GABAA receptor stimulation enhances NMDA-induced Ca2+ influx in mouse cerebral cortical neurons in primary culture

doi:10.1016/0169-328X(94)90195-3

Molecular Brain Research

Volume 27, Issue 1, November 1994, Pages 145-151

https://doi.org/10.1016/0169-328X(94)90195-3 Get rights and content

Abstract

The effect of GABA_A receptor stimulation on $N- methyl- d -aspartate(NMDA)-induced$ [⁴⁵Ca²⁺]influx has been examined using primary cultured cerebral cortical neurons. NMDA induced a dose-dependent increase in [⁴⁵Ca²⁺]influx, which was blocked by MK-801 in a dose-dependent manner. GABA_A receptor agonists significantly enhanced the NMDA-induced [⁴⁵Ca²⁺]influx, and this enhancement was dose-dependently inhibited by bicuculline, although picrotoxin and tert-butylbicyclo[2.2.2]phosphoro-thionate (TBPS) exhibited no alterations in this stimulatory action of GABA_A receptor agonists. Blockers of L-type voltage-dependent calcium channels significantly reduced the NMDA-induced [⁴⁵Ca²⁺]influx. The increased [⁴⁵Ca²⁺]influx by both NMDA and GABA_A receptor agonists was also reduced by verapamil and nifedipine. These results suggest that the enhancement of NMDA-induced [⁴⁵Ca²⁺]influx by GABA_A receptor stimulation in immature cerebral cortical neurons may be due to the increased opening of voltage-dependent calcium channel by synergestic actions between NMDA and GABA_A receptors.

References (31)

E. Cherubini et al.
GABA mediated excitation in immature rat CA3 hippocampal neurons
Int. J. Dev. Neurosci.
(1990)
E. Cherubini et al.
GABA: an excitatory transmitter in early postnatal life
Trends Neurosci.
(1991)
D.W. Choi
Glutamate neurotoxicity and diseases on the nervous system
Neuron
(1988)
D. Janigro et al.
Effects of GABA and baclofen on pyramidal cells in the developing rabbit hippocampus: an ‘in vitro’ study
Dev. Brain Res.
(1988)
R.S.G. Jones
Epileptiform events induced by GABA-antagonists in entorhinal cortical cells in vitro are partly mediated by $N- methyl- d -aspartate$ receptors
Brain Res.
(1988)
A.R. Kriegstein et al.
Cellular and synaptic physiology and epileptogenesis of developing rat neocortical neurons in vitro
Dev. Brain Res.
(1987)
K. Kuriyama et al.
Development of γ-aminobutyric acid (GABA)ergic neurons in cerebral cortical neurons in primary culture
Brain Res.
(1987)
O.H. Lowry et al.
Protein measurement with the Folin phenol reagent
J. Biol. Chem.
(1951)
B. Meldrum et al.
Excitatory amino acid neurotoxicity and neurodegenerative disease
Trends Pharmacol. Sci.
(1990)
D. Muller et al.
Developmental changes in synaptic properties in hippocampus of neonate rats
Dev. Brain Res.
(1989)

S. Ohkuma et al.

Muscimol prevents neuronal injury induced by NMDA

Jpn. J. Pharmacol.

(1994)

S. Ohkuma et al.

Development of taurine biosynthesizing system in cerebral cortical neurons in primary culture

Int. J. Dev. Neurosci.

(1986)

S.M. Rothman et al.

Excitotoxicity and the NMDA receptor

Trends Neurosci.

(1987)

W. Sieghart

GABA_A receptor: ligand-gated Cl⁻ ion channels modulated by multiple drug-binding sites

Trends Pharmacol. Sci.

(1992)

R. Yuste et al.

Control of postsynaptic Ca²⁺ influx in developing neocortex by excitatory and inhibitory neurotransmitters

Neuron

(1991)

Cited by (20)

