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J Sánchez-Prieto

Publications and source records attributed to J Sánchez-Prieto.

At least 19 recordsLinked to original sources
PubMed2005

Inhibition of N- and P/Q-type Ca2+ channels by cannabinoid receptors in single cerebrocortical nerve terminals.

Since cannabinoid receptors inhibit excitatory synaptic transmission by reducing glutamate release, we have examined whether this might occur through the direct inhibition of presynaptic Ca2+ channels. In cerebrocortical nerve terminals, activation of cannabinoid receptors with WIN55,212-2 reduces the KCl-evoked release of glutamate. However, this inhibition is attenuated when N- and P/Q-type Ca2+ channels are blocked. Through Ca2+ imaging in single nerve terminals, we found that WIN55,212-2 reduced the influx of Ca2+ both in nerve terminals that contain N-type Ca2+ channels and those that contain P/Q-type Ca2+ channels. Thus, cannabinoid receptors modulate the two major Ca2+ channels coupled to glutamate release in the cerebral cortex.

Animals↗
PubMed2004

Dual signaling by mGluR5a results in bi-directional modulation of N-type Ca2+ channels.

We have studied how N-type Ca2+ channels are modulated by the metabotropic glutamate receptor 5a (mGluR5a) in Xenopus oocytes. Stimulation of the receptor with glutamate initiated two parallel responses, a rapid inhibition followed by an upregulation of the Ca2+ current. Although a subsequent stimulation did not upregulate the Ca2+ current, it did still produce a reduction in the amplitude of the current. The upregulation of Ca2+ channels was prevented by the protein kinases inhibitor staurosporine and it was mimicked by the activation of PKC with phorbol esters. In contrast, the inhibition of the Ca2+ current was insensitive to staurosporine. These results show that mGluR5a exerts a bi-directional influence on Ca2+ channels, which may explain how group I mGluRs facilitate and inhibit glutamate release at central synapses.

Animals↗
PubMed2001

An activity-dependent switch from facilitation to inhibition in the control of excitotoxicity by group I metabotropic glutamate receptors.

Activation of group I metabotropic glutamate receptors (mGlu1 or -5 receptors) is known to either enhance or attenuate excitotoxic neuronal death depending on the experimental conditions. We have examined the possibility that these receptors may switch between two different functional modes in regulating excitotoxicity. In mixed cultures of cortical cells, the selective mGlu1/5 agonist, 3,5-dihydroxyphenylglycine (DHPG), amplified neurodegeneration induced by a toxic pulse of NMDA. This effect was observed when DHPG was either combined with NMDA or transiently applied to the cultures prior to the NMDA pulse. However, two consecutive applications of DHPG consistently produced neuroprotection. Similar effects were observed with DHPG or quisqualate (a potent agonist of mGlu1/5 receptors) in pure cultures of cortical neurons virtually devoid of astrocytes. In cultures of hippocampal pyramidal neurons, however, only protective effects of DHPG were seen suggesting that, in these particular cultures, group I mGlu receptors were endogenously switched into a "neuroprotective mode". The characteristics of the activity-dependent switch from facilitation to inhibition were examined in mixed cultures of cortical cells. The switch in the response to DHPG was observed when the two applications of the drug were separated by an interval ranging from 1-45 min, but was lost when the interval was extended to 90 min. In addition, this phenomenon required the initial activation of mGlu5 receptors (as indicated by the use of subtype-selective antagonists) and was mediated by the activation of protein kinase C. We conclude that group I mGlu receptors are subjected to an activity-dependent switch in regulating excitotoxic neuronal death and, therefore, the recent "history" of these receptors is critical for the response to agonists or antagonists.

Animals↗
PubMed2000

Protein phosphatase 2B inhibitors mimic the action of arachidonic acid and prolong the facilitation of glutamate release by group I mGlu receptors.

