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T G Smart

Publications and source records attributed to T G Smart.

At least 55 records · Page 3Linked to original sources
PubMed1994

Modulation of long-term potentiation in rat hippocampal pyramidal neurons by zinc.

The phenomenon of long-term potentiation is frequently promulgated as an example of learning and memory mechanisms at the synaptic level in the mammalian central nervous system. In the CA3 region of the hippocampus there is an abundance of zinc, which is located in presynaptic mossy fibre nerve terminals. Stimulation of these fibres can cause the release of zinc, which interacts with excitatory amino acid receptors and may therefore modulate long-term potentiation. We now demonstrate in CA1 and CA3 neurons that zinc (100-300 microM) enhances non-N-methyl-D-aspartate-receptor-mediated responses whilst reducing excitatory synaptic transmission and inhibiting long-term potentiation. However, by using zinc-chelating agents, endogenously released zinc following high-frequency stimulation in the stratum lucidum does not appear to have any modulatory role in excitatory synaptic transmission and long-term potentiation. These results indicate that an increase in the level of extracellular zinc can limit excitatory synaptic transmission in the CA1 or CA3 region and further suggests that pathologies that can be related to excessive levels of endogenous zinc may have implications for synaptic plasticity in CA3 neurons.

Animals↗
PubMed1994

Regulation of GABAA receptor function by protein kinase C phosphorylation.

GABAA receptors possess consensus sequences for phosphorylation by PKC that are located on the presumed intracellular domains of beta and gamma 2 subunits. PKC phosphorylation sites were analyzed using purified receptor subunits and were located on up to 3 serine residues in beta 1 and gamma 2 subunits. The role of phosphorylation in receptor function was studied using recombinant receptors expressed in kidney cells and Xenopus oocytes and was compared with native neuronal GABAA receptors. For recombinant and native GABAA receptors, PKC phosphorylation caused a reduction in the amplitudes of GABA-activated currents without affecting the time constants for current decay. Selective site-directed mutagenesis of the serine residues reduced the effects of phorbol esters and revealed that serine 343 in the gamma 2 subunit exerted the largest effect on the GABA-activated response. These results indicate that PKC phosphorylation can differentially modulate GABAA receptor function.

Alanine↗
PubMed1994

m-sulphonate benzene diazonium chloride: a novel GABAA receptor antagonist.

A previously identified irreversible affinity label for the gamma-aminobutyric acid (GABA) binding site in rat brain membranes, m-sulphonate benzene diazonium chloride (MSBD), was characterized in functional studies using patch clamp and two-electrode voltage clamp recording techniques. MSBD did not exhibit any agonist activity on native GABAA receptors in cultured sympathetic ganglionic neurones but acted as an antagonist of GABA-induced membrane currents. Recombinant GABAA receptors composed of alpha 1, beta 1 and gamma 2S subunits were expressed in Xenopus oocytes following microinjection with cDNAs. Equilibrium dose-response curve analyses established that MSBD was a partially reversible, apparently non-competitive GABAA receptor antagonist. The IC50 for MSBD was estimated from an inhibition curve as 87 +/- 3 microM. In addition, the onset and recovery from MSBD-induced inhibition was independent of GABAA receptor activation. The relatively simple structure of this novel GABAA receptor antagonist, MSBD, is compared with known agonists and antagonists at the GABAA receptor. MSBD may be a useful pharmacological tool which could be used to deduce further information about the structure and function of agonist and antagonist binding sites on the GABAA receptor.

Animals↗
PubMed1994

Species-dependent functional properties of non-NMDA receptors expressed in Xenopus laevis oocytes injected with mammalian and avian brain mRNA.

