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R Grantyn

Publications and source records attributed to R Grantyn.

At least 37 records · Page 2Linked to original sources
PubMed1994

GABA-activated chloride currents of postnatal mouse retinal ganglion cells are blocked by acetylcholine and acetylcarnitine: how specific are ion channels in immature neurons?

The goal of this study was to clarify pharmacological properties of GABAA receptors in cells of the mouse retinal ganglion cell layer in situ. Spontaneous synaptic currents and responses to exogenous GABA were recorded from individual neurons in retinal whole mounts (postnatal days 1-3) or retinal stripe preparations (postnatal days 4-6). Drugs were applied by a fast local superfusion system. Current responses were measured with the patch-clamp technique in the whole-cell configuration. All cells responded to exogenous GABA (average EC50 and Hill coefficient: 16.7 microM and 0.95 respectively) and generated GABAergic synaptic currents in response to elevated KCl. GABA-induced currents of retinal ganglion cells were blocked by bicuculline, picrotoxin and Zn2+, as well as strychnine, and increased by pentobarbital, clonazepam and 3 alpha-hydroxy-5 alpha-pregnan-20-one. In some retinal ganglion cells GABA caused an increase in the frequency of spontaneous synaptic currents, which points to a partially depolarizing action of this traditionally inhibitory neurotransmitter in the neural retina. Our major observation is that acetylcholine and acetylcarnitine blocked or reduced GABAergic inhibitory postsynaptic currents and responses to exogenous GABA. This effect was seen in only a fraction of retinal ganglion cells and occurred in both the undesensitized and the desensitized state of the GABAA receptor. The block was voltage-independent and persisted during coapplication with the nicotinic and muscarinic acetylcholine receptor antagonists D-tubocurarine and atropine. In contrast to GABA-activated Cl- currents, glycine-activated Cl- currents remained unaffected by acetylcholine and acetylcarnitine.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcarnitine↗
PubMed1994

Ligand- and voltage-gated ion channels are expressed by embryonic mouse retinal neurones.

The present study was intended to investigate whether voltage- and ligand-activated ion channels are expressed during prenatal development by neurones located in the ganglion cell layer of the mammalian retina. Whole cell patch clamp recordings from presumed mouse retinal ganglion cells revealed the expression of Na+, K+ and Ca2+ channels, predominantly of the low-voltage-activated type. Using local application of transmitter substances we further demonstrated that these cells are endowed with glutamate receptors of the N-methyl-D-aspartate (NMDA) and non-NMDA type as well as nicotinic acetylcholine, gamma-amino-butyric acid (GABA)A and glycine receptors. Voltage-gated conductances probably underlie spontaneous action potential generation by embryonic ganglion cells. The early expression of transmitter-gated ion channels indicates important functions of these channels in cell differentiation processes.

Action Potentials↗
PubMed1994

Separation of calcium currents in retinal ganglion cells from postnatal rat.