We have addressed the role of arachidonic acid in the facilitation of glutamate release by group I metabotropic glutamate (mGlu) receptors. The activation of these receptors with the specific agonist 3,5-dihydroxyphenylglycine (DHPG) failed to enhance the cumulative Ca(2+)-dependent release of glutamate evoked by a 5 min depolarization with 4-aminopyridine, in the absence but not in the presence of arachidonic acid. However, DHPG, in the absence of arachidonic acid, transiently enhanced diacylglycerol levels, transiently potentiated 4AP-evoked depolarization, and significantly enhanced the fast but not the slow component of glutamate release observed after prolonged stimulations of nerve terminals. Further evidence that DHPG was able to initiate release facilitation in the absence of arachidonic acid was obtained in experiments where the protein phosphatase 2B (cyclosporine A and cypermethrine) but not protein phosphatase 1 or 2A inhibitors (okadaic acid and calyculin A) facilited glutamate release to a maximal extent comparable to that induced by arachidonic acid. We conclude that an active protein phosphatase 2B (calcineurin) dephosphorylates the presynaptic target/s responsible for facilitation of glutamate release.

4-Aminopyridine↗
PubMed2000

Protein phosphatase 1 and 2A inhibitors prolong the switch in the control of glutamate release by group I metabotropic glutamate receptors: characterization of the inhibitory pathway.

We have addressed the role of protein phosphatases (PPs) in the modulation of the switch in glutamate release observed after repetitive stimulation of group I metabotropic glutamate receptors (mGluRs). In cerebrocortical nerve terminals the agonist (S:)-3, 5-dihydroxyphenylglycine facilitated evoked glutamate release. However, a second stimulation, 5 min later, reduced rather than facilitated this release. This switch in the control of glutamate release was reversed when a 30-min interval was left between stimulations. Inhibition of the endogenous PPs, PP1 and PP2A, with calyculin A and okadaic acid prevented the recovery of the facilitatory response and maintained the receptor permanently coupled to the inhibitory pathway. The inhibitors of PP2B, cyclosporin A and cypermethrine, had no effect. The inhibition of glutamate release was insensitive to pertussis toxin and was the result of the loss of the release component coupled to N-type Ca(2+) channels. This inhibitory action was suppressed by addition of the protein kinase C activator 4beta-phorbol 12,13-dibutyrate. We conclude that the balance between protein kinase and phosphatase activity at the nerve terminal plays a key role in accommodating the modulation of glutamate release by group I mGluRs.

4-Aminopyridine↗
PubMed1999

Presynaptic group 1 metabotropic glutamate receptors may contribute to the expression of long-term potentiation in the hippocampal CA1 region.

In this study, we investigated the possible contribution of presynaptic group 1 metabotropic glutamate receptor activation to changes in synaptic efficacy by means of analysis of glutamate release in hippocampal synaptosomes. Data were interpreted in the context of group 1 metabotropic glutamate receptor involvement in synaptic plasticity in the CA1 region of freely moving rats. In synaptosomes, 3,5-dihydroxyphenylglycine enhanced diacylglycerol formation and facilitated vesicular Ca(2+)-dependent glutamate release, whereas trans-azetidine-2,4-dicarboxylic acid had no effect on these processes. Trans-azetidine-2,4-dicarboxylic acid enhanced glutamate release, but in a Ca(2+)-independent manner. This effect was mimicked by the L-glutamate uptake inhibitor L-trans-pyrrolidine-2,4-dicarboxylic acid. (R,S)-alpha-Methyl-4-carboxyphenylglycine blocked the effects of 3,5-dihydroxyphenylglycine, but not trans-azetidine-2,4-dicarboxylic acid in synaptosomes. Short-term potentiation (100 Hz, three bursts of 10 stimuli, 0.1 ms stimulus duration, 10 s interburst interval) was induced in the CA1 region in vivo. The metabotropic glutamate receptor agonist 1S,3R-aminocyclopentane-2,3-dicarboxylic acid, or the group 1 metabotropic glutamate receptor agonists, 3,5-dihydroxyphenylglycine and trans-azetidine-2,4-dicarboxylic acid, dose-dependently facilitated short-term potentiation into long-term potentiation, which lasted > 24 h. The facilitation was inhibited by the metabotropic glutamate receptor antagonist, (R,S)-alpha-methyl-4-carboxyphenylglycine, and the group 1 metabotropic glutamate receptor antagonist, (S)-4-carboxy-phenylglycine, but not by the group 2 metabotropic glutamate receptor antagonist, (R,S)-alpha-methylserine-O-phosphate monophenyl ester. L-Trans-pyrrolidine-2,4-dicarboxylic acid dose-dependently facilitated short-term potentiation into long-term potentiation, which lasted < 4 h. These data suggest that activation of group 1 metabotropic glutamate receptors results in presynaptic modulation of glutamate release. This effect may contribute to group 1 metabotropic glutamate modulation of the expression of long-term potentiation in vivo.

