The monosynaptic connection: the modulating effect of opioid peptides on the plasticity of presynaptic neurons and identified synapses]

Study of opioid peptides (leucine-enkephalin and methionine-enkephalin) action on plastic properties of the system of monosynaptically connected neurones LPa7--LPa3, PPa3 and LPa8--LPa3, PPa3 was conducted in the snail brain. It has been shown that all three links in the system studied (presynaptic neurone, postsynaptic neurone and synapse) manifest one and the same type of plasticity--habituation to rhythmic stimulation. Enkephalins have a modulating action on plastic properties of the presynaptic neurone and synapse: they retard the habituation of the presynaptic neurone to intracellular stimulation and retard the development of habituation at synaptic level. However, changes in the character of postsynaptic response in the presence of enkephalins are not a direct consequence of their influence on plastic properties of the presynaptic neurone. Besides, enkephalines reduce the effectiveness of synaptic transmission in the given system: they reduce EPSP duration in the postsynaptic neurone.     

Effects of FMRFamide on the activity of command neurons in defensive behavior of fed and fasting snails, Helix pomatia

FMRFamide (Phe-Met-Arg-Phe-NH2) micropneumophoresis changed bimodally the activity of LPa2, LPa3, PPa2 and PPa3 neurones in fasting and fed Helix pomatia. In fasting creatures peptide application elicited hyperpolarization and decreased the neuronal membrane excitability and responses to tactile stimulation. In fed snails peptide application caused depolarization, decreased membrane resistance and increased the neuronal membrane excitability and responses to tactile stimulation. Neurophysiological mechanisms underlying FMRFamide effects on feeding and defense behaviour are discussed.     

Ionic mechanisms of the transmembrane current evoked by injection of cyclic amp into identified Helix pomatia neurons

Ionic mechanisms of the transmembrane current evoked by injection of cyclic AMP into identified neurons ofHelix pomatia were investigated by the voltage clamp method. Injection of cyclic AMP into neurons RPa3, LPa2, LPa3, and LPl1 was shown to cause the development of a two-component transmembrane (cyclic AMP) current. The current-voltage characteristic curve of the early component is linear in the region from –40 to –90 mV; the reversal potential of the early component, determined by extrapolation, lies between –5 and +20 mV; the current-voltage characteristic curve of the late component also is linear and has a reversal potential between –55 and –60 mV. A decrease in the sodium concentration in the external medium from 100 to 25 mM led to a decrease in amplitude of the cyclic AMP current and to a shift of the reversal potential for the early component by 30–32 mV toward hyperpolarization. It is suggested that the early component of the cyclic AMP current in neurons RPa3, LPa2, LPa3, and LPl1 is associated with an increase in permeability of the neuron membrane chiefly for sodium ions, whereas the late component is correspondingly connected with permeability for potassium ions. Injection of cyclic AMP also caused the appearance of a transmembrane inward current in neuron LPa8, but it was independent of the holding potential and was unaccompanied by any change in membrane permeability. It is suggested that this current may be due to a change in the activity of the electrogenic ion pump.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 526–532, September–October, 1980.     

Rf 0.58 protein in Helix command neurons during learning

At early stages of aversive conditioning in Helix, a most considerable increase in the acid brain-specific protein Rf 0.58 occurred in LPa3 and PPa3 neurones. Later the content of this protein decreased in the PPa3 but went on increasing in LPa3. In sham learning, the protein content did not increase so obviously. Hence the protein Rf 0.58 metabolism in individual neurones of the snail CNS correlates with the draw step of receptor and effector fields in avoidance conditioning.     

Postsynaptic potentials in the myocardium of snails in the genus Helix

Cardioregulating neurones in the right parietal and visceral ganglia of the snail evoke postsynaptic potentials of various duration, amplitude and polarity in the auricular and ventricular myocardium. Inhibitory neurones with a marked background activity (1-2 imp/s) evoke IPSPs with a duration of 150-200 msec and a latent period of 160-220 msec in the auricle, these potentials being blocked by tubocurarine. EPSPs of approximately the same duration may be recorded in the ventricle during stimulation of the commanding neurones of the pneumostome LPa3 and PPa/3, as well as unidentified neurones. Action potentials in some other identified cardiostimulating neurones (PPa7, V1, V6) induce slow and sustained depolarization in the myocardium. Functional specificity of elements within fast and slow regulatory systems is suggested: discrete IPSPs and EPSPs account mainly for coordination of the systolic contractions of the auricle and ventricle, whereas long-lasting PSPs affect the frequency and intensity of the whole heart.     

