ryanodine and 3-4-dihydroxyphenylglycol

Administration of the Group 1 metabotropic glutamate receptor (mGluR) agonist (R,S)-3,5-dihydroxyphenylglycine (DHPG) facilitates ("primes") subsequent long-term potentiation (LTP) through a phospholipase C signaling cascade that may involve release of Ca2+ from the endoplasmic reticulum (ER). We investigated the intracellular calcium pathways involved in this priming effect, recording field potentials from area CA1 of rat hippocampal slices before and after high-frequency stimulation. The priming of LTP by DHPG was prevented by co-administration of cyclopiazonic acid, which depletes ER Ca2+ stores. The priming effect was also blocked by the ryanodine receptor (RYR) antagonist ryanodine (RYA, 100 microM). In contrast, a low dose of RYA (10 microM) which opens the RYR channel, by itself primed LTP. In addition to RYR activation, entry of extracellular calcium through store-operated channels appears necessary for priming, since diverse treatments known to impede store-operated channel activity completely blocked both RYA and DHPG priming effects. Thus, RYR activation plays a critical role in the priming of LTP by Group 1 mGluRs, and this effect is coupled to the entry of extracellular calcium, probably through store-operated calcium channels.

Topics: Animals; Boron Compounds; Calcium; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Drug Interactions; Electric Stimulation; Enzyme Inhibitors; Excitatory Amino Acid Agonists; Excitatory Postsynaptic Potentials; Hippocampus; In Vitro Techniques; Indoles; Long-Term Potentiation; Male; Methoxyhydroxyphenylglycol; Nitriles; Patch-Clamp Techniques; Rats; Rats, Sprague-Dawley; Ryanodine; Ryanodine Receptor Calcium Release Channel; Tyrphostins

Group I metabotropic glutamate receptors (mGluRs) are positively coupled to phospholipase C (PLC) via Galphaq-proteins and are expressed in the medium-sized projection neurons of striatum. To characterize the group I mGluR/PLC-sensitive modulation of intracellular Ca2+ ([Ca2+]i) signalling, primary neuronal cultures were prepared from the striatum of E19 rat embryos or neonatal day-1 rat pups. Cytoplasmic Ca2+ signals were examined with fura-2/AM at a signal cell level. After 17-18 days in culture, a profound Ca2+ response consisting of two phases was induced in cultured striatal neurons following bath application of the selective group I agonist, 3,5-dihydroxyphenylglycine (DHPG). The [Ca2+]i elevation was concentration- and time-dependent, and was blocked by coexposure to the group I antagonist, N-phenyl-7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxamide (PHCCC), or the PLC inhibitor, U-73122, but not to the group II/III antagonist (RS)-alpha-methylserine-O-phosphate monophenyl ester (MSOPPE). A series of further pharmacological studies demonstrated that the initial spike-like transient was dependent on intracellular Ca2+ mobilization through 1,4,5-triphosphate-sensitive stores, and the second long-lasting rise was dependent on extracellular Ca2+ influx through N-methyl-d-aspartate (NMDA) receptors and especially L-type voltage-operated Ca2+ channels. Lastly, using an immediate early gene c-fos as a report of inducible gene expression, the resultant [Ca2+]i elevation contributes to DHPG-stimulated c-fos mRNA and Fos protein expression in striatal neurons as revealed by quantitative in situ hybridization and immunocytochemistry, respectively. These results demonstrate that group I mGluRs are able to affect Ca2+ homeostasis at multiple levels and trigger Ca2+-sensitive gene transcription in striatal neurons.

Topics: Animals; Animals, Newborn; Benzopyrans; Calcium Channel Blockers; Calcium Signaling; Cell Count; Cells, Cultured; Corpus Striatum; Dose-Response Relationship, Drug; Drug Interactions; Embryo, Mammalian; Enzyme Inhibitors; Fura-2; Gene Expression; Genes, fos; Immediate-Early Proteins; Immunohistochemistry; In Situ Hybridization; Methoxyhydroxyphenylglycol; Neurons; Nifedipine; Proto-Oncogene Proteins c-fos; Rats; Receptors, Metabotropic Glutamate; Ryanodine; Time Factors