A culture system of the postnatal rat retina was established to investigate Ca2+ currents and synaptic transmission in identified neurons. Methods are described that allowed us to select retinal ganglion neurons (RGNs) in short term cultures (up to 48 h in vitro) and in long-term cultures (3 to 21 days in vitro). The specific aim of the present study was to identify channel specific components in whole-cell Ca2+ currents of RGNs and to clarify the potential use of the lanthanide Gd3+ as a selective Ca2+ channel blocker. About one third of freshly dissociated RGNs generated both low voltage activated Ca2+ currents (ICa(LVA)) and high voltage activated Ca2+ currents (ICa(HVA)). The remaining 2/3 or RGNs in short term culture and most RGNs in long-term culture displayed only ICa(HVA). The latter comprised at least three different components that were functionally rather similar, but could be separated pharmacologically. A significant portion (about 40%) of ICa(HVA) was irreversibly blocked by the N channel antagonist omega-CgTx (5 microM). The L channel antagonist nifedipine (10 microM) eliminated about 25% of ICa(HVA). Thus, about 1/3 of the HVA Ca2+ or Ba2+ current remained unaffected by either omega-CgTx or nifedipine. omega-AgaTx (200 nM) completely failed to block HVA Ca2+ or Ba2+ currents in RGNs. Gd3+ exerted contrasting actions on LVA and HVA Ca2+ currents. While ICa(LVA) consistently increased in the presence of Gd3+ (0.32-3.2 microM), ICa(HVA) always decreased, especially when using higher concentrations of Gd3+ (10-32 microM). The blocking action of Gd3+ was not restricted to the omega-CgTx-sensitive HVA current component, but also concerned omega-CgTx- and nifedipine-resistant components. The decay of Ca2+ currents was accelerated in the presence of Gd3+. Even in RGNs lacking ICa(LVA), application of 3.2 microM Gd3+ significantly reduced the time constant of decay from an average of 64 ms to 36 ms (voltage steps from -90 to 0 mV; 10 mM [Ca2+]o; 26 degrees C). This is in contrast to what had to be expected if an N-type HVA current component was selectively suppressed by Gd3+.Gd3+ diminished glutamatergic spontaneous synaptic activity in retinal cultures tested during the 3rd week in vitro. Both frequency and amplitude were reduced. Occasionally, the application was followed by a rebound increase of EPSC frequency. A stimulatory effect during application of Gd3+ has never been observed. These experiments indicate that RGNs express at least 4 different types of Ca2+ currents, that resemble in some aspects T, N and L channel currents.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Potentiating and depressant effects of metabotropic glutamate receptor agonists on high-voltage-activated calcium currents in cultured retinal ganglion neurons from postnatal mice.

This study was aimed at clarifying the role of metabotropic glutamate receptors (mGluRs) in the regulation of intracellular Ca2+ concentration ([Ca2+]i in postnatal mouse retinal ganglion neurons (RGNs). RGNs were maintained for 1-2 weeks in vitro by adding brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF) to the culture medium. In order to select these cells for electrophysiological measurements, RGNs were vitally labelled with an antibody against Thy-1.2. Voltage-activated Ca2+ currents [ICa(V)] were recorded with patch electrodes in the whole-cell configuration. It was found that racemic +/--1-amino-cyclopentane-trans-1,3-dicarboxylic acid (t-ACPD) or its active enantiomer 1S,3R-ACPD rapidly and reversibly either enhanced or depressed ICa(V). Quisqualate (QA), L-2-amino-4-phosphonobutyrate (L-AP4) and the endogenous transmitter glutamate induced similar effects when ionotropic glutamate receptors were blocked with D-2-amino-5-phosphonovalerate (D-APV) and 6,7-dinitroquinoxaline-2,3-dione (DNQX). omega-Conotoxin GVIA (omega-CgTx GVIA), but not nifedipine prevented modulation of ICa(V) by mGluR agonists. The depression of ICa(V) by t-ACPD was irreversible when cells were dialysed with guanosine-5'-O-(3-thiotriphosphate) (GTP[gamma-S]). Ratio measurements of fura-2 fluorescence in Thy-1+ cells showed that neither t-ACPD, QA nor L-AP4 affected [Ca2+]i by liberation of Ca2+ from intracellular stores. Our results suggest that cultured RGNs express mGluRs. These receptors cannot induce Ca2+ release from intracellular stores but regulate [Ca2+]i by a fast and reversible, G-protein-mediated action on a subpopulation of voltage-activated Ca2+ channels.

Alanine↗
PubMed1994

In vitro identification of retinal ganglion cells in culture without the need of dye labeling.