Additive effect of BLA GABA<inf>A</inf> receptor mechanism and (+)-MK-801 on memory retention deficit, an isobologram analysis
2016, Pharmacology Biochemistry and Behavior
Citation Excerpt :
GABAA receptor regulates glutamate release from a population of neocortical glutamatergic neurons via their own presynaptic receptors (Long et al., 2009). For example, GABAA receptor stimulation has been shown to contribute to protective mechanisms involved in neuronal injury induced by NMDA receptor activity (Ohkuma et al., 1994). Numerous evidence supports the notion that GABAA receptor function in the brain is structured via several calcium-regulated mechanisms which are varied in terms of duration, influx source, and intracellular pathway (Gallos et al., 2015; Sametsky et al., 2015).
There is a near correlation between N-methyl-d-aspartate (NMDA) and γ-aminobutyric acid (GABA) receptors in the modulation of learning and memory in the basolateral amygdala (BLA). In this study, we investigated the involvement of GABA_A receptors in the BLA in amnesia induced by (+)-MK-801, a noncompetitive antagonist of NMDA receptors, in male Wistar rats. After guide cannulae were bilaterally placed in the BLA, animals were trained in a step-through type passive avoidance task and then tested 24 h after training to measure memory retrieval and locomotor activity. Post-training intra-BLA microinjection of (+)-MK-801 (0.5 μg/rat) and GABA_A receptor agonists (muscimol at doses 0.05 and 0.1 μg/rat) or antagonist (bicuculline at doses 0.05 and 0.1 μg/rat) decreased step-through latency during retrieval but did not alter locomotor activity. Results also showed that a subthreshold dose of muscimol (0.025 μg/rat) potentiated impairment induced by (+)-MK-801, whereas bicuculline (0.025 μg/rat) restored it. Furthermore, the highest dose of muscimol (0.5 μg/rat) increased locomotor activity induced by (+)-MK-801. Isobologram analysis showed that there was an additive but not synergistic effect between muscimol and (+)-MK-801 on memory retention deficits in the BLA. In conclusion, muscimol and bicuculline decreased retention of memory formation in the BLA, and GABA_A receptors in the BLA may be involved in the additive effect on (+)-MK-801-induced memory retention deficits.
First phase of glucose-stimulated insulin secretion from MIN 6 cells does not always require extracellular calcium influx
2006, Journal of Pharmacological Sciences
To demonstrate an involvement of ATP-sensitive potassium (K_ATP) channel-independent pathways in the first phase of glucose-stimulated insulin secretion (GSIS) from pancreatic β cells, the time course of GSIS from MIN6 cells was analyzed at 30-s sample intervals. GSIS was biphasic with the first phase being observed 120 to 390 s after glucose addition, peaking at 180 s, and with a shoulder at 240 to 330 s. Both 10 µM diazoxide and 3 µM verapamil completely inhibited tolbutamide- or glibenclamide-induced insulin secretion and suppressed the peak of the first phase of GSIS, but did not result in complete suppression. The shoulder following the peak was suppressed by 1 µM dantrolene. The peak, but not shoulder, disappeared under the extracellular Ca²⁺-free condition. A significant amount of insulin secretion remained even in the combined presence of verapamil and dantrolene. The Na⁺ channel blocker tetrodotoxin (30 nM) nearly completely inhibited the first phase release. These results suggest that the first phase of GSIS from MIN6 cells depends on both Ca²⁺-dependent and -independent mechanisms. The former mechanism includes the extracellular Ca²⁺ influx via L-type voltage-dependent calcium channel and intracellular Ca²⁺ release from endoplasmic reticulum via ryanodine receptors, and the latter mechanism involves the pathways associated with Na⁺ channels.
Developmental changes of GABAergic synapses formed between primary cultured cortical neurons
2004, Developmental Brain Research
The characteristics of functional changes of GABAergic synapses between cultured rat cortical neurons were observed by monitoring intracellular calcium level ([Ca²⁺]_in) during development in vitro. After 5 days in vitro (DIV), cultured cortical neurons spontaneously exhibited synchronous oscillatory changes in [Ca²⁺]_in, which were derived from synaptic activity. Exposure to bicuculline, antagonist of γ-aminobutyric acid (GABA)_A receptors, caused a marked decrease in the frequency of [Ca²⁺]_in oscillations at 7–20 DIV. Although the frequency of spontaneous oscillations increased during this culture period, the ratio of the decrease in the frequency following bicuculline treatment did not significantly change. Thereafter, to investigate the detailed morphological changes of GABAergic synapses during development in vitro, the cultured neurons were immunostained with antibodies to glutamic acid decarboxylase (GAD), synaptophysin and GABA_A receptor and were observed under a confocal laser microscope. Most of the GAD-positive puncta colocalized with synaptophysin-positive puncta and were opposed to GABA_A receptor-positive structures. The images of GAD-positive puncta were reconstructed from the confocal three-dimensional data to analyze their number, volume, and surface area. The number of these puncta increased with culture time at 7–20 DIV. Although the volume of individual GAD-positive puncta did not significantly change, the surface area decreased in a time-dependent manner over the culture period. This system that we developed enabled us to investigate in detail the morphological and functional changes of GABAergic synapses during neuronal development.