1. Species-dependent variation in the functional properties of non-NMDA receptors was investigated by intracellular recording in Xenopus laevis oocytes injected with rat, chick and calf brain mRNA. 2. In all mRNA-injected oocytes, kainic acid (KA), domoic acid (Dom) and 5-bromowillardiine (BrW) evoked large, maintained membrane currents, in contrast to the smaller, desensitizing responses elicited by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), quisqualic acid (QA) and L-glutamic acid (L-Glu). Dose-response curves for KA in oocytes injected with calf (EC50 = 96.4 +/- 12.3 microM; mean +/- s.e. mean), chick (87.0 +/- 8.9 microM) or rat (88.7 +/- 4.3 microM) brain mRNA were similar. 3. Current-voltage (I-V) relationships determined with KA inwardly rectified in oocytes injected with calf or chick mRNA; whereas, outward rectification was observed in oocytes injected with rat brain mRNA. 4. In oocytes injected with rat brain mRNA, AMPA antagonized responses evoked by KA in a competitive manner. The absolute amplitudes of KA and AMPA responses in the same oocytes were significantly correlated, which is consistent with both agonists acting on the same receptor-ionophore complex. 5. In contrast, in oocytes injected with calf or chick brain mRNA, AMPA (QA and L-Glu) antagonized the response evoked by KA in a non-competitive manner. The response amplitudes of KA compared to AMPA, QA or L-Glu in the same oocytes were not correlated suggesting discrete receptor-ionophores. 6. This study favours the existence of distinct non-NMDA receptor subtypes that are equi-sensitive to KA. The expressed receptors from different species of mRNA may be distinguished by their voltage sensitivities and the type of antagonism exerted by AMPA on KA-activated responses. Our observations may reflect further heterogeneity of non-NMDA receptors in the central nervous system of different vertebrate species.

Animals↗
PubMed1993

Interaction of zinc with ionotropic and metabotropic glutamate receptors in rat hippocampal slices.

The actions of zinc on ionotropic and metabotropic glutamate receptors were studied using intracellular recording in acutely prepared adult rat hippocampal slices and in organotypic hippocampal slice cultures. In control Krebs, glutamate and non-N-methyl-D-aspartate (NMDA) agonist-induced responses were enhanced by zinc (25-300 microM). However, under conditions favouring NMDA receptor activation, zinc inhibited glutamate- and NMDA-induced responses. Metabotropic glutamate receptor-mediated responses activated in cultured slices by 1-amino-cyclopentane-1,3-dicarboxylate (1S,3R-ACPD) or by quisqualate, were reversibly inhibited by zinc (200 microM). These results indicate that zinc can inhibit responses induced by activation of metabotropic glutamate receptors and reaffirm that zinc has a differential effect on NMDA and non-NMDA receptors.

Animals↗
PubMed1993

Cloning and functional expression of a brain G-protein-coupled ATP receptor.

A cDNA encoding a novel member of the G-protein-coupled receptor (GCR) superfamily, an ATP receptor, has been isolated from an embryonic chick whole brain cDNA library by hybridization screening. The encoded protein has a sequence of 362 amino acids (41 kDa) and shares no more than 27% amino acid identity with any known GCR. When expressed as a complementary RNA (cRNA) in Xenopus oocytes a slowly-developing inward current was observed in response to application of ATP. The pharmacology of this expressed protein defines it as a P2Y purinoceptor.

Adenosine Triphosphate↗
PubMed1993

Giant GABAB-mediated synaptic potentials induced by zinc in the rat hippocampus: paradoxical effects of zinc on the GABAB receptor.

The interaction of zinc with pre- and postsynaptic GABAB receptors was studied in adult rat hippocampal slices using intracellular recording in CA1 and CA3 pyramidal neurons. Zinc (50-300 microM) antagonized baclofen responses with a variable potency, whereas CGP-35348 (100 microM) or barium (300 microM) produced a more substantial and consistent inhibition. Zinc also induced giant GABAA-mediated depolarizing potentials (GDP) in these neurons. After blocking GABAA and excitatory synaptic transmission, monosynaptic hyperpolarizing inhibitory postsynaptic potentials (IPSP) mediated by GABAB receptors (IPSPB) were inhibited by CGP-35348 or barium; however, zinc increased the latency and prolonged the duration of the IPSPB and also induced the appearance of spontaneous giant GABAB-mediated hyperpolarizing potentials (GHP). In some cells, IPSPBs in zinc exhibited a multiphasic appearance. The early component was partially inhibited by 300 microM zinc and was followed by a late GHP. CGP-35348 at 100 microM inhibited the early monosynaptic IPSPB but not the GHP; however, at 300 microM both components were blocked. Paired-pulse inhibition of the IPSPB was used to assess the effect of zinc on presynaptic GABAB receptors. Neither the zinc-chelating agent CP94 (400 microM) nor zinc affected this phenomenon. CGP-35348, barium and polyvalent cations, such as cadmium, copper, cobalt, manganese, iron and aluminum, failed to induce giant potentials in hippocampal neurons. It is concluded that zinc is apparently unique in synchronizing the release of GABA to produce GDPs and GHPs.

