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ABSTRACT: The marine alga Ulva compressa exposed to 10 µM copper showed a triphasic increase of intracellular calcium with maximal levels at 2, 3 and 12 h involving the activation of ryanodine-, Ins(1,4,5)P 3- and NAADP-sensitive calcium channels. In order to analyze the requirement of extracellular calcium entry for intracellular calcium release as well as the activation of voltage-dependent calcium channels (VDCC) and phospholipase C, U. compressa was treated with EGTA, a non-permeable calcium chelating agent, with verapamil, nipfedipine and diltiazem, inhibitors of L-type VDCC, and with neomycin and U731222, inhibitors of phospholipase C. The release of intracellular calcium was partially inhibited with EGTA at 2 and 3 h and completely inhibited at 12 h of copper exposure and decreased with inhibitors of L-type VDCC and phospholipase C. Thus, copper-induced intracellular calcium release depends on calcium entry and activation of L-type VDCC and phospholipase C. An integrative model of copper-induced cellular responses in U. compressa is presented.
Plant signaling & behavior 07/2012; 7(7):728-32.
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ABSTRACT: To analyze the copper-induced cross talk among calcium, nitric oxide (NO), and hydrogen peroxide (H(2)O(2)) and the calcium-dependent activation of gene expression, the marine alga Ulva compressa was treated with the inhibitors of calcium channels, ned-19, ryanodine, and xestospongin C, of chloroplasts and mitochondrial electron transport chains, 3-(3,4-dichlorophenyl)-1,1-dimethylurea and antimycin A, of pyruvate dehydrogenase, moniliformin, of calmodulins, N-(6-aminohexyl)-5-chloro-1-naphtalene sulfonamide, and of calcium-dependent protein kinases, staurosporine, as well as with the scavengers of NO, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, and of H(2)O(2), ascorbate, and exposed to a sublethal concentration of copper (10 μm) for 24 h. The level of NO increased at 2 and 12 h. The first peak was inhibited by ned-19 and 3-(2,3-dichlorophenyl)-1,1-dimethylurea and the second peak by ned-19 and antimycin A, indicating that NO synthesis is dependent on calcium release and occurs in organelles. The level of H(2)O(2) increased at 2, 3, and 12 h and was inhibited by ned-19, ryanodine, xestospongin C, and moniliformin, indicating that H(2)O(2) accumulation is dependent on calcium release and Krebs cycle activity. In addition, pyruvate dehydrogenase, 2-oxoxglutarate dehydrogenase, and isocitrate dehydrogenase activities of the Krebs cycle increased at 2, 3, 12, and/or 14 h, and these increases were inhibited in vitro by EGTA, a calcium chelating agent. Calcium release at 2, 3, and 12 h was inhibited by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide and ascorbate, indicating activation by NO and H(2)O(2). In addition, the level of antioxidant protein gene transcripts decreased with N-(6-aminohexyl)-5-chloro-1-naphtalene sulfonamide and staurosporine. Thus, there is a copper-induced cross talk among calcium, H(2)O(2), and NO and a calcium-dependent activation of gene expression involving calmodulins and calcium-dependent protein kinases.
Plant physiology 03/2012; 158(3):1451-62. · 6.53 Impact Factor
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ABSTRACT: Zn²(+) is an essential ion that is stored in and co-released from glutamatergic synapses and it modulates neurotransmitter receptors involved in long-term potentiation (LTP). However, the mechanism(s) underlying Zn²(+) -induced modulation of LTP remain(s) unclear. As the purinergic P2X receptors are relevant targets for Zn²(+) action, we have studied their role in LTP modulation by Zn²(+) in the CA1 region of rat hippocampal slices. Induction of LTP in the presence of Zn²(+) revealed a biphasic effect - 5-50 μm enhanced LTP induction, whereas 100-300 μm Zn²(+) inhibited LTP. The involvement of a purinergic mechanism is supported by the fact that application of the P2X receptor antagonists 2',3'-O-(2,4,6-trinitrophenyl) ATP (TNP-ATP) and periodate-oxidized ATP fully abolished the facilitatory effect of Zn²(+) . Notably, application of the P2X₇ receptor-specific antagonist Brilliant Blue G did not modify the Zn²(+) -dependent facilitation of LTP. Exogenous ATP also produced a biphasic effect - 0.1-1 μm ATP facilitated LTP, whereas 5-10 μm inhibited LTP. The facilitatory effect of ATP was abolished by the application of TNP-ATP and was modified in the presence of 5 μm Zn²(+) , suggesting that P2X receptors are involved in LTP induction and that Zn²(+) leads to an increase in the affinity of P2X receptors for ATP. The latter confirms our previous results from heterologous expression systems. Collectively, our results indicate that Zn²(+) at low concentrations enhances LTP by modulating P2X receptors. Although it is not yet clear which purinergic receptor subtype(s) is responsible for these effects on LTP, the data presented here suggest that P2X₄ but not P2X₇ is involved.