We here describe a method for the identification of a distinct neuronal phenotype at all stages of development in culture without the need of any staining procedure. Based purely on a size criterion we can rapidly select vital retinal ganglion cells (RGCs) for further studies out of a mixed culture of rat retinal cells. In order to establish a size criterion for retinal cells of various age, RGCs were first labeled immunocytochemically with antibody against the ganglion cell-specific surface glycoprotein Thy-1. Soma diameters were then determined for labeled and unlabeled cells between embryonic day 16 (E16) and postnatal day 90 (P90). Unlabeled neurons of all ages had soma diameters between 3.6 microns and 12 microns (mean diameter: 6.3 microns). In contrast, soma diameters of RGCs ranged from 8.4 microns to 28 microns and the number of RGCs with large soma diameters increased with age. Thus, in a mixed retinal cell culture only RGCs are larger than 12 microns and can be selected solely based on their size. The validity of the size criterion during the whole period of retinal cell differentiation offers the possibility to study the development of cellular functions and ion channel properties in a distinct type of cell without the risk of artifacts introduced by staining.

Animals↗
PubMed1994

Retinal ganglion neurons express a toxin-resistant developmentally regulated novel type of high-voltage-activated calcium channel.

1. High-voltage-activated Ca2+ currents [ICa(HVA)] were studied in immunolabeled mouse retinal ganglion neurons (RGNs) to elucidate channel-specific components and their developmental changes in vitro. 2. Neurons were dissociated at postnatal day 5. RGNs were selected for electrophysiological measurements by vital labeling with an antibody against Thy-1.2. ICa(HVA) were recorded with patch electrodes in the whole cell configuration at a holding voltage (Vh) of -90 mV. 3. A total of 111 neurons was studied. On average, 13% of ICa(HVA) was reversibly blocked by 10 microM nifedipine, approximately 30% of the compound current displayed an irreversible block by 2.5 microM omega-conotoxin (omega-CTX) GVIA. The remainder current was resistant to both drugs, suggesting that the total ICa(HVA) was a mixture of at least three different components. 4. Developmental analysis revealed a significant increase of the omega-CTX-GVIA/nifedipine-resistant component of ICa(HVA) (31% at day in vitro (DIV) 0-2, 70% at DIV 18-26) mainly at the expense of the omega-CTX-GVIA-sensitive current. No significant change was found in the nifedipine-sensitive component of ICa(HVA). 5. To characterize the Ca2+ current component that was resistant to both omega-CTX-GVIA and nifedipine at Vh -90 mV, three tests were performed. The P channel antagonist omega-agatoxin IVA (omega-Aga-IVA, 200 nM) completely failed to block ICa(HVA) in mouse RGNs. The novel Ca2+ channel blocker omega-CTX-MVIIC (5 microM) decreased the ICa(HVA) remaining after omega-CTX-GVIA treatment by only approximately 10%.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1993

Glutamatergic and GABAergic synaptic currents in ganglion cells from isolated retinae of pigmented rats during postnatal development.

This study was aimed at characterizing the earliest phases of synaptogenesis in the mammalian retina. Spontaneous activity of ganglion cells in the isolated superfused retina was used as an indicator for the functionality of synaptic connections. Retinal ganglion neurons (RGNs) were identified by location of their somata in the ganglion cell layer (GCL) and by their ability to generate large (> 500 pA) voltage-activated sodium currents. Spontaneous spiking was found in many RGNs prior to cell perfusion. Between postnatal day (P) 1 and 18, a total of 195 RGNs was tested for light-induced currents, conductance changes in response to exogenous glutamate (Glu) and gamma-aminobutyric acid (GABA), and depolarizing or hyperpolarizing synaptic activity. The vast majority of the material was derived from RGNs at day P5. Whole-cell ion currents were always sampled at somatic sites, using either conventional or perforated patch whole-cell recordings. On day P5, 5% of tested RGNs (n = 73) were already responsive to light stimulation. A higher percentage of cells (23%, n = 187) generated spontaneous depolarizing currents that were regarded as glutamatergic excitatory postsynaptic currents (EPSCs), since (1) they were blocked by Glu antagonists, (2) they conformed to the Na+/Cs+ equilibrium potential, (3) and they displayed a time course characteristic of glutamatergic EPSCs. The mean EPSC amplitude was 19.0 pA (S.D. 11.83 pA). Amplitude distributions were fitted by multiple Gaussian equations rendering a quantal size of 6.6 to 9.1 pA at a holding voltage (Vh) of -70 mV (driving force about 70 mV). Spontaneous EPSCs were never observed under condition of Ca(2+)-free solutions, but they persisted in the presence of tetrodotoxin. Bath application of quisqualate (500 microM) consistently increased EPSC frequencies. In contrast to the relatively high percentage of RGNs generating spontaneous EPSCs, very few RGNs at P5 (3%, n = 187) displayed inhibitory postsynaptic currents (IPSCs), although by that time all tested RGNs (n = 14) were responsive to both exogenous Glu and GABA. These results indicate that in the postnatal rat retina development of excitatory synapses precedes the maturation of inhibitory afferents. Excitatory inputs to RGNs were to some extent functional before the animals opened their eyes. Glutamatergic synaptic activity may, thus, play an important role in shaping visual connections in the absence of visual experience.

