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J M Medina

Publications and source records attributed to J M Medina.

At least 19 recordsLinked to original sources
PubMed2003

The effect of pupil size on binocular summation at suprathreshold conditions.

PURPOSE: To determine the influence of retinal illuminance on monocular or binocular visual reaction time (VRT). METHODS: On two normal subjects, uniform circular stimuli were presented with respect to a reference stimulus at the fovea under suprathreshold conditions, and the detection of positive and negative luminance variations was recorded. Binocular and monocular reaction times were measured (by the index finger pressing on a mouse key) for viewing with both natural and artificial pupils. RESULTS: Binocular reaction times were shorter than monocular reaction times; nevertheless, this binocular-summation effect was less marked in trials with the artificial pupil. Analyses of binocular-summation ratios for contrast changes for both pupil types indicated maximum and minimum binocular-saturation values depending on contrast variations in both positive and negative luminance changes. CONCLUSIONS: Binocular summation can be influenced by pupil size under suprathreshold conditions. Results are discussed in terms of retinal illuminance and cortical pupil response mechanisms.

Adult↗
PubMed2002

Binocular summation of chromatic changes as measured by visual reaction time.

We determined visual reaction times to monocular and binocular changes in the luminance of isochromatic stimuli and to monocular and binocular changes in the color of isoluminant stimuli. Two isoluminant color changes were tested: chromatic variations along the red-green axis of Boynton's (1986) two-stage color vision model and chromatic variations along the yellow-blue axis of the same model. The results indicate a greater degree of binocular summation for luminance change than for color change. This result was largely independent of the motor component of reaction time.

Adult↗
PubMed2001

Proliferation of C6 glioma cells is blunted by the increase in gap junction communication caused by tolbutamide.

We have previously reported that tolbutamide prevents the inhibition of gap junction communication in astrocytes. Here, we show that tolbutamide increases gap junction communication and connexin 43 expression in poorly coupled C6 glioma cells. The increase in communication is concurrent with the inhibition of the rate of proliferation due to a block of the progression of C6 glioma cells through the S phase of the cell cycle. The effects of tolbutamide were quantitatively similar to that found after the elevation of intracellular cAMP. Furthermore, the effects of tolbutamide and cAMP were additive. The possible beneficial effect of tolbutamide on gene therapy for gliomas is discussed.

Animals↗
PubMed2001

Oligodendrocytes use lactate as a source of energy and as a precursor of lipids.

Lactate is an important metabolic substrate for the brain during the postnatal period and also plays a crucial role in the traffic of metabolites between astrocytes and neurons. However, to date there are no clues with regard to lactate utilization by oligodendrocytes, the myelin-forming cells in the brain. In the present work, lactate utilization by oligodendrocytes in culture was investigated and compared with its utilization by cultured neurons, type 1 and type 2 astrocytes. Our results clearly indicate that oligodendrocytes readily use lactate both as a metabolic fuel and as a precursor to build carbon skeletons. Oligodendrocytes oxidize lactate at a higher rate than that observed for neurons and astrocytes, and their rate of lipid synthesis from lactate was at least 6-fold higher than that found in astrocytes or neurons. The rate of glucose utilization through different pathways was also investigated. The flux of glucose through the pentose phosphate pathway and the rate of lipid synthesis were at least 2-fold higher in oligodendrocytes than in astrocytes or neurons. These findings indicate that oligodendrocyte metabolism is designed specifically for the synthesis of lipids, presumably those of myelin.

Aging↗
PubMed2001

The enhancement of glucose uptake caused by the collapse of gap junction communication is due to an increase in astrocyte proliferation.

We have previously shown that several gap junction uncouplers increase the uptake of glucose in astrocytes. The aim of the present work was to study whether the increase in glucose uptake was a consequence of the inhibition of gap junction communication and the purpose of this effect. Our results show that alpha-glycyrrhetinic acid and endothelin-1 increase the uptake of glucose in highly, but not in poorly, coupled astrocytes. This effect depended on connexin 43 levels and was abolished when the inhibition of gap junction communication was prevented by tolbutamide or ouabain. The inhibition of gap junctions increased the rate of glucose incorporation into DNA and RNA, which was inhibited by treatment with dehydroepiandrosterone, an inhibitor of glucose-6-phosphate dehydrogenase, the regulatory enzyme of the pentose phosphate pathway. The inhibition of gap junctions significantly increased astrocyte proliferation, which was counteracted by tolbutamide. These effects were not observed in poorly coupled astrocytes expressing low levels of connexin 43. The increase in astrocyte proliferation caused by gap junction inhibition was prevented when either glucose uptake or the pentose phosphate pathway were inhibited. We conclude that the inhibition of gap junction communication induces astrocyte proliferation, resulting in an enhancement of glucose uptake and its utilization through the pentose phosphate pathway to provide ribose-5-phosphate for the synthesis of nucleic acids.