Animals↗
PubMed1998

Functional switch from facilitation to inhibition in the control of glutamate release by metabotropic glutamate receptors.

We have investigated the role of metabotropic glutamate receptors linked to phosphoinositide hydrolysis in the control of glutamate release in cerebrocortical nerve terminals. The activation of these receptors with the agonist 3,5-dihydroxyphenylglycine enhanced intra-synaptosomal diacylglycerol and facilitated both the depolarization-induced increase in the cytosolic free Ca2+ concentration and the release of glutamate. However, 5 min after receptor activation, a second stimulation of the pathway with the agonist failed to produce diacylglycerol and to facilitate glutamate release. Interestingly, during the period in which the diacylglycerol response was desensitized, a strong agonist-induced inhibition of Ca2+ entry and glutamate release was observed. This change in the presynaptic effects of 3,5-dihydroxyphenylglycine is reversible since 30 min after the first stimulation, the agonist-induced inhibition of release disappeared, whereas both the production of diacylglycerol and the facilitation of glutamate release were recovered. The tonic elevation of the extracellular glutamate concentration from basal levels (0.8 microM) up to 5 microM also produced the switch from facilitation to inhibition in the receptor response. The existence of this activity-dependent switch in the presynaptic control of glutamate release suggests that release facilitation is limited to conditions under which an appropriate clearance of synaptic glutamate exists, probably to prevent the neurotoxic accumulation of glutamate in the synapse.

Animals↗
PubMed1998

The metabotropic glutamate receptor 1 is not involved in the facilitation of glutamate release in cerebrocortical nerve terminals.

In this study we have addressed the identification of the metabotropic glutamate receptor (mGluR) involved in the facilitation of glutamate release in nerve terminals from the cerebral cortex. mGluR1 and 5 are coupled to phosphoinositide hydrolysis and the activation of these receptors with the specific agonist 3,5-dihydroxyphenylglycine (DHPG) enhances the release of glutamate. We have examined whether mGluR1 is responsible for this modulatory effect by preparing nerve terminals from mGluR 1 deficient mice. The Ca2+-dependent glutamate release evoked by a submaximal depolarization is enhanced by the agonist DHPG in nerve terminals from both wild and mutant mice. This result is consistent with the finding that the mGluR agonist also induces a similar increase in the levels of diacylglycerol (DAG) in the nerve terminals from wild and mutant mice. Moreover, the activity-dependent switch from facilitation to inhibition of release, observed when a second stimulation of the receptor is applied shortly after (5 min) the first pulse, was also observed in the mutant mice. These results indicate therefore, that the facilitation of glutamate release is unlikely to be due to the activation of mGluR1 but related to another phosphoinositide coupled mGluR.

Adenosine↗
PubMed1998

Switch from facilitation to inhibition of excitatory synaptic transmission by group I mGluR desensitization.

