Spontaneous Synaptic Activity Research Articles

Na+,K+-ATPase Functionally Interacts with the Plasma Membrane Na+,Ca2+ Exchanger to Prevent Ca2+ Overload and Neuronal Apoptosis in Excitotoxic Stress

Using a fluorescent viability assay, immunocytochemistry, patch-clamp recordings, and Ca(2+) imaging analysis, we report that ouabain, a specific ligand of the Na(+),K(+)-ATPase cardiac glycoside binding site, can prevent glutamate receptor agonist-induced apoptosis in cultured rat cortical neurons. In our model of excitotoxicity, a 240-min exposure to 30 μM N-methyl-d-aspartate (NMDA) or kainate caused apoptosis in ∼50% of neurons. These effects were accompanied by a significant decrease in the number of neurons that were immunopositive for the antiapoptotic peptide Bcl-2. Apoptotic injury was completely prevented when the agonists were applied together with 0.1 or 1 nM ouabain, resulting in a greater survival of neurons, and the percentage of neurons expressing Bcl-2 remained similar to those obtained without agonist treatments. In addition, subnanomolar concentrations of ouabain prevented the increase of spontaneous excitatory postsynaptic current's frequency and the intracellular Ca(2+) overload induced by excitotoxic insults. Loading neurons with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or inhibition of the plasma membrane Na(+),Ca(2+)-exchanger by 2-(2-(4-(4-nitrobenzyloxy)phenyl)ethyl)isothiourea methanesulfonate (KB-R7943) eliminated ouabain's effects on NMDA- or kainite-evoked enhancement of spontaneous synaptic activity. Our data suggest that during excitotoxic insults ouabain accelerates Ca(2+) extrusion from neurons via the Na(+),Ca(2+) exchanger. Because intracellular Ca(2+) accumulation caused by the activation of glutamate receptors and boosted synaptic activity represents a key factor in triggering neuronal apoptosis, up-regulation of Ca(2+) extrusion abolishes its development. These antiapoptotic effects are independent of Na(+),K(+)-ATPase ion transport function and are initiated by concentrations of ouabain that are within the range of an endogenous analog, suggesting a novel functional role for Na(+),K(+)-ATPase in neuroprotection.

L-aspartate effects on single neurons and interactions with glutamate in striatal slice preparation from chicken brain

There is an accumulating evidence for a transmitter role of l-aspartate (l-Asp) in various brain regions. Recent studies from our laboratory have indicated that l-Asp is present in excitatory synapses of the striatum/nucl. accumbens of domestic chicks where it is co-released with l-glutamate (l-Glu) from axon terminals. Here we provide data on the postsynaptic effects of l-Asp alongside with l-Glu in striatal slices from chicken (1- to 10-day-old) using visually guided patch-clamp technique.l-Asp and l-Glu produced similar dose-dependent inward currents and an increase in spontaneous synaptic activity in all of the recorded striatal neurons. In the presence of TTX both the NMDA receptor antagonist D-AP5 and the AMPA/kainate receptor antagonist CNQX reduced and the co-application of these two antagonists almost abolished the postsynaptic effects of l-Asp and l-Glu in a reversible manner. Testing the interactions of l-Asp and l-Glu in these striatal neurons we found that co-application of l-Asp and l-Glut produced significantly larger inward currents than l-Asp or l-Glut alone.Our data are the first to demonstrate that l-Asp can induce postsynaptic effects on the chicken striatal neurons. These effects are mediated by both NMDA and non-NMDA type ionotropic glutamate receptors and are similar to those evoked by l-Glu. In addition our results show that co-application of l-Asp and l-Glut facilitates each other's effect, which is at least in part an excitatory amino acid (EAA) transporter dependent process. This phenomenon may explain the biological importance of the two EAAs with apparently similar postsynaptic activities in the same brain region.