European Journal of Neuroscience 02/2011; 33(7):1175-85. · 3.63 Impact Factor
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ABSTRACT: The marine alga Ulva compressa (Chlorophyta) showed a triphasic release of intracellular calcium with maximal levels at 2, 3 and 12 h and a biphasic accumulation of intracellular hydrogen peroxide with peaks at 3 and 12 h when cultivated with copper excess. Intracellular hydrogen peroxide originated exclusively in organelles. In this work, we analyzed the intracellular origin of calcium release and the type of calcium channels activated in response to copper excess. U. compressa was treated with thapsigargin, an inhibitor of endoplasmic reticulum (ER) calcium ATPase, ryanodine, an inhibitor of ryanodine-sensitive channels, and xestospongin C, an inhibitor of inositol 1, 4, 5-triphosphate (IP(3))-sensitive channels. Thapsigargin induced the depletion of calcium stored in ER at 75 min and completely inhibited calcium release at 2, 3 and 12 h of copper exposure indicating that calcium release originated in ER. In addition, ryanodine and xestospogin C inhibited calcium release at 2 and 3 h of copper exposure whereas the peak at 12 h was only inhibited by ryanodine. Thus, copper induced the activation of ryanodine-sensitive and IP(3)-sensitive calcium channels in ER of U. compressa.
Plant signaling & behavior 12/2010; 5(12):1647-9.
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ABSTRACT: In order to analyse copper-induced calcium release and (reactive oxygen species) ROS accumulation and their role in antioxidant and defense enzymes activation, the marine alga Ulva compressa was exposed to 10 µM copper for 7 d. The level of calcium, extracellular hydrogen peroxide (eHP), intracellular hydrogen peroxide (iHP) and superoxide anions (SA) as well as the activities of ascorbate peroxidase (AP), glutathione reductase (GR), glutathione-S-transferase (GST), phenylalanine ammonia lyase (PAL) and lipoxygenase (LOX) were determined. Calcium release showed a triphasic pattern with peaks at 2, 3 and 12 h. The second peak was coincident with increases in eHP and iHP and the third peak with the second increase of iHP. A delayed wave of SA occurred after day 3 and was not accompanied by calcium release. The accumulation of iHP and SA was mainly inhibited by organellar electron transport chains inhibitors (OETCI), whereas calcium release was inhibited by ryanodine. AP activation ceased almost completely after the use of OETCI. On the other hand, GR and GST activities were partially inhibited, whereas defense enzymes were not inhibited. In contrast, PAL and LOX were inhibited by ryanodine, whereas AP was not inhibited. Thus, copper stress induces calcium release and organellar ROS accumulation that determine the differential activation of antioxidant and defense enzymes.
Plant Cell and Environment 04/2010; 33(10):1627-40. · 5.22 Impact Factor
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ABSTRACT: 3,4-Methylenedioxymethamphetamine (MDMA), an important recreational psychostimulant drug, was examined for its ability to alter visuo-spatial learning and synaptic plasticity. Young rats received MDMA (0.2 and 2mg/kg s.c.) twice per day for 6 days while their visuo-spatial learning was tested using the Morris Water Maze. After this, animals were sacrificed and LTP induced in hippocampal slices. Visuo-spatial learning was impaired and LTP reduced, both dose-dependently, without changes in serotonin levels or paired-pulse facilitation. We conclude that low, nontoxic doses of MDMA, applied during several days, slow learning by impairing postsynaptic plasticity.