We have explored whether the desensitization of metabotropic glutamate receptors (mGluRs) coupled to phosphoinositide hydrolysis affects the role that they play in modulating glutamate release. In hippocampal nerve terminals, the agonist 3,5-dihydroxyphenylglycine (DHPG) facilitated evoked glutamate release, but a second stimulation 5 min later reduced rather than facilitated release. After a 30 min interval between stimulations, DHPG again facilitated glutamate release. In hippocampal slices, DHPG caused an inhibition of excitatory postsynaptic currents (EPSCs) recorded from CA1 neurons. However, when the effects of ambient glutamate were prevented, mGluR activation initially induced a facilitation of synaptic transmission, followed by an inhibition. We conclude that group I mGluRs have a dual action on glutamate release, switching from facilitatory to inhibitory upon receptor desensitization triggered by low concentrations of glutamate.

Animals↗
PubMed1997

Modulation of glutamate release by a nitric oxide/cyclic GMP-dependent pathway.

The mechanism by which changes in cyclic GMP (cGMP) regulate glutamate release was investigated in rat cerebrocortical nerve terminals. The elevation of cGMP levels by inhibition of cGMP-phosphodiesterase with 2-o-propoxy-phenyl-8-azapurin-6-one (zaprinast) reduced the Ca(2+)-dependent glutamate release evoked by depolarization with 30 mM KCl or 1 mM 4-aminopyridine. The nitric oxide (NO) donor S-nitroso-N-acetylpenicillamine also enhanced cGMP and reduced glutamate release. In addition, the membrane-permeable analogs 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br-cGMP) and N,2'-o-dibutyrylguanosine (dbcGMP) at 10 microM also mimic glutamate release inhibition. The reduction in glutamate release was observed with no modifications in the ATP/ADP ratio, and was reversed in the presence of the protein kinases inhibitor [N-[2-(methylamino)ethyl]-5-isoquinoline sulfonamide, HCl] (H-8). Interestingly, higher concentrations of dbcGMP (1 mM) abolished the inhibition observed with low concentrations although no facilitation was observed. This finding seems to indicate the existence of a dual role for cGMP in the control of glutamate exocytosis.

Adenosine Triphosphate↗
PubMed1997

Presynaptic modulation of glutamate release targets different calcium channels in rat cerebrocortical nerve terminals.

We have studied which type/s of Ca2+-channel/s support glutamate exocytosis and its modulation by presynaptic receptors in cerebrocortical nerve terminals. Depolarization of nerve terminals with 30 mM KCl induced a Ca2+-dependent release of 3.64 +/- 0.25 nmol/mg of protein. The addition of either 2 microM omega-conotoxin-GVIA or 200 nM omega-agatoxin-IVA reduced the KCl-evoked release by 47.7 +/- 3.5% and 70.4 +/- 8.9% respectively, and by 85.7 +/- 4.1% when both toxins were co-applied. The activation of adenosine A1 receptors with N6-cyclohexyladenosine or the activation of metabotropic glutamate receptors with L(+)-2-amino-4-phosphonobutyrate inhibited the KCl-evoked release by 41.0 +/- 5.9 and 54.3 +/- 10% respectively. The extent of these inhibitions was not altered by the prior addition of 2 microM omega-conotoxin-GVIA but they were significantly enhanced when omega-agatoxin-IVA was added together with the adenosine A1 receptor agonist or the metabotropic glutamate receptor agonist, suggesting that omega-conotoxin-GVIA-sensitive and not omega-agatoxin-IVA-sensitive Ca2+-channels are involved in the action of these inhibitory receptors. By contrast, the facilitation of glutamate release that follows the activation of the protein kinase C, either with phorbol esters or with the stimulation of phospholipase C-linked metabotropic receptors, was expressed by both omega-conotoxin-GVIA-sensitive and omega-agatoxin-sensitive Ca2+-channels. It is concluded that different Ca2+-channels support the modulation of glutamate release by presynaptic receptors.

Adenosine↗
PubMed1996

cAMP-dependent facilitation of glutamate release by beta-adrenergic receptors in cerebrocortical nerve terminals.