Administration, Topical↗
PubMed2001

Neuronal differentiation is triggered by oleic acid synthesized and released by astrocytes.

Unlike in the adult brain, the newborn brain specifically takes up serum albumin during the postnatal period, coinciding with the stage of maximal brain development. Here we report that albumin stimulates oleic acid synthesis by astrocytes from the main metabolic substrates available during brain development. Oleic acid released by astrocytes is used by neurons for the synthesis of phospholipids and is specifically incorporated into growth cones. Oleic acid promotes axonal growth, neuronal clustering, and expression of the axonal growth-associated protein-43, GAP-43; all these observations indicating neuronal differentiation. The effect of oleic acid on GAP-43 synthesis is brought about by the activation of protein kinase C, since it was prevented by inhibitors of this kinase, such as H-7, polymyxin or sphingosine. The expression of GAP-43 was significantly increased in neurons co-cultured with astrocytes by the presence of albumin indicating that neuronal differentiation takes place in the presence of oleic acid synthesized and released by astrocytes in situ. In conclusion, during brain development the presence of albumin could play an important role by triggering the synthesis and release of oleic acid by astrocytes, which induces neuronal differentiation.

Albumins↗
PubMed2000

ATP-sensitive potassium channel regulates astrocytic gap junction permeability by a Ca2+-independent mechanism.

Using the scrape-loading technique in cultured astrocytes, we show that sulfonylureas such as tolbutamide and glybenzcyclamide, which inhibit the ATP-sensitive K+ channel, prevent the inhibition of gap junction permeability caused by several structurally unrelated uncouplers such as oleic acid, arachidonic acid, endothelin-1, octanol, and alpha-glycyrrhetinic acid. When the intracellular level of Ca2+ was diminished, all the uncouplers tested were still able to inhibit gap junction communication, indicating that their inhibitory effect was not mediated by Ca2+. In addition, tolbutamide and glybenzcyclamide prevented the inhibitory effect of these uncouplers in Ca(2+)-depleted astrocytes, suggesting that the inhibition of the ATP-sensitive K+ channel increases gap junction permeability through a Ca(2+)-independent mechanism. The activation of the ATP-sensitive K+ channel caused by potassium channel openers such as diazoxide and pinacidil led to the inhibition of gap junction communication and overcame the effect of sulfonylureas. These results suggest that the ATP-sensitive K+ channel regulates gap junctional permeability.

Adenosine Triphosphate↗
PubMed1999

Nitric oxide mediates brain mitochondrial maturation immediately after birth.

The possible role of nitric oxide (*NO) in brain mitochondrial maturation was studied. Within the first 5 min after birth, a sharp increase in ATP concentrations was observed, coinciding with an increase in mitochondrial complex II-III (succinate-cytochrome c reductase) activity, while complex I (NADH-CoQ1 reductase) and complex IV (cytochrome c oxidase) activities remained unchanged. Under the same circumstances, cGMP concentrations were increased by 5 min after birth, correlating significantly with ATP concentrations. Since ATP concentrations also correlated significantly with mitochondrial complex II-III activity, these three parameters may be associated. Inhibition of *NO synthase activity brought about by the administration of N(omega)-nitro-L-arginine monomethyl ester to mothers prevented the postnatal increase in cGMP and ATP levels and complex II-III activity. These results suggest that early postnatal mitochondrial maturation in the brain is a *NO-mediated process.

Adenosine Triphosphate↗
PubMed1999

Nitric oxide mediates glutamate-induced mitochondrial depolarization in rat cortical neurons.