Animals↗
PubMed1993

Interplay between expressed non-NMDA receptors and endogenous calcium-activated chloride currents in Xenopus laevis oocytes.

Current-voltage (I-V) relationships of glutamate receptors activated by the non-NMDA receptor agonist, kainate (KA), were determined in Xenopus laevis oocytes injected with either calf or chick brain mRNA. In most injected oocytes (n = 44; 84%), I-V plots to 80 microM KA exhibited inward rectification; however, in some oocytes (n = 7; 16%), the KA-evoked membrane current showed only slight outward rectification. A comparison between the resting membrane properties of injected oocytes with the KA-evoked currents revealed that weak outward rectification was often observed in oocytes possessing predominant voltage-activated calcium-dependent chloride currents (ICl(Ca)). In these oocytes, replacement of extracellular calcium (2 mM Ca2+) with magnesium ions (Mg2+), or the addition of the chloride channel blocker, anthracene-9-carboxylic acid (500 microM to 1 mM A9C), reduced ICl(Ca) and also altered the nature of the KA I-V plot revealing inward rectification. It is proposed that the responses mediated by expressed non-NMDA receptors may be influenced by the activation of endogenous calcium-dependent membrane currents in Xenopus laevis oocytes.

Animals↗
PubMed1993

Thiocyanate ions selectively antagonize AMPA-evoked responses in Xenopus laevis oocytes microinjected with rat brain mRNA.

1 Responses to kainate (KA), willardiine and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) were recorded from rat brain mRNA-injected Xenopus laevis oocytes by use of a two-electrode voltage clamp. 2 Thiocyanate (SCN-; 50 microM-4 mM) ions reversibly and selectively inhibited the membrane current responses to AMPA in a non-competitive manner without affecting KA or willardiine-induced responses. 3 The inhibition of AMPA-induced responses by SCN- was dependent on the SCN- concentration with an estimated IC50 of 1 mM. The antagonism was not dependent on the AMPA concentration. 4 The response to a high concentration of AMPA (100-200 microM) exhibited a peak inward current which declined to a steady-state. SCN- inhibited the steady-state current more than the peak response. The inhibition was unaffected by prior incubation with concanavalin-A (Con-A; 10 microM). 5 Responses to KA were antagonized by AMPA in a competitive manner, suggesting that both agonists may activate a common receptor-channel complex. This interaction between two non-NMDA agonists was not affected by the SCN(-)-induced inhibition of the AMPA response. 6 AMPA-induced responses recorded from large cultured cerebellar neurones by whole-cell recording were also inhibited by SCN- in a non-competitive manner. The AMPA-induced peak current was less affected than the steady-state response. 7 We conclude that SCN- can inhibit the response to AMPA in expressed non-NMDA receptors in Xenopus oocytes and also in native receptors in cultured cerebellar neurones. One possible mechanism of action for SCN- inhibition of responses to AMPA may involve a Con-A-insensitive, non-NMDA receptor-mediated desensitization.

Alanine↗
PubMed1993

Properties of GABA-mediated synaptic potentials induced by zinc in adult rat hippocampal pyramidal neurones.

1. Intracellular recording techniques were used to study the actions of the transition ion, zinc, on CA1 and CA3 pyramidal neurones in adult rat hippocampal slices. 2. Zinc (300 microM) hyperpolarized pyramidal neurones, increased the membrane excitability and also induced periodic, spontaneous giant depolarizing potentials associated with a conductance increase mechanism. 3. The occurrence of spontaneous giant depolarizations was dependent on the zinc concentration (10 microM-1 mM) with an apparent dissociation constant of 98 microM. The frequency of zinc-induced depolarizations was unaffected by the membrane potential from -50 to -100 mV. 4. Stimulation of the Schaffer collaterals or mossy fibre pathways evoked an excitatory and inhibitory synaptic potential complex. In the presence of zinc, nerve fibre stimulation evoked, in an all-or-none fashion, a giant depolarizing potential with an increased membrane conductance. Both spontaneous and evoked depolarizations were inhibited by 1 microM tetrodotoxin. 5. Evoked giant depolarizations were labile with too frequent stimulation resulting in a failure of generation. A minimum time of 140 s was required between stimuli to ensure successive giant depolarizations. 6. Spontaneous and evoked zinc-induced depolarizing potentials were inhibited by bicuculline (10 microM) or picrotoxin (40 microM) and enhanced by pentobarbitone (100 microM) or flurazepam (10 microM), suggesting that these potentials are mediated by activation of gamma-aminobutyric acidA (GABAA) receptors. 7. Ionophoretic application of GABA produced biphasic responses at -60 mV membrane potential. The reversal potentials for the depolarizing and hyperpolarizing GABA responses were -56 +/- 5 and -66 +/- 8 mV respectively. The giant depolarizations induced by zinc reversed at -57 +/- 4 mV. This suggests a dendritic location for the generation of these potentials. 8. Excitatory amino acid antagonists, 2-amino-5-phosphonovalerate (APV, 40 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) did not affect the amplitude but slightly reduced the frequency of the giant depolarizations. 9. It is concluded that zinc induces a synchronized release of GABA, quite independent of intact excitatory synaptic transmission, which acts on GABAA receptors producing large depolarizing synaptic potentials. This increased level of GABA release may be of physiological and pathological importance since zinc is a naturally occurring metal ion endogenous to the central nervous system.