Neuroscience Letters 12/2009; 469(3):375-9. · 2.11 Impact Factor
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ABSTRACT: The epithelial-mesenchymal transition (EMT) is considered a key step in tumor progression, where the invasive cancer cells change from epithelial to mesenchymal phenotype. During this process, a decrease or loss in adhesion molecules expression and an increase in migration molecules expression are observed. The aim of this work was to determine the expression and cellular distribution of syndecan-1 and -2 (migration molecules) and E-cadherin and beta-catenin (adhesion molecules) in different stages of prostate cancer progression. A quantitative immunohistochemical study of these molecules was carried out in tissue samples from benign prostatic hyperplasia and prostate carcinoma, with low and high Gleason score, obtained from biopsies archives of the Clinic Hospital of the University of Chile and Dipreca Hospital. Polyclonal specific antibodies and amplification system of estreptavidin-biotin peroxidase and diaminobenzidine were used. Syndecan-1 was uniformly expressed in basolateral membranes of normal epithelium, changing to a granular cytoplasmatic expression pattern in carcinomas. Syndecan-2 was observed mainly in a cytoplasmatic granular pattern, with high immunostaining intensity in areas of low Gleason score. E-cadherin was detected in basolateral membrane of normal epithelia showing decreased expression in high Gleason score samples. beta-Catenin was found in cell membranes of normal epithelia changing its distribution toward the nucleus and cytoplasm in carcinoma samples. We concluded that changes in expression and cell distribution of E-cadherin and beta-catenin correlated with the progression degree of prostate adenocarcinoma, suggesting a role of these molecules as markers of progression and prognosis. Furthermore, changes in the pattern expression of syndecan-1 and -2 indicate that both molecules may be involved in the EMT and tumor progression of prostate cancer.
Urologic Oncology 06/2009; 28(5):534-40. · 3.22 Impact Factor
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ABSTRACT: Dichlorvos is the active molecule of the pro-drug metrifonate used to revert the cognitive deficits associated with Alzheimer's disease. A few years ago it was reported that dichlorvos inhibits the enzyme acylpeptide hydrolase at lower doses than those necessary to inhibit acetylcholinesterase to the same extent. Therefore, the aim of our investigation was to test the hypothesis that dichlorvos can enhance synaptic efficacy through a mechanism that involves acylpeptide hydrolase instead of acetylcholinesterase inhibition. We used long-term potentiation induced in rat hippocampal slices as a model of synaptic plasticity. Our results indicate that short-term exposures (20 min) to 50 microM dichlorvos enhance long-term potentiation in about 200% compared to the control condition. This effect is correlated with approximately 60% inhibition of acylpeptide hydrolase activity, whereas acetylcholinesterase activity remains unaffected. Paired-pulse facilitation and inhibition experiments indicate that dichlorvos does not have any presynaptic effect in the CA3-->CA1 pathway nor affect gabaergic interneurons. Interestingly, the application of 100 nM methyllicaconitine, an alpha(7) nicotinic receptor antagonist, blocked the enhancing effect of dichlorvos on long-term potentiation. These results indicate that under the exposure conditions described above, dichlorvos enhances long-term potentiation through a postsynaptic mechanism that involves (a) the inhibition of the enzyme acylpeptide hydrolase and (b) the modulation of alpha(7) nicotinic receptors.
Toxicology and Applied Pharmacology 05/2009; 238(1):37-46. · 4.45 Impact Factor
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Verónica Donoso,
Christian R Gomez,
Miguel Angel Orriantia,
Viviana Pérez,
Claudio Torres,
Claudio Coddou,
Pablo Nelson,
Kevin Maisey, Bernardo Morales,
Ricardo Fernandez,
Mónica Imarai,
Juan Pablo Huidobro-Toro,
Felipe Sierra,
Claudio Acuña-Castillo
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ABSTRACT: Aging results in a general decline in the response to external insults, including acute inflammatory challenges. In young animals, the inflammatory response requires activation of the sympathetic system, including neurotransmitters such as ATP, and catecholamines (epinephrine and norepinephrine). To test whether aging affects activation of this axis, and whether this in turn might affect cytokine release, we administered lipopolysaccharide (LPS) i.p. to adult, middle-aged and aged Fisher 344 rats (6-, 15- and 23-month old, respectively) and evaluated the early (0-12h) serum levels of Neuropeptide-Y (NP-Y), ATP and vanillyl mandelic acid (VMA, as an indirect measurement of catecholamine levels). In addition, we evaluated the association between these factors and serum levels of the cytokines tumor necrosis factor-alpha (TNFalpha) and interleukin-10 (IL-10). Induction of both ATP and NP-Y was markedly reduced in the serum of aged animals, when compared to their younger counterparts, while induction of VMA was not affected by age. In spite of these changes, serum levels of TNFalpha and IL-10 were strongly hyper induced and delayed in aged rats. The results suggest that during aging there is a dysregulation in sympathetic neurotransmitter regulatory mechanisms, and this might play a role in the impairment of the inflammatory response.