Maternal Loss of Ube3a Produces an Excitatory/Inhibitory Imbalance through Neuron Type-Specific Synaptic Defects

Angelman syndrome (AS) is a neurodevelopmental disorder caused by loss of the maternally inherited allele of UBE3A. AS model mice, which carry a maternal Ube3a null mutation (Ube3a(m-/p+)), recapitulate major features of AS in humans, including enhanced seizure susceptibility. Excitatory neurotransmission onto neocortical pyramidal neurons is diminished in Ube3a(m-/p+) mice, seemingly at odds with enhanced seizure susceptibility. We show here that inhibitory drive onto neocortical pyramidal neurons is more severely decreased in Ube3a(m-/p+) mice. This inhibitory deficit follows the loss of excitatory inputs and appears to arise from defective presynaptic vesicle cycling in multiple interneuron populations. In contrast, excitatory and inhibitory synaptic inputs onto inhibitory interneurons are largely normal. Our results indicate that there are neuron type-specific synaptic deficits in Ube3a(m-/p+) mice despite the presence of Ube3a in all neurons. These deficits result in excitatory/inhibitory imbalance at cellular and circuit levels and may contribute to seizure susceptibility in AS.

TRPV1 in GABAergic Interneurons Mediates Neuropathic Mechanical Allodynia and Disinhibition of the Nociceptive Circuitry in the Spinal Cord

Neuropathic pain and allodynia may arise from sensitization of central circuits. We report a mechanism of disinhibition-based central sensitization resulting from long-term depression (LTD) of GABAergic interneurons as a consequence of TRPV1 activation in the spinal cord. Intrathecal administration of TRPV1 agonists led to mechanical allodynia that was not dependent on peripheral TRPV1 neurons. TRPV1 was functionally expressed in GABAergic spinal interneurons and activation of spinal TRPV1 resulted in LTD of excitatory inputs and a reduction of inhibitory signaling to spinothalamic tract (STT) projection neurons. Mechanical hypersensitivity after peripheral nerve injury was attenuated in TRPV1(-/-) mice but not in mice lacking TRPV1-expressing peripheral neurons. Mechanical pain was reversed by a spinally applied TRPV1 antagonist while avoiding the hyperthermic side effect of systemic treatment. Our results demonstrate that spinal TRPV1 plays a critical role as a synaptic regulator and suggest the utility of central nervous system-specific TRPV1 antagonists for treating neuropathic pain.

Short‐term field stimulation mimics synaptic maturation of hippocampal synapses

Many aspects of synaptic transmission are modified during development, reflecting not only the consequence of developmental programmes of gene expression, but also the effects of ongoing neural activity. We investigated the role of synaptic activity in the maturation of Schaffer collateral (SC)-CA1 synapses using sustained low frequency field stimulation of acute brain slices. Between postnatal days 4-6 and 14-16, mouse SC-CA1 synapses in naïve slices showed a developmental decrease in the probability of transmitter release (P(r)) and an increase in the contribution of GluN2A (NR2A) subunits to the NMDA receptor-mediated excitatory postsynaptic current (EPSC). Surprisingly, these developmental changes could be mimicked by short term (4 h) in vitro synaptic activity in slices taken from postnatal days (PND) 4-6 mice. However, different activity levels were required to alter release probability compared to the NMDA receptor subunit composition. Spontaneous synaptic activity was sufficient to alter the NMDA receptor subunit composition, but sustained low-frequency field stimulation of the brain slice (0.1 Hz, 4 h) was necessary to reduce release probability, as assessed 1 h following the cessation of stimulation. The protein synthesis inhibitor anisomycin blocked the effect of field stimulation on release probability. These results indicate that features of mature excitatory synapses can be rapidly induced in immature neurons. The activity dependence of the P(r) and NMDA receptor subunit composition serves as a sensitive indicator of prior neural activity, and provides dual mechanisms for homeostatic control of excitatory synaptic efficacy.