Animals↗
PubMed1993

Rat retinal ganglion cells express Ca(2+)-permeable non-NMDA glutamate receptors during the period of histogenetic cell death.

Local application of glutamate agonists to retinal ganglion cells (RGNs) was performed in retinae isolated from pigmented rats aged between 3 and 8 days postnatally. A vast majority of RGNs displayed current responses to glutamate (Glu), N-methyl-D-aspartate (NMDA), quisqualate (QA), alpha-amino-2,3-dihydro-5-methyl-3-oxo-4-isoazolepropanoic acid (AMPA), kainate (KA) and domoate (DA). In Na(+)-free extracellular solution with elevated Ca2+, non-NMDA agonists elicited large (up to 200 pA) inward currents that were completely blocked by 6,7-dinitroquinoxaline-2,3-dione (DNQX) and Cd2+. MK-801 also induced a partial block of cationic currents in Na(+)-free saline. In standard salt solutions, current-voltage relationships of Glu-R-mediated currents were often inwardly rectifying in the presence of D-aminophosphonovalerat (D-APV), as is typical of Ca(2+)-permeable non-NMDA receptors. The presence of inward rectification in the current voltage relationship was always associated with a high value of the cationic permeability ratio PCa2+/PCs+ (> 0.8). However, in about half of the investigated RGNs no inward rectification was observed under standard recording conditions. Our results lead to the suggestion that expression of Ca(2+)-permeable Glu receptor subunits may contribute to regulation of cell numbers in the postnatal retina.

Amino Acids↗
PubMed1992

Development of GABAergic connections in vitro: increasing efficacy of synaptic transmission is not accompanied by changes in miniature currents.

Development of inhibitory synaptic transmission was studied using a dissociated cell culture from the superior colliculus of neonatal rat. Patch-clamp recordings in the whole-cell configuration were performed to measure evoked (single-cell-activated) inhibitory postsynaptic currents (IPSCs), miniature IPSCs and current responses to maximal concentrations of exogenous gamma-aminobutyric acid (GABA). Over a period of 3 weeks in vitro (DIV3-24), the fraction of synaptically coupled neurons raised from 0% to 76%. Evoked IPSCs were first observed at DIV5. They had an average amplitude of 33.9 pA during the first week (n = 13) and 129.7 pA during the fourth week (n = 48). This increase by a factor of 3.8 represents a significant rise in the efficacy of GABAergic transmission during in vitro development. However, no developmental change has been observed in the average amplitudes of miniature somatic IPSCs. The latter remained at an average level of about 9 pA (symmetrical chloride concentration and a driving force of 68 mV). No increase was found also in whole-cell current densities induced by saturating concentrations of exogenous GABA. Our results suggest that under the given conditions, synapse maturation was primarily the result of presynaptic sprouting. This conclusion is further supported by bouton counts in immunostained collicular cultures, where the number of axosomatic and axodendritic GABAergic contacts per neuron increased from 0.54 and 0.37, respectively, at DIV3, to 13.84 and greater than 23.1, at DIV24. The overall density of GABAergic neurons decreased during this period from about 41,000/cm2 to 15,600 cm2, indicating that a growing number of contacts is formed by a declining number of presynaptic neurons.