Mitochondria have been considered to be a target for glutamate neurotoxicity. The aim of the present work was to investigate the mechanisms leading to glutamate-mediated mitochondrial deenergization, as measured by mitochondrial membrane potential and cell respiration in cultured neurons. Glutamate exposure to cells induced pronounced mitochondrial depolarization associated with an impairment in neuronal respiration, leading to neuronal ATP depletion. These effects were prevented by both the nitric oxide (. NO) synthase inhibitor Nomega-nitro-l-arginine methyl ester and by the N-methyl-d-aspartate glutamate-subtype receptor inhibitor d-(-)-2-amino-5-phosphopentanoate. Our results suggest that glutamate causes ATP depletion by collapsing mitochondrial membrane potential through a.NO-mediated mechanism.

Animals↗
PubMed1999

Peroxynitrite anion stimulates arginine release from cultured rat astrocytes.

The biosynthesis of the physiological messenger nitric oxide (*NO) in neuronal cells is thought to depend on a glial-derived supply of the *NO synthase substrate arginine. To expand our knowledge of the mechanism responsible for this glial-neuronal interaction, we studied the possible roles of peroxynitrite anion (ONOO-), superoxide anion (O2*-), *NO, and H2O2 in L-[3H]arginine release in cultured rat astrocytes. After 5 min of incubation at 37 degrees C, initial concentrations of 0.05-2 mM ONOO- stimulated the release of arginine from astrocytes in a concentration-dependent way; this effect was maximum from 1 mM ONOO- and proved to be approximately 400% as compared with control cells. ONOO(-)-mediated arginine release was prevented by arginine transport inhibitors, such as L-lysine and N(G)-monomethyl-L-arginine, suggesting an involvement of the arginine transporter in the effect of ONOO-. In situ xanthine/xanthine oxidase-generated O2*- (20 nmol/min) stimulated arginine release to a similar extent to that found with 0.1 mM ONOO-, but this effect was not prevented by arginine transport inhibitors. *NO donors, such as sodium nitroprusside, S-nitroso-N-acetylpenicillamine, or 1-[2-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium+ ++-1,2-diolate, and H2O2 did not significantly modify arginine release. As limited arginine availability for neuronal *NO synthase activity may be neurotoxic due to ONOO- formation, our results suggest that ONOO(-)-mediated arginine release from astrocytes may contribute to replenishing neuronal arginine, hence avoiding further generation of ONOO- within these cells.

Animals↗
PubMed1999

Induction of glucose-6-phosphate dehydrogenase by lipopolysaccharide contributes to preventing nitric oxide-mediated glutathione depletion in cultured rat astrocytes.

Treatment of cultured rat astrocytes with lipopolysaccharide (LPS; 1 microg/ml) increased mRNA expression of glucose-6-phosphate dehydrogenase (G6PD), the rate-limiting step in the pentose phosphate pathway (PPP), in a time-dependent fashion (0-24 h). This effect was accompanied by an increase in G6PD activity (1.74-fold) and in the rate of glucose oxidation through the PPP (6.32-fold). Inhibition of inducible nitric oxide synthase (iNOS) activity by 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (AMT; 50 microM) did not alter the LPS-mediated enhancement of G6PD mRNA expression or PPP activity. Blockade of nuclear factor-kappaB (NF-kappaB) activation by N-benzyloxycarbonyl-Ile-Glu-(O-tert-butyl)-Ala-leucinal (1 microM) prevented the expression of both iNOS mRNA and G6PD mRNA, suggesting that iNOS and G6PD are co-induced by LPS through a common transcriptional pathway involving NF-kappaB activation. Incubation of cells with LPS for 24 h increased intracellular NADPH concentrations (1.63-fold) as compared with untreated cells, but GSH concentrations were not modified by LPS treatment up to 60 h of incubation. However, inhibition of G6PD activity by dehydroepiandrosterone (DHEA; 100 microM), which prevented LPS-mediated enhancements in PPP activity and NADPH concentrations, caused a 50% decrease in the GSH/GSSG ratio after 24-36 h and in GSH concentrations after 60 h of incubation. Furthermore, the changes in glutathione concentrations caused by DHEA were abolished by AMT, suggesting that nitric oxide and/or its reactive derivatives would be involved in this process. From these results, we conclude that LPS-mediated G6PD expression prevents GSH depletion due to nitric oxide and suggest that this phenomenon may be a contributing factor in the defense mechanisms that protect astrocytes against nitric oxide-mediated cell injury.