Plasticity of spontaneous excitatory and inhibitory synaptic activity in morphologically defined vestibular nuclei neurons during early vestibular compensation

After unilateral peripheral vestibular lesions, the brain plasticity underlying early recovery from the static symptoms is not fully understood. Principal cells of the chick tangential nucleus offer a subset of morphologically defined vestibular nuclei neurons to study functional changes after vestibular lesions. Chickens show posture and balance deficits immediately after unilateral vestibular ganglionectomy (UVG), but by 3 days most subjects begin to recover, although some remain uncompensated. With the use of whole cell voltage-clamp, spontaneous excitatory and inhibitory postsynaptic currents (sEPSCs and sIPSCs) and miniature excitatory and inhibitory postsynaptic currents (mEPSCs and mIPSCs) were recorded from principal cells in brain slices 1 and 3 days after UVG. One day after UVG, sEPSC frequency increased on the lesion side and remained elevated at 3 days in uncompensated chickens only. Also by 3 days, sIPSC frequency increased on the lesion side in all operated chickens due to major increases in GABAergic events. Significant change also occurred in decay time of the events. To determine whether fluctuations in frequency and kinetics influenced overall excitatory or inhibitory synaptic drive, synaptic charge transfer was calculated. Principal cells showed significant increase in excitatory synaptic charge transfer only on the lesion side of uncompensated chickens. Thus compensation continues when synaptic charge transfer is in balance bilaterally. Furthermore, excessive excitatory drive in principal cells on the lesion side may prevent vestibular compensation. Altogether, this work is important for it defines the time course and excitatory and inhibitory nature of changing spontaneous synaptic inputs to a morphologically defined subset of vestibular nuclei neurons during critical early stages of recovery after UVG.

Basic Concepts in Understanding Recovery of Function in Vestibular Reflex Networks during Vestibular Compensation

Unilateral peripheral vestibular lesions produce a syndrome of oculomotor and postural deficits with the symptoms at rest, the static symptoms, partially or completely normalizing shortly after the lesion due to a process known as vestibular compensation. The symptoms are thought to result from changes in the activity of vestibular sensorimotor reflexes. Since the vestibular nuclei must be intact for recovery to occur, many investigations have focused on studying these neurons after lesions. At present, the neuronal plasticity underlying early recovery from the static symptoms is not fully understood. Here we propose that knowledge of the reflex identity and input–output connections of the recorded neurons is essential to link the responses to animal behavior. We further propose that the cellular mechanisms underlying vestibular compensation can be sorted out by characterizing the synaptic responses and time course for change in morphologically defined subsets of vestibular reflex projection neurons. Accordingly, this review focuses on the perspective gained by performing electrophysiological and immunolabeling studies on a specific subset of morphologically defined, glutamatergic vestibular reflex projection neurons, the principal cells of the chick tangential nucleus. Reference is made to pertinent findings from other studies on vestibular nuclei neurons, but no comprehensive review of the literature is intended since broad reviews already exist. From recording excitatory and inhibitory spontaneous synaptic activity in principal cells, we find that the rebalancing of excitatory synaptic drive bilaterally is essential for vestibular compensation to proceed. This work is important for it defines for the first time the excitatory and inhibitory nature of the changing synaptic inputs and the time course for changes in a morphologically defined subset of vestibular reflex projection neurons during early stages of vestibular compensation.

High temperatures alter physiological properties of pyramidal cells and inhibitory interneurons in hippocampus

Temperature has multiple effects on neurons, yet little is known about the effects of high temperature on the physiology of mammalian central neurons. Hyperthermia can influence behavior and cause febrile seizures. We studied the effects of acute hyperthermia on the immature hippocampus in vitro by recording from pyramidal neurons and inhibitory oriens-lacunosum moleculare (O-LM) interneurons (identified by green fluorescent protein (GFP) expression in the GIN mouse line). Warming to 41°C caused depolarization, spontaneous action potentials, reduced input resistance and membrane time constant, and increased spontaneous synaptic activity of most pyramidal cells and O-LM interneurons. Pyramidal neurons of area CA3 were more strongly excited by hyperthermia than those of area CA1. About 90% of O-LM interneurons in both CA1 and CA3 increased their firing rates at hyperthermic temperatures; interneurons in CA3 fired faster than those in CA1 on average. Blockade of fast synaptic transmission did not abolish the effect of hyperthermia on neuronal excitability. Our results suggest that hyperthermia increases hippocampal excitability, particularly in seizure-prone area CA3, by altering the intrinsic membrane properties of pyramidal cells and interneurons.