Contribution of a humoral factor to postsynaptic sensitization in heterosynaptic facilitation

It was shown that heterosynaptic facilitation develops in the cerebral ganglia giant neurons of the freshwater gastropod molluskPlanorbis corn eus due to diffuse neurohumoral influences on pre- and postsynaptic structures and not local synaptic action on presynaptic mechanisms. It was also found that n-cholinergic synaptic mechanisms come under this facilitatory influence. Serotonin is the source of facilitation in neurons of bothPlanorbis corneus cerebral ganglion and those of the aplysia abdominal ganglion. Seeing that: a) conditioning stimuli facilitate the effects produced by iontophoretic acetylcholine application, as well as n-cholinergic synaptic transmission and b) the amplitude of EPSP and acetylcholine potential increase 4–6 times during facilitation when the input impedance of the post-synaptic membrane is increased by just 20%, it was deduced that the postsynaptic membrane of the giant neuron makes a significant contribution to heterosynaptic facilitation of the sensitization of n-cholinergic receptors. The part played by n-cholinergic receptors of the postsynaptic membrane in heterosynaptic facilitation and conditioned reflex habituation is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 18, No. 2, pp. 250–259, March–April, 1986.     

The synaptic scaffold protein MPP2 interacts with GABAA receptors at the periphery of the postsynaptic density of glutamatergic synapses

Recent advances in imaging technology have highlighted that scaffold proteins and receptors are arranged in subsynaptic nanodomains. The synaptic membrane-associated guanylate kinase (MAGUK) scaffold protein membrane protein palmitoylated 2 (MPP2) is a component of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor–associated protein complexes and also binds to the synaptic cell adhesion molecule SynCAM 1. Using superresolution imaging, we show that—like SynCAM 1—MPP2 is situated at the periphery of the postsynaptic density (PSD). In order to explore MPP2-associated protein complexes, we used a quantitative comparative proteomics approach and identified multiple γ-aminobutyric acid (GABA)_A receptor subunits among novel synaptic MPP2 interactors. In line with a scaffold function for MPP2 in the assembly and/or modulation of intact GABA_A receptors, manipulating MPP2 expression had effects on inhibitory synaptic transmission. We further show that GABA_A receptors are found together with MPP2 in a subset of dendritic spines and thus highlight MPP2 as a scaffold that serves as an adaptor molecule, linking peripheral synaptic elements critical for inhibitory regulation to central structures at the PSD of glutamatergic synapses.

This study shows that the MAGUK scaffold protein MPP2 is located at the periphery of postsynaptic densities in excitatory neurons, where it interacts with GABA-A receptors, thereby serving as a functional adaptor that links excitatory and inhibitory components of synaptic transmission at glutamatergic synapses.     

GABAB receptor modulation of synaptic function

Neuromodulators have complex effects on both the presynaptic release and postsynaptic detection of neurotransmitters. Here we describe recent advances in our understanding of synaptic modulation by metabotropic GABAB receptors. By inhibiting multivesicular release from the presynaptic terminal, these receptors decrease the synaptic glutamate signal. GABAB receptors also inhibit the Ca2+ permeability of NMDA receptors to decrease Ca2+ signals in postsynaptic spines. These new findings highlight the importance of GABAB receptors in regulating many aspects of synaptic transmission. They also point to novel questions about the spatiotemporal dynamics and sources of synaptic modulation in the brain.     

Giant polyfunctional neuron of the edible snail

The structure of the receptive field of LPa3 neurone and its connection with the periphery were studied on semi-intact preparations of the snail Helix pomatia. It was found that: 1) The mechano-sensitive receptive field (excitatory) of the LPa3 neurone occupies nearly the whole surface of the snail's skin and internal organs. But latencies of LPa3 reactions to mechanical stimulation, the whole field may be divided into four zones: 50 to 60 ms, 100 ms, 130 to 140 ms and 200 to 250 ms. 2) Blockade of synaptic transmission with magnesium or cobalt has shown that relaying of the signal from the mantle receptors to the LPa3 neurone takes place in the peripheral nervous system. 3) Cobalt ionophoresis showed that the LPa3 neurone gives off processes to the right and left pallial nerves, the anal nerve and, occasionally, to the cutaneous nerve. During simultaneous intracellular recording from the LPa3 neurone and extracellular from the above nerves, action potentials are in every case recorded at first in the neurone body, and then, with a 20 to 30 ms delay, in the nerves. This means that the LPa3 neurone processes joining the nerves are axons. It is suggested that the sensory inputs and wide branching structures output of the LPa3 neurone axons make it an integrating polyfunctional system.     