Mechanisms of Ageing and Development 11/2008; 129(12):728-34. · 3.44 Impact Factor
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ABSTRACT: We have demonstrated that adenosine did not produce any change of intracellular free Ca2+ concentration ([Ca2+]i) in oviductal ciliated cells; however, it increased the ATP-induced Ca2+ influx through the activation of protein kinase A (PKA). Uncaging of IP3 and cAMP triggered a larger Ca2+ influx than did IP3 alone. Furthermore, the IP3 effect was abolished by Xestospongin C, an IP3 receptor blocker. Whole-cell recordings demonstrated the presence of an ATP-induced Ca2+ current, and the addition of adenosine increased the peak of this current. This effect was not observed in the presence of H-89, a PKA inhibitor. Using excised macro-patches of plasma membrane, IP3 generated a current, which was higher in the presence of the catalytic PKA subunit and this current was blocked by Xestospongin C. We show here that activation of plasma membrane IP3 receptors directly triggers Ca2+ influx in response to ATP and that these receptors are modulated by adenosine-activated PKA.
Biochemical and Biophysical Research Communications 01/2008; 364(4):815-21. · 2.48 Impact Factor
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ABSTRACT: Organophosphate pesticides have been classically described as inhibitors of acetylcholinesterase (AChE) activity in insects and invertebrates. However, there is now more evidence supporting the hypothesis that these compounds also act through noncholinergic pathways, especially those related to cognitive processes. The enzyme acylpeptide hydrolase was identified as a new target for organophosphate pesticides. This enzyme is more sensitive than AChE to some organophosphates (OP), including dichlorvos, which is the parent compound for metrifonate, a therapeutic agent used in the treatment of cognitive impairment associated to Alzheimer's disease. Therefore, there is some doubt as to whether the mechanism of action of this drug is mediated by a potentiation of cholinergic transmission. However, the direct action of acylpeptide hydrolase in cognitive processes and the physiological and molecular mechanisms underlying subacute exposure to OP have yet to be demonstrated. This review deals with evidence demonstrating the existence of mechanisms of actions of OP, which are independent of cholinergic pathway potentiation and which have an effect on cognitive processes. In addition, the possible participation of the enzyme acylpeptide hydrolase in these processes is also discussed. Finally, the possibility of using this enzyme activity as a new biomarker for exposure to OP is considered.
Journal of Toxicology and Environmental Health Part B 01/2008; 10(8):623-30. · 4.72 Impact Factor
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ABSTRACT: Despite its still rapidly increasing use in the treatment of the attention deficit/hyperactivity disorder, the effects of methylphenidate on behavior and learning are not yet fully understood. We have used the Morris water maze to study effects of methylphenidate (1 mg/kg) on target-oriented behavior and visuospatial learning in young rats. Although the relatively low dose of 1 mg/kg methylphenidate changed the behavior in the Morris water maze when the goal was visible, reducing times to reach the visible platform, learning of the position of the hidden platform, was not influenced, nor was 'relearning' after shifting the position of the platform. These results indicate that methylphenidate can influence goal-oriented behavior at doses that do not change visuospatial learning.
Neuroreport 08/2007; 18(10):1059-62. · 1.66 Impact Factor
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ABSTRACT: During a brief postnatal critical period, excitatory connections in visual cortex can be easily modified by alterations of visual experience. Recent studies conducted in rodents, and particularly in genetically altered mice, have implicated the maturation of cortical GABAergic inhibition in the timing of the critical period. In this paper we (1) review the postnatal changes in GABAergic transmission that can have consequences for visual cortex plasticity and (2) discuss possible mechanisms by which GABAergic circuits could regulate the onset and termination of the critical period for cortical plasticity.
Brain Research Reviews 01/2006; 50(1):126-33. · 10.34 Impact Factor
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ABSTRACT: Cells can be considered as integrators of simultaneous stimuli, in which cross-talk between transduction pathways can eventually produce responses that are significantly different from simply additive responses. Synergism represents an efficient means of increasing the amplitude of cellular responses induced by low levels of stimulation. Recently, several kinetic and physicochemical models have been developed to describe and predict synergistic responses. In this article, the mechanisms that control the magnitude and timing of cellular synergism are discussed. We suggest that the analysis of theoretical models could enable a general prediction of synergism despite the presence of signal-specific synergistic responses. In addition, application of the proposed concepts should aid understanding of the wide occurrence of synergism induced by interacting transduction pathways in multi-drug clinical treatment.