Identification of Minimal Neuronal Networks Involved in Flexor-Extensor Alternation in the Mammalian Spinal Cord

Neural networks in the spinal cord control two basic features of locomotor movements: rhythm generation and pattern generation. Rhythm generation is generally considered to be dependent on glutamatergic excitatory neurons. Pattern generation involves neural circuits controlling left-right alternation, which has been described in great detail, and flexor-extensor alternation, which remains poorly understood. Here, we use a mouse model in which glutamatergic neurotransmission has been ablated in the locomotor region of the spinal cord. The isolated invitro spinal cord from these mice produces locomotor-like activity-when stimulated with neuroactive substances-with prominent flexor-extensor alternation. Under these conditions, unlike in control mice, networks of inhibitory interneurons generate the rhythmic activity. In the absence of glutamatergic synaptic transmission, the flexor-extensor alternation appears to be generated by Ia inhibitory interneurons, which mediate reciprocal inhibition from muscle proprioceptors to antagonist motor neurons. Our study defines a minimal inhibitory network that is needed to produce flexor-extensor alternation during locomotion.

Pre & Postsynaptic Tuning of Action Potential Timing by Spontaneous GABAergic Activity

Frequency and timing of action potential discharge are key elements for coding and transfer of information between neurons. The nature and location of the synaptic contacts, the biophysical parameters of the receptor-operated channels and their kinetics of activation are major determinants of the firing behaviour of each individual neuron. Ultimately the intrinsic excitability of each neuron determines the input-output function. Here we evaluate the influence of spontaneous GABAergic synaptic activity on the timing of action potentials in Layer 2/3 pyramidal neurones in acute brain slices from the somatosensory cortex of young rats. Somatic dynamic current injection to mimic synaptic input events was employed, together with a simple computational model that reproduce subthreshold membrane properties. Besides the well-documented control of neuronal excitability, spontaneous background GABAergic activity has a major detrimental effect on spike timing. In fact, GABAA receptors tune the relationship between the excitability and fidelity of pyramidal neurons via a postsynaptic (the reversal potential for GABAA activity) and a presynaptic (the frequency of spontaneous activity) mechanism. GABAergic activity can decrease or increase the excitability of pyramidal neurones, depending on the difference between the reversal potential for GABAA receptors and the threshold for action potential. In contrast, spike time jitter can only be increased proportionally to the difference between these two membrane potentials. Changes in excitability by background GABAergic activity can therefore only be associated with deterioration of the reliability of spike timing.

CAMP promotes differentiation of rodent neuronal progenitor cells

Numerous studies have described neuronal differentiation of neural progenitor cells derived from fetal tissue in vitro, but the biochemical mechanisms underlying this process remain largely unknown. In the present study, the role of cAMP in promoting functional maturation of neuronal progenitor cells (NPCs) from the subventricular zone (SVZ) of rodent fetal brain was investigated. NPCs were extracted from telencephalic vesicles of E14 rat embryos and then expanded in medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). A mature neuronal fate was induced by: i) withdrawal of growth factors (basal condition); ii) addition of brain-derived neurotrophic factor (BDNF); or iii) addition of isobutylmethylxantine (IBMX). Whole cell patch clamping assessed electrophysiological properties. Immunocyto - chemistry for MAP2 confirmed neuronal differentiation. Quantification of neuronal cells, and determination of their electrophysiological properties, was performed in the cited experimental groups. IBMX significantly enhanced the yield of MAP2-positive neurons in the culture system (3.7-fold increase). Application of a one-week differentiation protocol under IBMX induced functional maturity. MAP2-positive cells presented large Na- and K- voltagedependent currents, fired bursts of action potentials, and exhibited spontaneous synaptic activity. Further, IBMX proved more effective than BDNF for promoting neuronal maturation, leading to higher evoked peak currents and current densities, as well as greater yields of cells firing action potentials and presenting active synaptic contacts. Our data indicates the importance of cAMP-dependent mechanisms for maturation of neuronal progenitor cells in vitro. This knowledge can enable future manipulation of neurogenesis in vivo in order to promote the intrinsic regenerative capacity of the central nervous system.