Animals↗
PubMed1992

Unitary, quantal and miniature GABA-activated synaptic chloride currents in cultured neurons from the rat superior colliculus.

The aim of this study was to identify the conductance change induced by one quantum of gamma-aminobutyric acid from axonal release sites on cultured superior colliculus neurons. Unitary (single cell-activated) inhibitory postsynaptic currents and spontaneous synaptic activity were recorded with patch clamp techniques in the whole cell configuration while superfusing the entire neuron with normal saline. Miniature inhibitory postsynaptic currents were recorded in the presence of tetrodotoxin and in reduced [Ca2+]o/[Mg2+]o. In addition, the membrane area contributing to synaptic activity was limited to a narrow window of 50 microns. Smaller neurons were chosen for recording to render a standard deviation of the "instrumental" noise of less than 1.5 pA at a holding voltage of -80 mV. After two weeks in vitro, the percentage of synaptically connected tectal neurons exceeded 50%. At holding voltages of -80 mV (Cl- equilibrium potential -12 mV) minimal amplitudes of unitary inhibitory postsynaptic currents were as low as 7-10 pA, while maximal amplitudes exceeded 500 pA. The mean time to peak and time constant of decay were 3.0 and 34.4 ms, respectively (n = 31). Fluctuating unitary inhibitory postsynaptic currents were deemed to be compound postsynaptic responses. Multiple Gaussian equations could be fitted to the amplitude histograms of unitary postsynaptic currents. This procedure rendered a quantal size between 5.0 and 10.9 pA (mean 7.1 pA; S.D. 1.78 pA) in five neurons from mature cultures. The amplitudes of statistically determined quantal inhibitory postsynaptic currents were slightly smaller than the independent estimate from somatic miniature inhibitory postsynaptic currents. The latter had a mean amplitude of 9.1 pA (S.D. 3.3 pA, n = 23), a mean time to peak of 1.65 ms (n = 9), and a mean time constant of decay of 16.2 ms (n = 9). Single channel recording from outside-out patches showed three to four main conductance states ranging from 9 to 22 pS. Single channel closures at the 21-24 pS level were occasionally observed during relaxation of miniature currents. The small size of whole cell quantal inhibitory postsynaptic currents and somatic miniature currents indicates that one GABA quantum opened only 5-15 single Cl- channels.

Animals↗
PubMed1990

Is GABA release modulated by presynaptic excitatory amino acid receptors?

The purely GABAergic nature of spontaneous synaptic activity in cultures from the neonatal rat superior colliculus (SC) is of great advantage in investigations aimed at characterizing presynaptic factors regulating GABAergic synaptic transmission. Using SC-derived cultures it was confirmed that excitatory amino acids (EAA) can induce a marked increase in the frequency of spontaneous synaptic Cl- currents (ICl(GABA)SYN). However, this tetrodotoxin-resistant facilitation of Ca2(+)-dependent GABA release required application of EEA to several neurons (multiple cell superfusion). In contrast, no frequency increase of Icl(GABA)SYN was seen with restricted access of EAA to only one neuron and the presynaptic axonal terminals (single cell superfusion). It is therefore concluded that the strong facilitatory effect of glutamate (Glu) and kainate (KA) on GABAergic synaptic activity, as observed under the condition of multiple cell superfusion, is mediated via somatodendritic excitatory amino acid receptors (EAARs).

Action Potentials↗
PubMed1990

Development of GABAergic synaptic connections in vivo and in cultures from the rat superior colliculus.