Trends in Pharmacological Sciences 11/2005; 26(10):526-32. · 10.93 Impact Factor
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ABSTRACT: The volume of oviductal fluid fluctuates during the estrous cycle, suggesting that water availability is under hormonal control. It has been postulated that sex-steroid hormones may regulate aquaporin (AQP) channels involved in water movement across cell membranes. Using a functional assay (oocytes of Xenopus laevis), we demonstrated that the rat oviductal epithelium contains mRNAs coding for water channels, and we identified by RT-PCR the mRNAs for AQP5, -8, and -9, but not for AQP2 and -3. The immunoreactivity for AQP5, -8, and -9 was localized only in epithelial cells of the oviduct. The distribution of AQP5 and -8 was mainly cytoplasmic, whereas we confirmed, by confocal microscopy, that AQP9 localized to the apical plasma membrane. Staining of AQP5, -8, and -9 was lost after ovariectomy, and only AQP9 immunoreactivity was restored after estradiol and/or progesterone treatments. The recovery of AQP9 reactivity after ovariectomy correlated with increased mRNA and protein levels after treatment with estradiol alone or progesterone administration after estradiol priming. Interestingly, progesterone administration after progesterone priming also induced AQP9 expression but without a change in mRNA levels. Levels of AQP9 varied along the estrous cycle with their highest levels during proestrus and estrus. These results indicate that steroid hormones regulate AQP9 expression at the mRNA and protein level and that other ovarian signals are involved in the expression of AQP5 and -8. Thus hormonal regulation of the type and quantity of water channels in this epithelium might control water transport in the oviductal lumen.
AJP Cell Physiology 06/2005; 288(5):C1048-57. · 3.54 Impact Factor
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ABSTRACT: An increase in intracellular free Ca(2+) concentration ([Ca(2+)](i)) has been shown to be involved in the increase in ciliary beat frequency (CBF) in response to ATP; however, the signaling pathways associated with inositol 1,4,5-trisphosphate (IP(3)) receptor-dependent Ca(2+) mobilization remain unresolved. Using radioimmunoassay techniques, we have demonstrated the appearance of two IP(3) peaks occurring 10 and 60 s after ATP addition, which was strongly correlated with a release of intracellular Ca(2+) from internal stores and an influx of extracellular Ca(2+), respectively. In addition, ATP-dependent Ca(2+) mobilization required protein kinase C (PKC) and Ca(2+)/calmodulin-dependent protein kinase II activation. We found an increase in PKC activity in response to ATP, with a peak at 60 s after ATP addition. Xestospongin C, an IP(3) receptor blocker, significantly diminished both the ATP-induced increase in CBF and the initial transient [Ca(2+)](i) component. ATP addition in the presence of xestospongin C or thapsigargin revealed that the Ca(2+) influx is also dependent on IP(3) receptor activation. Immunofluorescence and confocal microscopic studies showed the presence of IP(3) receptor types 1 and 3 in cultured ciliated cells. Immunogold electron microscopy localized IP(3) receptor type 3 to the nucleus, the endoplasmic reticulum, and, interestingly, the plasma membrane. In contrast, IP(3) receptor type 1 was found exclusively in the nucleus and the endoplasmic reticulum. Our study demonstrates for the first time the presence of IP(3) receptor type 3 in the plasma membrane in ciliated cells and leads us to postulate that the IP(3) receptor can directly trigger Ca(2+) influx in response to ATP.
AJP Cell Physiology 11/2004; 287(4):C1114-24. · 3.54 Impact Factor
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ABSTRACT: S-methylcysteine (SMC) is formed after exposure to monohalomethanes in rodents as well as in humans. The present study was performed to study whether SMC, directly or indirectly, contributes to the well-known neurotoxicity of monohalomethanes. We have investigated the effects of acute exposure to SMC by means of electrophysiolocal measurements in freshly prepared hippocampal slices and dissociated hippocampal neurons in culture. For longer-term exposures (24 h) we have used organotypic cultures (2 weeks in culture), taking electrophysiologic recordings and assessing membrane integrity with propidium iodide (PI) fluorescence. We found that only high concentrations of SMC (10(-2) M; exposure time 30 min) in freshly isolated slices of adult rats reduce synaptically evoked population spikes in the CA1 region. This effect was at least partially reversible. In organotypic cultures, at 5 x 10(-5) M after 24 h of exposure, SMC compromises membrane integrity as revealed by PI fluorescence, only in the dentate gyrus, spreading to pyramidal cell layers at 50 x 10(-4) M. At 5 x 10(-6) and 2 x 10(-5) M, under the same experimental conditions, no changes were seen with the PI method, but we recorded increased population spike amplitudes, repetitive discharges and frequency potentiation (at a stimulus repetition rate of 0.05 Hz). Using whole-cell patch clamp in hippocampal dissociated neurons we have found that SMC (applied for approximately 1s) reduces GABA-induced currents ( IC(50) = 4.4 x 10(-4) M) without having an effect of its own, acting like a competitive antagonist at GABA(A) receptors. Our findings are in line with the view that the ability of monohalomethanes to induce the formation of SMC is an important factor for their neurotoxicity, provided that SMC is allowed to act at least for several hours. The effects exerted by SMC seem to be due, at least in part, to its interaction with GABA receptors.