Cannabinoids modulate spontaneous synaptic activity in retinal ganglion cells

The endocannabinoid (ECB) system has been found throughout the central nervous system and modulates cell excitability in various forms of short-term plasticity. ECBs and their receptors have also been localized to all retinal cells, and cannabinoid receptor activation has been shown to alter voltage-dependent conductances in several different retinal cell types, suggesting a possible role for cannabinoids in retinal processing. Their effects on synaptic transmission in the mammalian retina, however, have not been previously investigated. Here, we show that exogenous cannabinoids alter spontaneous synaptic transmission onto retinal ganglion cells (RGCs). Using whole-cell voltage-clamp recordings in whole-mount retinas, we measured spontaneous postsynaptic currents (SPSCs) in RGCs in adult and young (P14-P21) mice. We found that the addition of an exogenous cannabinoid agonist, WIN55212-2 (5 μM), caused a significant reversible reduction in the frequency of SPSCs. This change, however, did not alter the kinetics of the SPSCs, indicating a presynaptic locus of action. Using blockers to isolate inhibitory or excitatory currents, we found that cannabinoids significantly reduced the release probability of both GABA and glutamate, respectively. While the addition of cannabinoids reduced the frequency of both GABAergic and glutamatergic SPSCs in both young and adult mice, we found that the largest effect was on GABA-mediated currents in young mice. These results suggest that the ECB system may potentially be involved in the modulation of signal transmission in the retina. Furthermore, they suggest that it might play a role in the developmental maturation of synaptic circuits, and that exogenous cannabinoids are likely able to disrupt retinal processing and consequently alter vision.

Slow oscillating population activity in developing cortical networks: models and experimental results

During early development neuronal networks express slow oscillating synchronized activity. The activity can be driven by several, not necessarily mutually exclusive, mechanisms. Each mechanism might have distinctive consequences for the phenomenology, formation, or sustainment of the early activity pattern. Here we study the emergence of the oscillatory activity in three computational models and multisite extracellular recordings that we obtained from developing cortical networks in vitro. The modeled networks consist of leaky integrate-and-fire neurons with adaptation coupled via depressing synapses, which were driven by neurons that are intrinsically bursting, intrinsically random spiking, or driven by spontaneous synaptic activity. The activity of model networks driven by intrinsically bursting cells best matched the phenomenology of 1-wk-old cultures, in which early oscillatory activity has just begun. Intrinsically bursting neurons were present in cortical cultures, but we found them only in those cultures that were younger than 3 wk in vitro. On the other hand, synaptically dependent random spiking was highest after 3 wk in vitro. In conclusion, model networks driven by intrinsically bursting cells show a good approximation of the emergent recurrent population activity in young networks, whereas the activity of more mature networks seems to be better explained by spontaneous synaptic activity. Moreover, similar to previous experimental observations, distributed stimulation in the model was more effective in suppressing population bursts than repeated stimulation of the same neurons. This observation can be explained by an effective depression of a larger fraction of synapses by distributed stimulation.