Adjuvants, Immunologic↗
PubMed1999

The effect of albumin on astrocyte energy metabolism is not brought about through the control of cytosolic Ca2+ concentrations but by free-fatty acid sequestration.

Albumin is an important serum protein that under normal circumstances is not present in the brain. However, during development, under hypoxia, or after breakdown of the blood-brain barrier, albumin is found in the brain, where it is able to regulate energy metabolism. In this work the mechanism through which albumin regulates astrocyte metabolism was investigated. Our results show that albumin strongly increases (more than 100%) the flux of glucose and lactate through the pyruvate dehydrogenase-catalyzed reaction in astrocytes from primary culture. However, albumin only slightly stimulated other metabolic pathways, such as the tricarboxylic acid cycle or the pentose phosphate shunt, indicating that it exerts its effect specifically on the reaction catalyzed by pyruvate dehydrogenase. Although albumin increased cytosolic Ca2+ concentrations in astrocytes, our results show that the increase in pyruvate dehydrogenase activity promoted by albumin is not due to the enhancement of Ca2+ concentrations. Indeed, highly purified albumins failed to increase the Ca2+ concentration but did enhance lactate oxidation. In agreement with this, the effect of albumin on lactate oxidation was not abolished after Ca2+ depletion. Instead, the presence of fatty acids inhibited lactate oxidation and counteracted the effect of albumin, suggesting that albumin activates pyruvate dehydrogenase by binding free fatty acids and/or their CoA-derivatives.

Albumins↗
PubMed1998

The K-ATP channel regulates the effect of Ca2+ on gap junction permeability in cultured astrocytes.

Using the scrape-loading technique we show that tolbutamide and glybenzcyclamide, two inhibitors of the K+ channel sensitive to ATP (K-ATP channel), partially prevent the inhibition of gap junction permeability promoted by Ca2+ in cultured astrocytes. This effect was dose-dependent, reaching a maximum at 400 microM and 1 microM of tolbutamide and glybenzcyclamide, respectively. The presence of the Ca2+ ionophore A-23187 strongly reduced the concentration of Ca2+ required to block gap junction permeability but did not abolish the effect of tolbutamide and glybenzcyclamide. These results suggest that the effect of these two compounds are not brought about by control of the intracellular concentration of Ca2+ but probably by the promotion of plasma membrane depolarization.

ATP-Binding Cassette Transporters↗
PubMed1998

Glutamate neurotoxicity is associated with nitric oxide-mediated mitochondrial dysfunction and glutathione depletion.

The role of mitochondrial energy metabolism in glutamate mediated neurotoxicity was studied in rat neurones in primary culture. A brief (15 min) exposure of the neurones to glutamate caused a dose-dependent (0.01-1 mM) increase in cyclic GMP levels together with delayed (24 h) neurotoxicity and ATP depletion. These effects were prevented by either the nitric oxide (.NO) synthase (NOS) inhibitor Nomega-nitro-L-arginine methyl ester (NAME; 1 mM) or by the N-methyl-D-aspartate (NMDA) glutamate-subtype receptor antagonist D-(-)-2-amino-5-phosphonopentanoate (APV; 0.1 mM). Glutamate exposure (0.1 mM and 1 mM) followed by 24 h of incubation caused the inhibition of succinate-cytochrome c reductase (20-25%) and cytochrome c oxidase (31%) activities in the surviving neurones, without affecting NADH-coenzyme-Q1 reductase activity. The rate of oxygen consumption was impaired in neurones exposed to 1 mM glutamate, either with glucose (by 26%) or succinate (by 39%) as substrates. These effects on the mitochondrial respiratory chain and neuronal respiration, together with the observed glutathione depletion (20%) by glutamate exposure were completely prevented by NAME or APV. Our results suggest that mitochondrial dysfunction and impairment of antioxidant status may account for glutamate-mediated neurotoxicity via a mechanism involving .NO biosynthesis.

2-Amino-5-phosphonovalerate↗
PubMed1998

Nitric oxide mediates brain mitochondrial damage during perinatal anoxia.