Ovariectomy has minimal effects on neuroadaptations associated with ethanol dependence in female rats
2000, Neurochemistry International
We previously found gender selective alterations in gene expression for GABA_A and NMDA receptors associated with the development of ethanol dependence. Males and females have a differing hormonal environment, including steroid hormone derivatives (neuroactive steroids) that exert effects at GABA_A and NMDA receptors. Therefore, we explored whether the removal of ovarian steroids would alter gender differences in response to chronic ethanol exposure. We found that ovariectomy reduced ethanol drinking levels by 15%, comparable to earlier observations between intact female and male rats. However, investigation of the effects of chronic ethanol exposure on intact versus ovariectomized female rats uncovered few differences in chronic ethanol-induced alterations in selected GABA_A or NMDA receptor subunit peptide levels. In general, findings for both groups of females were similar to previous observations. There was no reduction in GABA_A receptor α1 subunit levels in cerebral cortex in either intact or ovariectomized female rats, in contrast to the significant reduction observed in male rats. In addition, both intact and ovariectomized female rats had increased levels of the NMDA NR1 subunit in cerebral cortex and hypothalamus, but not in hippocampus, whereas ethanol dependent male rats displayed significant increases in the NR1 subunit only in hippocampus. Radioligand binding analysis with [³⁵S]TBPS found no differences in modulation of the GABA_A receptor by neuroactive steroids between ethanol dependent male, intact female or ovariectomized female rats. Seizure susceptibility was not different between intact or ovariectomized female rats during ethanol withdrawal. We did observe differential effects on brain allopregnanolone and plasma corticosterone levels between ethanol dependent intact and ovariectomized female rats, suggesting that ovarian steroids influence HPA axis adaptations to prolonged ethanol exposure. Overall, these data suggest that ovarian steroids do not significantly impact the gender selective alterations of GABA_A and NMDA receptors associated with ethanol dependence.
Removal of hydroxyl radical facilitates Ca<sup>2+</sup>-dependent [<sup>3</sup>H]GABA release by peroxynitrite
1998, Molecular Brain Research
We investigated mechanisms for enhancement of peroxynitrite (OONO⁻; 5 μM)-evoked [ $^{3} H$ ] γ-aminobutyric acid (GABA) release. Hydroxyl radical scavengers such as N,N′-dimethylthiourea (DMTU), mannitol, and uric acid, significantly increased OONO⁻-evoked [ $^{3} H$ ]GABA release, whereas urea showed no effects on the release. Removal of Ca²⁺ from incubation buffer abolished the enhancement of the release by DMTU, although DMTU showed no effects on the basal release with and without Ca²⁺ in extracellular space. These results indicate that hydroxyl radical scavengers facilitate OONO⁻-evoked [ $^{3} H$ ]GABA release dependent on Ca²⁺.
Enhancement of peroxynitrite-evoked acetylcholine release by hydroxyl radical scavengers from mouse cerebral cortical neurons
1998, Life Sciences
We investigated the effects of hydroxyl radical scavengers on peroxynitrite (OONO⁻)-evoked acetylcholine (ACh) release from mouse cerebral cortical neurons. N,N′-dimethylthiourea, a hydroxyl radical scavenger, dose-dependently increased OONO-evoked ACh release. Other hydroxyl radical scavengers such as uric acid and mannitol, also enhanced OONO-evoked ACh release, although these enhancing effects were not found in the absence of OONO⁻. In addition, OONO⁻-induced [⁴⁵Ca²⁺]influx was significantly facilitated by the scavengers, whereas no effects of the scavengers on [⁴⁵Ca²⁺]influx was observed in the absence of OONO⁻. These results indicate that hydroxyl radical scavengers enhance OONO⁻ -evoked ACh release via the facilitation of OONO -induced [⁴⁵Ca²⁺]influx.

View all citing articles on Scopus

View full text

Research reportGABAA receptor stimulation enhances NMDA-induced Ca2+ influx in mouse cerebral cortical neurons in primary culture

Abstract

Int. J. Dev. Neurosci.

Trends Neurosci.

Neuron

Dev. Brain Res.

Brain Res.

Dev. Brain Res.

Brain Res.

J. Biol. Chem.

Trends Pharmacol. Sci.

Dev. Brain Res.

Jpn. J. Pharmacol.

Int. J. Dev. Neurosci.

Trends Neurosci.

Trends Pharmacol. Sci.

Neuron

Research report
GABA_A receptor stimulation enhances NMDA-induced Ca²⁺ influx in mouse cerebral cortical neurons in primary culture