Synaptic activity in the superficial (i.e. visual) layer of the superior colliculus was investigated with intracellular microelectrodes using a preparation of the isolated superfused tectum from neonatal rat. It was found that by postnatal day 9 (i.e. before eye opening) the majority of neurons in the superficial gray layer (SGS, stratum griseum superficiale) were already capable of generating Cl(-)-dependent inhibitory postsynaptic potentials (IPSPs) in response to intracollicular stimulation. Properties and development of GABAergic synaptic connections were further characterized in a dissociated cell culture from the SGS. The cultures were prepared from E21 rat embryos and studied between 1 and 38 days in vitro (DIV). gamma-[3H]aminobutyric acid ([3H]GABA) uptake served to identify GABAergic neurons and to estimate their relative density. Axon terminals were labeled by indirect immunostaining for glutamic acid decarboxylase (GAD) and examined with light (LM) and electron microscopy (EM). Responsiveness to exogenous and endogenous GABA was investigated by recording ionic currents with patch clamp techniques. [3H]GABA uptake-positive neurons constituted about 40% of the whole cellular population dissociated from the SGS of E21 rats. After 2 weeks in culture, [3H]GABA uptake was observed in 45-60% of the cells with neuronal features. The relative number of GAD-immunoreactive neuronal perikarya ranged from 28 to 39%, after 2 weeks in vitro. Responsiveness to exogenous GABA was found in all freshly plated neurons. Release of GABA could be demonstrated after 2 DIV by recording spontaneous bicuculline-sensitive Cl- currents. These currents had the characteristics of GABAA receptor-mediated synaptic currents. However, even as late as DIV 6, very few vesicle-containing axonal terminals apposing postsynaptic specializations were revealed with EM. GAD-labeled puncta became clearly visible only after DIV 10-12. Between DIV 14 and 21, the intensity of immunostaining and the density of GAD-labeled synaptic contacts increased, reaching a maximum around DIV 28. GAD-positive puncta covered both neurons and non-neuronal cells. At the level of EM, GAD-positive terminals were shown to establish synaptic contacts with neuronal somata and processes, forming in the majority of cases (22 out of 32 stained terminals) symmetrical contacts. It is concluded that in the SGS of the rat superior colliculus GABAergic neurons and GABAA receptors are present before birth. In dissociated cell cultures ionic currents can be generated in response to endogenous GABA before axonal terminals of GABAergic neurons fully mature. Finally, our experiments show that visual activity is not a prerequisite for the formation of GABAergic synapses between neurons of the SGS.

Action Potentials↗
PubMed1989

Expression of depolarizing voltage- and transmitter-activated currents in neuronal precursor cells from the rat brain is preceded by a proton-activated sodium current.

The early expression of amiloride-sensitive proton-activated sodium currents (INa(H] was demonstrated using the giga-seal whole-cell voltage clamp technique in cells from the primordial tectum of E12 rat embryos. Less than 10% of these cells stained for tetanus toxin receptors after 2 h in vitro. However, after 10 h in vitro all cells with neuronal geometry were tetanus toxin-positive and capable of generating voltage-activated Na currents (INa(V] and high-voltage activated Ca2+-currents (ICa(HV]. INa(H) was expressed roughly in parallel with INa(V) and ICa(HV), but exceeded the former currents in amplitude by 50-100 times, reaching 600 pA and more. In 25% of the cells tested within the first 5 h in vitro INa(H) was, in fact, the only cationic inward current resolved. Responses to quisqualate and kainate appeared only after 3 days in vitro, and responses to N-methyl-D-aspartate/glycine were seen only after 4 days in vitro. These results suggest that the channels carrying INa(H) are present at the earliest stages of neuronal development.

Animals↗
PubMed1989

Glial cells of the oligodendrocyte lineage express proton-activated Na+ channels.