Action Potentials↗
PubMed1992

gamma-Hydroxybutyrate depresses monosynaptic excitatory and inhibitory postsynaptic potentials in rat hippocampal slices.

The action of gamma-hydroxybutyrate was studied on pre- and postsynaptic GABA(B) receptors in rat hippocampal neurones in vitro using intracellular recording. gamma-Hydroxybutyrate (1-10 mM) caused a 4-8 mV hyperpolarization of CA1 cells and a 20-80% decrease in monosynaptic excitatory and inhibitory postsynaptic potentials in a concentration-dependent manner. These actions were reversibly inhibited by a novel and selective GABA(B) antagonist, CGP 36742 (20-500 microM) suggesting that gamma-hydroxybutyrate can activate presynaptic as well as postsynaptic GABA(B) receptors.

Animals↗
PubMed1992

Functional modulation of GABAA receptors by cAMP-dependent protein phosphorylation.

gamma-Aminobutyric acidA (GABAA) receptors are ligand-gated ion channels that mediate inhibitory synaptic transmission in the central nervous system. The role of protein phosphorylation in the modulation of GABAA receptor function was examined with cells transiently transfected with GABAA receptor subunits. GABAA receptors consisting of the alpha 1 and beta 1 or the alpha 1, beta 1, and gamma 2 subunits were directly phosphorylated on the beta 1 subunit by adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase (PKA). The phosphorylation decreased the amplitude of the GABA response of both receptor types and the extent of rapid desensitization of the GABAA receptor that consisted of the alpha 1 and beta 1 subunits. Site-specific mutagenesis of the serine residue phosphorylated by PKA completely eliminated the PKA phosphorylation and modulation of the GABAA receptor. In primary embryonic rat neuronal cell cultures, a similar regulation of GABAA receptors by PKA was observed. These results demonstrate that the GABAA receptor is directly modulated by protein phosphorylation and suggest that neurotransmitters or neuropeptides that regulate intracellular cAMP levels may modulate the responses of neurons to GABA and consequently have profound effects on synaptic excitability.

Animals↗
PubMed1992

Gamma-hydroxybutyrate hyperpolarizes hippocampal neurones by activating GABAB receptors.

gamma-Hydroxybutyrate, a naturally occurring substance present in the mammalian central nervous system caused a dose-dependent (0.25-10 mM) hyperpolarization and small membrane conductance increase when applied to hippocampal CA1 pyramidal neurones in vitro. This action was reversibly inhibited by the GABAB antagonist, CGP 35348 (20-100 microM) and divalent cations, Zn2+ and Ba2+, but not by the GABAA antagonist bicuculline (50 microM). These results suggest that GABAB receptors may mediate the actions of gamma-hydroxybutyrate.

Animals↗
PubMed1992

A novel modulatory binding site for zinc on the GABAA receptor complex in cultured rat neurones.