Allostatic regulation of neuronal excitability by transient ischemia

During development and learning, a neuron actively maintains a stable state of excitability by adjusting the strength of its synaptic inputs (Turrigiano and Nelson, 2004). This homeostatic synaptic plasticity ensures that a neuron maintains a set-point spike firing rate when the overall synaptic input is increasing or decreasing (Turrigiano, 2008). Beside this homeostatic synaptic plasticity, neurons also reveal a homeostatic plasticity of neuronal intrinsic excitability by regulating voltage-dependent ion channels. When spontaneous activity is blocked for 2 days with tetrodotoxin (TTX), cortical neurons in primary cell culture increase their intrinsic excitability by a selective up-regulation of the voltage-dependent sodium current and a decrease of a voltage-dependent potassium current (Desai et al, 1999). This differential regulation of inward and outward currents increases intrinsic neuronal excitability and restores the neuron's set-point spike firing rate. A similar homeostatic stabilization of firing rates has been reported in embryonic motoneurons in vivo when spontaneous synaptic activity is reduced for 12 hours. Here, firing rates were restored by an increase in voltage-gated sodium current and a decrease in transient (IA) and calcium-dependent potassium currents (Wilhelm et al, 2009).

Staufen 2 regulates mGluR long-term depression and Map1b mRNA distribution in hippocampal neurons

The two members of the Staufen family of RNA-binding proteins, Stau1 and Stau2, are present in distinct ribonucleoprotein complexes and associate with different mRNAs. Stau1 is required for protein synthesis-dependent long-term potentiation (L-LTP) in hippocampal pyramidal cells. However, the role of Stau2 in synaptic plasticity remains unexplored. We found that unlike Stau1, Stau2 is not required for L-LTP. In contrast, Stau2, but not Stau1, is necessary for DHPG-induced protein synthesis-dependent long-term depression (mGluR-LTD). While Stau2 is involved in early development of spines, its down-regulation does not alter spine morphology or spontaneous miniature synaptic activity in older cultures where LTD occurs. In addition, Stau2, but not Stau1, knockdown reduces the dendritic localization of Map1b mRNA, a specific transcript involved in mGluR-LTD. Moreover, mGluR stimulation with DHPG induces Map1b, but not Map2, mRNA dissociation from mRNA granules containing Stau2 and the ribosomal protein P0. This dissociation was not observed in cells in which Stau2 was depleted. Finally, Stau2 knockdown reduces basal Map1b protein expression in dendrites and prevents DHPG-induced increases in dendritic Map1b protein level. We suggest a role for Stau2 in the generation and regulation of Map1b mRNA containing granules that are required for mGluR-LTD.

Effects of L-aspartate on single neurons in striatal slice preparation from chicken brain

Event Abstract Back to Event Effects of L-aspartate on single neurons in striatal slice preparation from chicken brain D. Balázs1, A. Csillag1 and G. Gerber1* 1 Semmelweis University, Dept. of Anatomy, Histology and Embryology, Hungary There is accumulating evidence for a transmitter role of L-aspartate (L-Asp) in various brain regions. Recent studies from our laboratory have indicated that L-Asp is present in excitatory synapses of the striatum/nucl. accumbens of domestic chicks where it is co-released with L-glutamate (L-Glu) from axon terminals. Here we provide data on the postsynaptic effects of L-Asp alongside with L-Glu in striatal slices from chicken (1- to 5-day-old) using visually guided patch-clamp technique. Bath application L-Asp and L-Glu produced similar dose-dependent inward currents and an increase in spontaneous synaptic activity with a threshold concentration around 200 μM in all of the recorded striatal neurons. The amplitude of the inward currents were not substantially modified when the slices were perfused with TTX-containing (1 μM) Krebs solution to block Na spikes and synaptic transmission suggesting a predominant postsynaptic effect of both excitatory amino acids. In the presence of TTX both the NMDA receptor antagonist D-AP5 (25-100 μM) and the AMPA/kainate receptor antagonist CNQX (10-20 μM) reduced and the co-application of these two antagonists abolished the postsynaptic effects of L-Asp and L-Glu in a reversible manner. The data are the first to demonstrate that L-Asp can induce postsynaptic effects on the chicken striatal neurons. These effects are mediated by both NMDA and non-NMDA type ionotropic glutamate receptors and are similar to those evoked by L-Glu. OTKA 73219 Keywords: Molecular and cellular neurobiology, Neuroscience Conference: 13th Conference of the Hungarian Neuroscience Society (MITT), Budapest, Hungary, 20 Jan - 22 Jan, 2011. Presentation Type: Abstract Topic: Molecular and cellular neurobiology Citation: Balázs D, Csillag A and Gerber G (2011). Effects of L-aspartate on single neurons in striatal slice preparation from chicken brain. Front. Neurosci. Conference Abstract: 13th Conference of the Hungarian Neuroscience Society (MITT). doi: 10.3389/conf.fnins.2011.84.00087 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 03 Mar 2011; Published Online: 23 Mar 2011. * Correspondence: Dr. G. Gerber, Semmelweis University, Dept. of Anatomy, Histology and Embryology, Budapest, Hungary, gerber@ana.sote.hu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers D. Balázs A. Csillag G. Gerber Google D. Balázs A. Csillag G. Gerber Google Scholar D. Balázs A. Csillag G. Gerber PubMed D. Balázs A. Csillag G. Gerber Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Light-evoked synaptic activity of retinal ganglion and amacrine cells is regulated in developing mouse retina