Neurons and oligodendrocytes, but not type I astrocytes and Schwann cells, generate large Na+ currents in response to a step increase of [H+]. Proton-activated Na+ channels are the first cationic channels expressed in neuronal precursor cells from the mammalian brain. Glial precursor cells cultured from mouse brain are also capable of generating Na+ currents in response to step acidification (INa(H]. With further development along the oligodendrocyte lineage, this property is retained, whereas voltage-activated Na+ and K+ currents disappear. Comparing INa(H) of oligodendrocytes with INa(H) of their precursor cells did not reveal a difference in current amplitude, suggesting a higher density of INa(H) channels on the (smaller) precursor cells. The properties of INa(H) in glial precursor cells and oligodendrocytes are similar to those of neurons, with respect to activation conditions, time course, and the effect of extracellular Ca2+ concentrations. The results are consistent with previous observations which showed that oligodendrocytes partially preserve their chemically activated, but completely lose their voltage-activated, ion channels.

Animals↗
PubMed1989

Separation of quisqualate- and kainate-selective glutamate receptors in cultured neurons from the rat superior colliculus.

The aim of the present study was to identify and characterize the receptors and ionic channels mediating the compound response of tectal neurons to exogenous L-glutamate (Glu). Particular attention was paid to the question of whether separate receptors and channels exist for quisqualate (QA) and kainate (KA) and, if so, whether binding to one of these receptors would modify the response elicited through the other. Neurons were dissociated from the superficial gray layer of the superior colliculus from E21 or P1 rats. Between days 14 and 21 in vitro, responsiveness of tectal neurons to Glu and related substances was tested by recording the whole-cell currents induced by rapid superfusion with drug-containing salt solutions. Our experiments showed that tectal neurons express at least 3 distinct types of receptors for acidic amino acids. KA-activated currents (I(KA)) differ from QA-activated currents (I(QA)) in their dose-response characteristics, desensitization patterns, selective blockade with kynurenic acid and suppression by elevated [Ca2+]o, I(KA), but not I(QA), is significantly reduced by low levels of [Cl-]o, and the [Cl-]o-dependent shift of the reversal potential for I(KA) suggests that KA promotes a conductance decrease for Cl-. Such an effect has been ascribed to APB-receptors, but L-2-amino-4-phosphonobutyrate (APB) itself failed to induce current responses in tectal neurons. KA was without effect when administered together, and in equimolar concentrations, with QA. The block of I(KA) was, however, surmounted by applying KA at considerably higher concentrations. It is concluded that QA acts as a low-affinity competitive antagonist at the KA site and as a high-affinity agonist at its own receptor. The response to the endogenous ligand Glu reflects properties of all receptors. QA and KA receptors account for 20-30% (QA) and 49-82% (KA) of the compound current elicited with 100 microM Glu. These results indicate that binding of Glu does not, in contrast to QA, produce any significant suppression of the KA-receptor-mediated current component.

Animals↗
PubMed1988

Similarity and mutual exclusion of NMDA- and proton-activated transient Na+-currents in rat tectal neurons.

Vertebrate neurons respond to rapid elevation of [H+]o with transient Na+-selective currents (INa(H]. Since INa(H) and voltage-activated Ca2+-currents (ICa(V] are mutually exclusive and similarly affected by inorganic and organic Ca2+-blockers, it has been suggested that such a Na+-permeable state evolves from protonation of Ca2+-channels. We show here that in cultured neurons from embryonic rat superior colliculus N-methyl-D-aspartate (NMDA) provides conditions for generation of a current identical with INa(H), but without the requirement of an increase in free [H+]o. The transient NMDA-activated current (I(NMDA)T) is occluded by INa(H). Its time course is similar to that of INa(H). Both currents are inactivated by long exposure to high [H+]o. I(NMDA)T displays a linear current-voltage (I-V) relationship under conditions which cause a negative slope in the I-V relationship of the persistent NMDA-activated current (I(NMDA)P). This suggests that the biphasic response of tectal neurons to the glutamate-agonist NMDA results from superposition of two different currents.

Animals↗