The modulatory action of calcium (Ca2+) released from intracellular stores on GABAA receptor-mediated current was investigated in wide-field amacrine cells isolated from the teleost, Morone chrysops, retina. Caffeine, ryanodine or inositol 1,4,5-triphosphate (IP3) markedly inhibited the GABAA current by elevating [Ca2+]i. The inhibition resulted from the activation of a Ca2+--> Ca2+/calmodulin --> calcineurin cascade. Long (>12 s) exposure to glutamate mimicked the caffeine effect, i.e. it inhibited the GABAA current by elevating [Ca2+]i through mGluR1 receptor activation and consequent IP3 generation. This pathway provides a 'timed' disinhibitory mechanism to potentiate excitatory postsynaptic potentials in wide-field amacrine cells. It occurs as a result of the suppression of GABA-mediated conductances as a function of the duration of presynaptic excitatory input activity. This is much like some forms of long-term potentiation in the central nervous system. In a local retinal circuit this will selectively accentuate particular excitatory inputs to the wide-field amacrine cell. Similar to other neural systems, we suggest that activity-dependent postsynaptic disinhibition is an important feature of the signal processing in the inner retina.

Topics: alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; Amacrine Cells; Animals; Anticoagulants; Bicuculline; Caffeine; Calcium; Calcium Channels; Carps; Cells, Cultured; Central Nervous System Stimulants; Chelating Agents; Dose-Response Relationship, Drug; Drug Interactions; Egtazic Acid; Electric Conductivity; Enzyme Inhibitors; Excitatory Amino Acid Agonists; Extracellular Space; GABA Antagonists; gamma-Aminobutyric Acid; Glutamic Acid; Heparin; Immunohistochemistry; Inositol 1,4,5-Trisphosphate; Inositol 1,4,5-Trisphosphate Receptors; Kainic Acid; Membrane Potentials; Methoxyhydroxyphenylglycol; Organophosphorus Compounds; Patch-Clamp Techniques; Receptors, Cytoplasmic and Nuclear; Receptors, GABA-A; Receptors, Metabotropic Glutamate; Retina; Ryanodine

Astrocytes in the rat thalamus display spontaneous [Ca(2+)](i) oscillations that are due to intracellular release, but are not dependent on neuronal activity. In this study we have investigated the mechanisms involved in these spontaneous [Ca(2+)](i) oscillations using slices loaded with Fluo-4 AM (5 microM) and confocal microscopy. Bafilomycin A1 incubation had no effect on the number of spontaneous [Ca(2+)](i) oscillations indicating that they were not dependent on vesicular neurotransmitter release. Oscillations were also unaffected by ryanodine. Phospholipase C (PLC) inhibition decreased the number of astrocytes responding to metabotropic glutamate receptor (mGluR) activation but did not reduce the number of spontaneously active astrocytes, indicating that [Ca(2+)](i) increases are not due to membrane-coupled PLC activation. Spontaneous [Ca(2+)](i) increases were abolished by an IP3 receptor antagonist, whilst the protein kinase C (PKC) inhibitor chelerythrine chloride prolonged their duration, indicating a role for PKC and inositol 1,4,5,-triphosphate receptor activation. BayK8644 increased the number of astrocytes exhibiting [Ca(2+)](i) oscillations, and prolonged the responses to mGluR activation, indicating a possible effect on store-operated Ca(2+) entry. Increasing [Ca(2+)](o) increased the number of spontaneously active astrocytes and the number of transients exhibited by each astrocyte. Inhibition of the endoplasmic reticulum Ca(2+) ATPase by cyclopiazonic acid also induced [Ca(2+)](i) transients in astrocytes indicating a role for cytoplasmic Ca(2+) in the induction of spontaneous oscillations. Incubation with 20 microM Fluo-4 reduced the number of astrocytes exhibiting spontaneous increases. This study indicates that Ca(2+) has a role in triggering Ca(2+) release from an inositol 1,4,5,-triphosphate sensitive store in astrocytes during the generation of spontaneous [Ca(2+)](i) oscillations.