NeuroToxicology 10/2004; 25(5):817-23. · 3.10 Impact Factor
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ABSTRACT: Visual deprivation such as dark rearing (DR) prolongs the critical period for ocular dominance plasticity and retards the maturation of gamma-aminobutyric acid (GABA)ergic inhibition in visual cortex. The molecular signals that mediate the effects of DR on the development of visual cortex are not well defined. To test the role of brain-derived neurotrophic factor (BDNF), we examined the effects of DR in transgenic mice in which BDNF expression in visual cortex was uncoupled from visual experience and remained elevated during DR. In dark-reared transgenic mice, visual acuity, receptive field size of visual cortical neurons, critical period for ocular dominance plasticity, and intracortical inhibition were indistinguishable from those observed in light-reared mice. Therefore, BDNF overexpression is sufficient for the development of aspects of visual cortex in the absence of visual experience. These results suggest that reduced BDNF expression contributes to retarded maturation of GABAergic inhibition and delayed development of visual cortex during visual deprivation.
Proceedings of the National Academy of Sciences 11/2003; 100(21):12486-91. · 9.68 Impact Factor
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ABSTRACT: To elucidate the role of extracellular histidines in the modulation of the rat P2X4 receptor by trace metals, we generated single, double, and triple histidine mutants for residues 140, 241, and 286, replacing them with alanines. cDNAs for the wild-type and receptor mutants were expressed in Xenopus laevis oocytes and in human embryonic kidney 293 cells and examined by the two electrode and patch clamp techniques, respectively. Whereas copper inhibited concentration-dependently the ATP-gated currents in the wild-type and in the single or double H241A and H286A receptor mutants, all receptors containing H140A were insensitive to copper in both cell systems. The characteristic bell-shaped concentration-response curve of zinc observed in the wild-type receptor became sigmoid in both oocytes and human embryonic kidney cells expressing the H140A mutant; in these mutants, the zinc potentiation was 2.5-4-fold larger than in the wild-type. Results with the H140T and H140R mutants further support the importance of a histidine residue at this position. We conclude that His-140 is critical for the action of copper, indicating that this histidine residue, but not His-241 or His-286, forms part of the inhibitory allosteric metal-binding site of the P2X4 receptor, which is distinct from the putative zinc facilitator binding site.
Journal of Biological Chemistry 10/2003; 278(38):36777-85. · 4.77 Impact Factor
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ABSTRACT: To further elucidate the modulator role of trace metals such as zinc and copper on the activity of nucleotide purinoceptors, the action of these metals was assessed during prolonged ATP applications to rat P2X(4) purinoceptors expressed in Xenopus laevis oocytes. Application of ATP for 3 min resulted in a biphasic effect; a fast transient peak was followed by a slower stable current component with similar pharmacological and biophysical characteristics. The application of 1-300 microM Cu(2+) inhibited both current components to a comparable extent; likewise, Zn(2+) facilitated to a similar degree the transient and the slower stable current components. Carnosine (Car), cysteine (Cys), histidine (His), and the metal chelator, penicillamine, prevented the inhibitory action of Cu(2+); the Zn(2+) facilitation was not prevented by neither Car nor His but by either bathophenantroline or Cys, revealing metal selectivity. While the noncompetitive Cu(2+) inhibition appears to decrease channel conductance, Zn(2+) likely increases ATP affinity independently of the activation state of the purinoceptor. These results strongly support the notion that trace metals modulate the activity of the P2X(4) purinoceptor and could become relevant during continual activity of a P2X(4) purinoceptor-containing synapse.
European Journal of Pharmacology 08/2003; 472(1-2):49-56. · 2.52 Impact Factor