1. The properties of gamma-aminobutyric acidA (GABAA) receptor-ion channel complexes and the interaction with the transition metal zinc, were studied on rat sympathetic and cerebellar neurones in dissociated culture using patch clamp recording techniques. 2. The antagonism of GABA-induced membrane currents by zinc on sympathetic neurones was subject to developmental influence. Using embryonic sympathetic neurones acutely cultured for 24-72 h, GABA responses were more depressed by zinc when compared to responses evoked on adult neurones cultured for the same period. For neurones developing in vivo, the percentage inhibition of GABA responses produced by zinc in embryonic neurones was estimated to decline by 50% after 48.2 days following birth. 3. Embryonic sympathetic neurones maintained in culture for prolonged periods (40-50 days in vitro, DIV) became less sensitive to zinc when compared to neurones cultured for shorter periods (10-20 DIV). The decrease in the zinc inhibition for neurones maintained in vitro proceeded at an apparent rate of 0.55% per day. 4. Activation of the GABA receptor by muscimol (0.2-2 microM) was also antagonized by zinc (50-100 microM). 5. Lowering the pH of the perfusing Krebs solution did not affect the inhibition of GABA responses by zinc on sympathetic neurones. 6. Modulation of the GABAA receptor by some benzodiazepines, a barbiturate, a steroid based on pregnanolone, or antagonists bicuculline and picrotoxinin, did not interfere with the antagonism exerted by zinc on sympathetic neurones. A novel binding site for zinc on the GABAA receptor is proposed. 7. Analysis of the GABA-activated current noise on sympathetic neurones revealed two kinetic components to the power spectra requiring a double Lorentzian fit. The time constant describing the fast component (tau 2, 2.1 ms) was unaffected by zinc, whereas the slow component time constant (tau 1, 21.7 ms) was slightly reduced to 17.1 ms. 8. The apparent single-channel conductance for GABA-activated ion channels was determined from the power spectra (gamma s = 22.7 pS) and also from the relationship between the mean GABA-induced inward current and the variance of the current (gamma v = 24 pS). Zinc (25-100 microM) did not affect the single-channel conductance. 9. Single GABA-activated ion channels were recorded from outside-out patches taken from the soma of large cerebellar neurones. Single GABA channels were capable of activation to multiple current amplitudes which were assessed into the following conductance levels: 8, 18, 23, 29 and 34 pS.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

A physiological role for endogenous zinc in rat hippocampal synaptic neurotransmission.

The mammalian central nervous system (CNS) contains an abundance of the transition metal zinc, which is highly localized in the neuronal parenchyma. Zinc is actively taken up and stored in synaptic vesicles in nerve terminals, and stimulation of nerve fibre tracts that contain large amounts of zinc, such as the hippocampal mossy fibre system, can induce its release, suggesting that it may act as a neuromodulator. The known interaction of zinc with the major excitatory and inhibitory amino-acid neurotransmitter receptors in the CNS supports this notion. That zinc has a role in CNS synaptic transmission, however, has so far not been shown. Here we report a physiological role for zinc in the young rat hippocampus (postnatal, P3-P14 days). Our results indicate that naturally occurring spontaneous giant depolarizing synaptic potentials (GDPs) in young CA3 pyramidal neurones, mediated by the release of GABA (gamma-aminobutyric acid), are induced by endogenously released zinc. These synaptic potentials are inhibited by specific zinc-chelating agents. GDPs are apparently generated by an inhibitory action of zinc on both pre- and postsynaptic GABAB receptors in the hippocampus. Our study implies that zinc modulates synaptic transmission in the immature hippocampus, a finding that may have implications for understanding benign postnatal seizures in young children suffering with acute zinc deficiency.

Action Potentials↗
PubMed1991

Interaction of 5-bromowillardiine with non-NMDA receptors expressed in Xenopus laevis oocytes injected with chick brain mRNA.

Excitatory amino acid (EAA) receptors were studied in Xenopus laevis oocytes microinjected with 1-day-old chick brain mRNA using a two-electrode voltage clamp technique. EAA agonists could be differentially characterised according to their current/voltage (I/V) relationships; I/V plots to kainate (KA) and domoate (Dom) inwardly rectified at positive membrane potentials whereas quisqualate (QA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and L-glutamate (L-Glu) produced linear I/V plots. Interestingly, the non-NMDA (N-methyl-D-aspartate) agonist, 5-bromowillardiine (BrW), displayed properties similar to KA and Dom. It is proposed that BrW acts as a KA-like agonist on chick EAA receptors expressed in Xenopus oocytes.

Alanine↗
PubMed1991

GABAA receptors are differentially sensitive to zinc: dependence on subunit composition.

GABAA receptors with different subunit composition, were expressed in kidney cells and studied by whole cell recording. Expressed GABAA receptors were differentially sensitive to inhibition by zinc; receptors which lacked the gamma subunit were inhibited by zinc. Embryonic neurons also exhibited zinc-sensitive GABA responses, in contrast to adult neurones. This developmentally-sensitive aspect of GABAA receptor pharmacology may be partly dependent on expression of the gamma subunit.

Animals↗