Recent studies have shown a continued maturation of visual responsiveness and synaptic activity of retina after eye opening, including the size of receptive fields of retinal ganglion cells (RGCs), light-evoked synaptic output of RGCs, bipolar cell spontaneous synaptic inputs to RGCs, and the synaptic connections between RGCs and ON and OFF bipolar cells. Light deprivation retarded some of these age-dependent changes. However, many other functional and morphological features of RGCs are not sensitive to visual experience. To determine whether light-evoked synaptic responses of RGCs undergo developmental change, we directly examined the light-evoked synaptic inputs from ON and OFF synaptic pathways to RGCs in developing retinas, and found that both light-evoked excitatory and inhibitory synaptic currents decreased, but not increased, with age. We also examined the light-evoked synaptic inputs from ON and OFF synaptic pathways to amacrine cells in developing retinas and found that the light-evoked synaptic input of amacrine cells is also downregulated in developing mouse retina. Different from the developmental changes of RGC spontaneous synaptic activity, dark rearing has little effect on the developmental changes of light-evoked synaptic activity of both RGCs and amacrine cells. Therefore, we concluded that the synaptic mechanisms mediating spontaneous and light-evoked synaptic activity of RGCs and amacrine cells are likely to be different.

Characterization of spontaneous network-driven synaptic activity in rat hippocampal slice cultures

A particular characteristic of the neonatal hippocampus is the presence of spontaneous network-driven oscillatory events, the so-called giant depolarizing potentials (GDPs). GDPs depend on the interplay between GABA and glutamate. Early in development, GABA, acting on GABAA receptors, depolarizes neuronal membranes via a Cl- efflux. Glutamate, via AMPA receptors, generates a positive feedback needed for neuronal synchronization. The depolarizing action of GABA ensures calcium entry through NMDA receptors and voltage-gated calcium channels. The whole cell configuration of the patch clamp technique (in current and voltage clamp) was used to assess whether GDPs were present also in hippocampal slice cultures. This would constitute a good model to be used for imaging and genomic approaches. GDP-like activity started appearing during the second week in culture and disappeared during the third postnatal week. GDPs occurred at the frequency 0.14 ± 0.03 Hz (n = 15). GDPs were synchronous in pair recordings from pyramidal neurons (CA1 and CA3, CA3 and CA3) and pyramidal neurons and interneurons. They reversed polarity at –35 ± 2 mV (n = 9) and were blocked by bicuculline (10 µM) or DNQX (20 µM). In the presence of bicuculline, interictal burst occurred at the frequency of 0.013 ± 0.003 Hz (n = 6). They reversed polarity at -6 ± 3 mV (n = 6) and they were blocked by DNQX (20 µM), suggesting that they were mediated by AMPA/kainate receptors. Bicuculline also blocked spontaneously occurring spike-like events observed in cell attach recordings. In additional experiments performed with gramicidin-perforated patch, the reversal potential of GDPs was –42 ± 3 mV (n = 3). These data suggest that GDP like activity recorded from organotypic slice cultures displays a pattern very similar to that observed in acute slices obtained from neonatal rats and suggest that, at least in the majority of neurons, the action of GABA is still depolarizing and excitatory.