We have investigated the presence of a cAMP-protein kinase A-dependent pathway in cerebrocortical nerve terminals and its role in the modulation of glutamate release. The activation of adenylyl cyclase with forskolin enhances intrasynaptosomal cAMP and induces Ca2+-dependent glutamate release. The membrane permeant analogue dibutyryl cAMP mimics this facilitatory effect, whereas the inactive compound 1,9-dideoxyforskolin is without effect. This cAMP-induced facilitation is consistent with the induction of spontaneous action potentials that are abolished by the Na+ channel blocker tetrodotoxin and by reducing nerve terminal excitability with arachidonic acid. We have also demonstrated that a beta-adrenergic receptor is linked to this pathway because isoproterenol increases cAMP levels and glutamate release, and both actions are antagonized by the receptor antagonist propanolol and the protein kinase A inhibitors H89 and 8-chloroadenosine 3',5'-monophosphorothioate ((Rp)-isomer). The finding that the increase in cytoplasmic free Ca2+ concentration induced by synaptic activity reduces the concentration of agonist required to maximally activate adenylyl cyclase suggests that this enzyme may act as a coincidence detector, integrating glutamatergic neurotransmission and noradrenaline release.

Animals↗
PubMed1996

Glutamate release evoked by glutamate receptor agonists in cultured chick retina cells: modulation by arachidonic acid.

We studied the effect of ionotropic glutamate receptor agonists on the release of endogenous glutamate or of [3H]D-aspartate from reaggregate cultures (retinospheroids) or from monolayer cultures of chick retinal cells, respectively. Kainate increased the fluorescence ratio of the Na+ indicator SBFI and stimulated a dose-dependent release of glutamate in low (0.1 mM) Ca2+ medium, as measured using a fluorometric assay. Under the same experimental conditions, the release evoked by N-methyl-D-aspartate (NMDA; 400 microM) was about half of that evoked by the same kainate concentration; alpha-amino-3-hydroxy-5-methyl-4-isoxasolepropionic acid (AMPA; 400 microM) did not trigger a significant response. In the presence of 1 mM CaCl2, all of the agonists increased the [Ca2+]i, as determined with the fluorescence dye Indo-1, but the glutamate release evoked by NMDA and kainate was significantly lower than that measured in 0.1 mM CaCl2 medium. Inhibition by Ca2+ of the kainate-stimulated release of glutamate was partially reversed by the phospholipase A2 inhibitor oleiloxyethyl phosphorylcholine (OPC), suggesting that the effect was mediated by the release of arachidonic acid, which inhibits the glutamate carrier. Accordingly, kainate, NMDA, and AMPA stimulated a Ca(2+)-dependent release of [3H]arachidonic acid, and the direct addition of the exogenous fatty acid to the medium decreased the release of glutamate evoked by kainate in low (0.1 mM) CaCl2 medium. In monolayer cultures, we showed that NMDA, kainate, and AMPA also stimulated the release of [3H]D-aspartate, but in this case release in the presence of 1 mM CaCl2 was significantly higher than that evoked in media with no added Ca2+. The ranking order of efficacy for stimulation of Ca(2+)-dependent release of [3H]D-aspartate was NMDA > > kainate > AMPA.

Animals↗
PubMed1996

On-line detection of glutamate release from cultured chick retinospheroids.

A continuous fluorometric assay was adapted to measure the release of endogenous glutamate from cultured chick retinospheroids. The results obtained with this technique are compared with the release of [3H]D-aspartate from monolayer cultures of chick retina cells. It is shown that although excitatory amino acids may be released in a Ca(2+)-dependent manner, most of the neurotransmitter release from cultured retina cells occurs by reversal of the glutamate transporter. The presence of extracellular Ca2+ may actually inhibit glutamate release by the cells present in the retinospheroids, or the [3H]D-aspartate release by cells in monolayers, when veratridine is the depolarizing agent.

ATP-Binding Cassette Transporters↗
PubMed1996

Presynaptic receptors and the control of glutamate exocytosis.