The possible role of nitric oxide (.NO) in brain energy metabolism during perinatal asphyxia in the rat was studied. Exposure of early neonates to 5 min of anoxia significantly inhibited brain mitochondrial complex II-III activity by 25%, without affecting complex I, complex IV or citrate synthase activities. This insult was accompanied by ATP depletion (54%) and increased concentration of nitrites plus nitrates (1.4-fold), suggesting enhanced .NO synthesis. Administration of Nomega-nitro-L-arginine monomethyl ester (L-NAME) to the mothers inhibited neonatal brain .NO synthase activity, as reflected by the decreased (23%) cyclic GMP concentration. These L-NAME-treated neonates showed complete resistance to anoxic-mediated brain mitochondrial complex II-III damage. Our results suggest that brain mitochondrial dysfunction leading to energy deficiency during perinatal asphyxia is a .NO-mediated process.

Animals↗
PubMed1998

A rapid method for the isolation of metabolically active mitochondria from rat neurons and astrocytes in primary culture.

A rapid method (about 1.5 h) for the isolation of intact functional mitochondria from neurons and astrocytes in primary culture is described. Mitochondria isolated by this method are metabolically active and tightly coupled as shown by respiratory control ratio values, which were about 4 with glutamate-malate as substrate. The activities of marker enzymes revealed the occurrence of a low degree of cytosolic (5%) or synaptosomal (5.5%) contamination in the mitochondrial fractions. In addition, the activity of citrate synthase was increased by 4 fold in both neuronal and astrocytic mitochondria with respect to values found in cell homogenates. These results confirm that the method affords mitochondrial preparations from cultured brain cells at suitable levels of purity and enrichment for the study of their mitochondrial function. Since mitochondrial damage has been associated with the pathogenesis of certain neurodegenerative diseases, such as Alzheimer's and Parkinson's diseases (P. Chagnon, C. Betard, Y. Robitaille, A. Cholette, D. Gauvreau, Distribution of brain cytochrome oxidase activity in various neurodegenerative disease, Neuroreport 6 (1995) 711-715 [6]; S.J. Kish, C. Bergeron, A. Rajput, S. Dozic, F. Mastrogiacomo, L. Chang, J.M. Wilson, L.M. DiStefano, J.N. Nobrega, Brain cytochrome oxidase in Alzheimer's disease, J. Neurochem. 59 (1992) 776-779 [10]; A.H.V. Schapira, J.M. Cooper, D. Dexter, J.B. Clark, P. Jenner, C.D. Marsden, Mitochondrial complex I deficiency in Parkinson's disease, J. Neurochem. 54 (1990) 823-827 [15]), the method described here shed light on the possible susceptibility of neuronal or astrocytic mitochondria to deleterious effects of these diseases.

Animals↗
PubMed1997

cAMP and Ca2+ involvement in the mitochondrial response of cultured fetal rat hepatocytes to adrenaline.

The effect of adrenaline on the control of respiratory activity of mitochondria from fetal hepatocytes in primary culture was studied. In the absence of adrenaline, the respiratory control ratio (RCR) of mitochondria increased during the first 3 days of culture due to a decrease in the rate of state 4 respiration. The presence of adrenaline in the incubation medium further increased the mitochondrial RCR through a decrease in the rate of respiration in state 4 and to an increase in the respiration rate in state 3. The effect of adrenaline was mimicked by dibutyryl-cAMP, forskolin, and isobutyl methyl xanthine. All these compounds increased cAMP concentrations, suggesting that cAMP may be involved in the effect of adrenaline. The increase in intracellular free Ca2+ concentrations caused by phenylephrine, vasopressin, or thapsigargin was also accompanied by an increase in the RCR, suggesting that both phenomena are associated. Dibutyryl-cAMP also increased free Ca2+ concentrations, suggesting that the effects of cAMP may be mediated by free Ca2+ concentrations. Adrenaline, dibutyryl-cAMP, phenylephrine, vasopressin, and thapsigargin promoted adenine nucleotide accumulation in mitochondria; this may be an intermediate step in the activation of mitochondrial respiratory function. These results suggest that the stimulatory effect of adrenaline on mitochondrial maturation in cultured fetal rat hepatocytes may be exerted through a mechanism in which both cAMP and Ca2+ act as second messengers. It is concluded that the effect of adrenaline on mitochondrial maturation is exerted by both alpha- and beta-adrenergic mechanisms and is mediated by the increase in adenine nucleotide contents of mitochondria.

Adenine Nucleotides↗