Altered function of glutamatergic cortico-striatal synapses causes output pathway abnormalities in a chronic model of parkinsonism

Objective In Parkinson's disease, chronic striatal dopamine depletion results in over-activity and under-activity of the indirect and direct striatal output pathways respectively. In this study, we investigated changes in the function of glutamatergic cortico-striatal synapses that contribute to abnormalities in striatal efferents. Methods Whole-cell recordings were performed in striatal slices prepared from adult bacterial artificial chromosome mice, chronically lesioned with 6-hydroxydopamine (6-OHDA). Paired pulse facilitation, spontaneous synaptic activity, the ratio of AMPAR to NMDAR-mediated components of excitatory postsynaptic currents, AMPAR and NMDAR kinetics, current–voltage relationship and intrinsic membrane properties were assessed in indirect and direct pathway medium spiny neurons (MSNs), which were identified on the basis of expression of GFP, driven by the promoters of A2A or D1 receptor expression. The trajectory of striatal efferents, with respect to selective targeting of the globus pallidus and substantia nigra was also compared in sham-operated versus 6-OHDA-lesioned mice. Results Dopamine depletion did not affect the number of pathway specific output neurons or the trajectory of striatal outputs. In sham-operated animals, cortico-striatal synapses of both striatal efferent populations exhibited paired pulse facilitation and similar ratios of AMPAR to NMDAR-mediated components of excitatory postsynaptic currents. Following striatal dopamine depletion, indirect pathway neurons exhibited decreased levels of paired pulse facilitation, enhanced sensitivity to presynaptic stimulation and an increase in the relative contribution of NMDAR to the EPSC but no change in spontaneous synaptic activity. In sham-operated mice, neurons of the direct pathway exhibited lower firing frequency compared to the indirect pathway following current injection. However, in 6-OHDA-lesioned mice, in the direct pathway, firing threshold was reduced, spike frequency adaptation developed and the frequency of spontaneous activity was also reduced. In addition, changes in the properties of NMDAR kinetics suggest that these receptors were desensitised. Discussion Increased synchronicity between pre and postsynaptic neurons, as indicated by decreased paired pulse facilitation, and increased sensitivity to extracellular stimulation, combined with an increase in the contribution of NMDARs to the EPSC at cortico-striatal synapses, may contribute to the over-activity of indirect pathway neurons in the parkinsonian striatum. In contrast, a decrease in spontaneous activity, postsynaptic desensitisation to excitatory stimuli and spike frequency adaptation of cortico-striatal synapses may underlie under-activity of the direct pathway.

Comparative study of cellular and synaptic abnormalities in brain tissue samples from pediatric tuberous sclerosis complex and cortical dysplasia type II

Tuberous sclerosis complex (TSC) and severe cortical dysplasia (CD), or CD type II according to Palmini classification, share histopathologic similarities, specifically the presence of cytomegalic neurons and balloon cells. In this study we examined the morphologic and electrophysiologic properties of cells in cortical tissue samples from pediatric patients with TSC and CD type II who underwent surgery for pharmacoresistant epilepsy. Normal-appearing pyramidal neurons from TSC and CD type II cases had similar passive membrane properties. However, the frequency of excitatory postsynaptic currents (EPSCs) was higher in neurons from TSC compared to severe CD cases, particularly the frequency of medium- and large-amplitude synaptic events. In addition, EPSCs rise and decay times were slower in normal cells from TSC compared to severe CD cases. Balloon cells were found more frequently in TSC cases, whereas cytomegalic pyramidal neurons occurred more often in CD type II cases. Both cell types were similar morphologically and electrophysiologically in TSC and severe CD. These results suggest that even though the histopathology in TSC and severe CD is similar, there are subtle differences in spontaneous synaptic activity and topographic distribution of abnormal cells. These differences may contribute to variable mechanisms of epileptogenesis in patients with TSC compared with CD type II.