When a typical glutamate-containing neurone fires, an action potential is propagated down the branching axon through more than a thousand varicosities. At each of these release sites the probability that a synaptic vesicle will be exocytosed into the synaptic cleft is individually controlled by means of presynaptic receptors: autoreceptors responding by positive or negative feedback to previously released transmitter, or heteroreceptors under the influence of other neurotransmitters or modulators. The simplest system in which to investigate presynaptic modulation is the isolated nerve terminal or synaptosome; studies with this preparation have revealed a complex interplay of signal-transduction pathways.

Animals↗
PubMed1996

Two components of glutamate exocytosis differentially affected by presynaptic modulation.

The total Ca(2+)-dependent release of glutamate induced by depolarization of cerebrocortical nerve terminals with KCl was analyzed into a fast and a slow component. The fast component exhibited a decay time of < 1 s and accounted for 0.95 +/- 0.10 nmol of glutamate, whereas the slow component, which exhibited a decay time of 52 +/- 7 s, accounted for the release of 2.48 +/- 0.19 nmol of glutamate. These two components were differentially affected by the Ca2+ chelator BAPTA, the divalent cation Sr2+, or the botulinum neurotoxin A. The adenosine A1 receptor agonist N6-cyclohexyladenosine strongly reduced the fast component without altering the slow component. In contrast, the inhibitory effect of arachidonic acid and the facilitatory action of the metabotropic glutamate receptor agonist (1S, 3R)-1-aminocyclopentane-1, 3-dicarboxylic acid were observed as a decrease and an increase, respectively, in the two components. It is concluded, first, that the fast and slow components correspond to the release of docked and mobilized vesicles, respectively, and second, that presynaptic modulation more significantly alters the fast component of release.

Animals↗
PubMed1996

[3H]heparin binding in boar spermatozoa: characterization and correlation with routine semen quality parameters.

A simple technique has been established for the purpose of characterizing heparin binding to boar sperm. Binding experiments were performed using [3H]heparin and extended boar semen. [3H]Heparin binding to boar sperm was effectively displaced by increasing concentrations of heparin. [3H]Heparin binding was linear at least between 50,000 and 10(6) spermatozoa and was stable for at least 120 min. Binding was sensitive to fucoidan, chondroitin sulfate B, chondroitin sulfate C, and chondroitin sulfate A, while keratan sulfate had only a marginal effect on binding. The [3H]heparin binding was saturable. Assuming a 13,000 M(r) for the [3H]heparin used, binding to boar spermatozoa showed an apparent equilibrium dissociation constant (Kd) of 23.6 +/- 2.5 nM and a maximum binding capacity (Bmax) of 6.0 +/- 1.1 pmol/10(6) spermatozoa (average values from 6 boars, means +/- SEM). Interejaculate variations in binding parameters were dependent on the male. Thus, with respect to Bmax variation, 4 of 6 boars studied exhibited an interejaculate coefficient of variation of less than 0.33 (0.09, 0.11, 0.11, and 0.29, respectively, for 3 consecutive ejaculates), while in the case of Kd interejaculate variation, only 2 of the 6 boars studied showed acceptable variation coefficients (0.16 and 0.28). No seasonal effect was observed in either of the binding parameters, with the variations following boar-specific patterns. Kd and Bmax intermean differences for different boars during the course of the study period (April-June) were not significant (p > 0.05). Correlations of mean boar binding parameters (Kd and Bmax) with conventional semen quality parameters showed a correlation between Bmax values and those of the osmotic resistance test (r = 0.8990, p < 0.01), normal acrosomes (r = 0.7946, p < 0.05), and bended tails (r = -0.8632, p < 0.02). Kd values were correlated with cytoplasmic distal droplet (r = -0.7992, p < 0.05). The glycosaminoglycans and polysulfated boar sperm binding sites reported in the present work should be regarded as binding sites until further studies elucidate their physiological role. The results obtained suggest that this technique be given a role as a research tool.

Animals↗