Topics: 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; Aniline Compounds; Animals; Animals, Newborn; Anti-Infective Agents, Local; Astrocytes; Boron Compounds; Caffeine; Calcium; Calcium Channel Agonists; Calcium Signaling; Dioxolanes; Drug Interactions; Enzyme Inhibitors; In Vitro Techniques; Methoxyhydroxyphenylglycol; Microscopy, Confocal; Patch-Clamp Techniques; Purines; Rats; Ryanodine; Thalamus; Thimerosal; Xanthenes

The response of the semicircular canal (SCC) to the group I mGluR-selective agonist dihydroxyphenylglycine (DHPG; 300 microM) - facilitation of afferent discharge rate - was dose-dependently reduced by the phospholipase C inhibitor U-73122 (1-100 microM; IC(50): 22 microM), the smooth endoplasmic reticulum Ca(++) ATPase inhibitor thapsigargin (100 nM-3 microM; IC(50): 500 nM), and xestospongin C (100 pM-1 microM; IC(50): 11 nM), an inositol trisphosphate receptor (IP(3)R) antagonist. Ryanodine, a modulator of Ca(++)-induced Ca(++) release, biphasically facilitated, then suppressed this response (1 nM-1 mM; approximate IC(50): 50 microM). 5 mM caffeine increased the amplitude (34.6+/-13.4%) and duration (453+/-169.8%; n=4) of the response of the SCC to DHPG, while 50 mM caffeine eliminated this response (n=2). The protein kinase C inhibitor bisindolylmaleimide I-HCl (10-100 microM; n=3) and the cyclic-ADP ribose antagonist 8-Br-cyclic-ADP ribose (1-10 microM; n=3) had no effect on the response of the SCC to DHPG. These data suggest that the increase in transmitter release following activation of group I mGluRs on vestibular hair cells is associated with intracellular Ca(++) release from both IP(3)-sensitive and ryanodine/caffeine-sensitive intracellular Ca(++) stores. Such positive feedback on transmitter release may serve to enhance the contrast between the spontaneous and stimulus-evoked modes of hair cell transmitter release, thereby optimizing signal discrimination at the synapse between hair cells and vestibular afferent fibers.

Topics: Animals; Auditory Pathways; Calcium Signaling; Electrophysiology; Hair Cells, Auditory; In Vitro Techniques; Macrocyclic Compounds; Methoxyhydroxyphenylglycol; Neurotransmitter Agents; Oxazoles; Rana pipiens; Receptors, Metabotropic Glutamate; Ryanodine; Semicircular Canals; Thapsigargin

Experimental and computational techniques have been used to investigate the group I metabotropic glutamate receptor (mGluR)-mediated increase in the frequency of spinal cord network activity underlying locomotion in the lamprey. Group I mGluR activation potentiated the amplitude of NMDA-induced currents in identified motoneurons and crossed caudally projecting network interneurons. Group I mGluRs also potentiated NMDA-induced calcium responses. This effect was blocked by a group I mGluR-specific antagonist, but not by blockers of protein kinase A, C, or G. The effect of group I mGluRs activation was also tested on NMDA-induced oscillations known to occur during fictive locomotion. Activation of these receptors increased the duration of the plateau phase and decreased the duration of the hyperpolarizing phase. These effects were blocked by a group I mGluR antagonist. To determine its role in the modulation of NMDA-induced oscillations and the locomotor burst frequency, the potentiation of NMDA receptors by mGluRs was simulated using computational techniques. Simulating the interaction between these receptors reproduced the modulation of the plateau and hyperpolarized phases of NMDA-induced oscillations, and the increase in the frequency of the locomotor rhythm. Our results thus show a postsynaptic interaction between group I mGluRs and NMDA receptors in lamprey spinal cord neurons, which can account for the regulation of the locomotor network output by mGluRs.

Topics: Animals; Biological Clocks; Calcium; Cells, Cultured; Computer Simulation; GTP-Binding Proteins; Lampreys; Locomotion; Methoxyhydroxyphenylglycol; N-Methylaspartate; Nerve Net; Neural Networks, Computer; Neurons; Patch-Clamp Techniques; Protein Kinase Inhibitors; Receptors, AMPA; Receptors, Kainic Acid; Receptors, Metabotropic Glutamate; Receptors, N-Methyl-D-Aspartate; Ryanodine; Spinal Cord; Synaptic Transmission