significantly decreased by long-term exposure of neuroblastoma cells to medium containing 30 mmol/L glucose or 30 mmol/L galactose. When glucose-conditioned cells were placed in medium containing a normal glucose concentration for 24 hours, myo-inositol metabolism and content, phosphatidylinositol levels, and Na+/K+ pump activity were restored or completely returned to normal values. These functions were also significantly improved, except for the phosphatidylinositol content, which was increased by 55%, when galactose-conditioned cells were incubated for 24 hours in unsupplemented medium. The polyol content of the glucose- or galactose-conditioned cells was also significantly reduced. Returning the cells to normal glucose levels for 1 to 3 hours did not completely restore myo-inositol metabolism. Improved myo-inositol metabolism and content, sorbitol levels, and Na+/K+ ATPase transport activity were also obtained within 24 hours when cells chronically exposed to medium supplemented with 30 mmol/L glucose were placed in medium containing 30 mmol/L glucose and 0.4 mmol/L sorbinil. The phosphatidylinositol content of these cells was improved by approximately 30%. Cells prelabeled for 24 hours with [U-14C]sorbitol metabolize more than 50% of the [U-14C]sorbitol during a 24-hour incubation in unsupplemented medium. These studies conducted at the cellular level suggest that restoration of normal myo-inositol metabolism, polyol content, and Na+/K+ pump activity altered by hyperglycemic conditions occurs rapidly following normalization of glucose concentration.
In population learning studies, between-subject response differences are an important source of v... more In population learning studies, between-subject response differences are an important source of variance that must be characterized to identify accurately the features of the learning process common to the population. Although learning is a dynamic process, current population analyses do not use dynamic estimation methods, do not compute both population and individual learning curves, and use learning criteria that are less than optimal. We develop a state-space random effects (SSRE) model to estimate population and individual learning curves, ideal observer curves, and learning trials, and to make dynamic assessments of learning between two populations and within the same population that avoid multiple hypothesis tests. In an 80-trial study of an NMDA antagonist's effect on the ability of rats to execute a set-shift task, our dynamic assessments of learning demonstrated that both the treatment and control groups learned, yet, by trial 35, the treatment group learning was significantly impaired relative to control. We used our SSRE model in a theoretical study to evaluate the design efficiency of learning experiments in terms of the number of animals per group and number of trials per animal required to characterize learning differences between two populations. Our results demonstrated that a maximum difference in the probability of a correct response between the treatment and control group learning curves of 0.07 (0.20) would require 15 to 20 (5 to 7) animals per group in an 80 (60)-trial experiment. The SSRE model offers a practical approach to dynamic analysis of population learning and a theoretical framework for optimal design of learning experiments.
Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose... more Na+ flux was studied in cultured neuroblastoma cells grown in medium containing increased glucose or L-fucose concentrations. Chronic exposure of neuroblastoma cells to 30 mM glucose or 30 mM L-fucose caused a decrease in ouabain-sensitive and veratridine-stimulated 22Na+ uptake compared with cells cultured in unsupplemented medium. The Na+ current, determined by using whole-cell configuration of the patch clamp, was also decreased in these cells. Tetrodotoxin (3 microM), which blocked whole cell Na+ currents, also blocked veratridine-stimulated 22Na+ accumulation. Culturing cells in medium containing 30 mM fructose as an osmotic control had no effect on Na+ flux. Specific [3H]saxitoxin binding was not affected by 30 mM glucose or 30 mM L-fucose compared with cells grown in unsupplemented medium, suggesting that the number of Na+ channels was not decreased. These studies suggest that exposing cultured neuronal cells to conditions that occur in the diabetic milieu alters Na+ transport and Na(+)-channel activity.
Cultured neuroblastoma, cerebral microvessel endothelial, and retinoblastoma cells were used to e... more Cultured neuroblastoma, cerebral microvessel endothelial, and retinoblastoma cells were used to examine the mechanism of acute inhibition by D-glucose of myo-inositol uptake. Acute exposure of the cells to 30 mM D-glucose caused a significant decrease in Na(+)-dependent myo-inositol uptake in all three cell types. The effect of D-glucose to acutely inhibit myo-inositol uptake was dependent on the extracellular glucose concentration and was not reversed by sorbinil. 2-Deoxy-D-glucose (30 mM), 3-O-methyl-D-glucose (30 mM), and cytochalasin B (100 microM) did not acutely inhibit myo-inositol uptake. These data suggest that the hydroxyl groups on carbons 2 and 3 of D-glucose, which in a Haworth projection appear trans to each other, are important for inhibitory activity. Other monosaccharides (30 mM) having a similar 2,3-trans-diol configuration, L-glucose, D- and L-fucose, D- and L-galactose, D- and L-xylose, and D-arabinose, all to varying degrees significantly inhibited myo-inositol uptake. In all cases, the L-isomers were more potent inhibitors of myo-inositol uptake than the corresponding D-isomers. Monosaccharides (30 mM) having hydroxyl groups on carbons 2 and 3 in a cis configuration, D-mannose, L-rhamnose, D-allose, and D-ribose, did not acutely inhibit myo-inositol uptake. Replacing the hydroxyl group with a fluorine on carbons 2 or 3 of D-glucose negated its inhibitory activity of myo-inositol uptake. In contrast, replacing the hydroxyl group with a fluorine on carbon 6 of D-glucose did not block its inhibition of myo-inositol uptake.(ABSTRACT TRUNCATED AT 250 WORDS)
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