Regions of putative acetylcholine receptors at synaptic contacts between neurons maintained in culture and subsequently fixed in solutions containing tannic acid

Summary Spinal cord neurons from 9-day chick embryos were maintained in culture for up to 35 days and then fixed in 4% cacodylate-buffered glutaraldehyde containing 2% tannic acid. After about 15 days in culture a small percentage of the synaptic specializations present were characterized by striking electron-dense striations averaging 15 nm in width, oriented perpendicular to the postsynaptic membrane. These structures increased in frequency with time in culture (to a maximum of about 10% of all synapses in the oldest cultures); they were asymmetrical, protruding approximately 8 nm into the synaptic cleft, and more deeply (approximately 15–18 nm), into the postsynaptic cytoplasm. On the basis of earlier work by Sealock (1980) they are interpreted as concentrations of acetylcholine receptors.Similar membrane differentiations were also seen associated with active-zone areas of a few presynaptic membranes, and the possibility that these represent presynaptic acetylcholine receptors is discussed. Additional observations reported are (1) the presence of striations resembling those seen at the postsynaptic membrane in the membranes of some postsynaptic vesicles, and (2) filamentous links between the striations and cytoskeletal elements of the postsynaptic cell.     

Neural Organization of the Median Ocellus of the Dragonfly : I. Intracellular electrical activity

Intracellular responses from receptors and postsynaptic units have been recorded in the median ocellus of the dragonfly. The receptors respond to light with a graded, depolarizing potential and a single, tetrodotoxin-sensitive impulse at "on." The postsynaptic units (ocellar nerve dendrites) hyperpolarize during illumination and show a transient, depolarizing response at "off." The light-evoked slow potential responses of the postsynaptic units are not altered by the application of tetrodotoxin to the ocellus. It appears, therefore, that the graded receptor potential, which survives the application of tetrodotoxin, is responsible for mediating synaptic transmission in the ocellus. Comparison of pre- and postsynaptic slow potential activity shows (a) longer latencies in postsynaptic units by 5–20 msec, (b) enhanced photosensitivity in postsynaptic units by 1–2 log units, and (c) more transient responses in postsynaptic units. It is suggested that enhanced photosensitivity of postsynaptic activity is a result of summation of many receptors onto the postsynaptic elements, and that transients in the postsynaptic responses are related to the complex synaptic arrangements in the ocellar plexus to be described in the following paper.     

Effects of ethimizol on frog neuromuscular junction

The effects were studied of ethimizol, a substance activating memory processes, on features of synaptic transmission during experiments on frog cutaneous pectoris muscle. It was found that the presynaptic action of ethimizol consists of raising the frequency of miniature potentials, when used at a concentration of 0.5–10 mM, and modulating quantal content of synaptic transmission due to changes in binomial quantal release parameters p and n when 0.5–2 mM ethimizol was used. This substance facilitated transmission at synapses with a low initial level of transmitter release. This substance facilitated transmission at synapses with a low initial level of transmitter release. Ethimizol was also found to have a postsynaptic action, consisting of reducing amplitude at a concentration of 5–10 mM and prolonging synaptic currents and potentials when concentrations of 0.5–10 mM were used. The latter effect produced a considerable increase in the time integral of endplate potentials. The postsynaptic action of ethimizol is perhaps seen in its effects on features of postsynaptic ionic channels. The effects of ethimizol are discussed with a view to how it may act within the central nervous system as a nonspecific modulator.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 17, No. 6, pp. 757–763, November–December, 1985.     

Identification of cholinoreceptors on the soma of snail neurons RPa3 and LPa3

Identification of cholinoreceptors (CR) of the soma of neurons RPa3 and LPa3 of the snail is performed using selective cholinomimetics and cholinolytics during the recording of transmembrane ionic currents. Agonists of the nicotinic (NCR) and muscarinic (MCR) types of cholinolytics evoked a brief activation of the receptors, with the exception of carbamylcholine, followed by an irreversible blocking. All selective cholinomimetics bonded with the same membrane centers which acetylcholine (AC) activated. The nicotinic and muscarinic cholinolytics decreased the amplitude of the input current elicited by AC; however, the use of scopolamine and platyphylline was without effect. It is speculated that the soma of neurons RPa3 and LPa3 exhibits NCR and MCR which have a number of pharmacological features distinguishing them from the corresponding CR of vertebrates. The MCR of these neurons must be classed as a special subtype differing from the well-known M₁ and M₂ subtypes.M. V. Lomonosov State University of Moscow. Translated from Neirofiziologiya, Vol. 24, No. 1, pp. 77–86, January–February, 1992.     

Synaptic effects of nicotinic and muscarinic agonists in sympathetic ganglia of the frog

Superfusion of the isolated sympathetic ganglia of the frog with nicotinic agonists (suberyldicholine, tetramethylamonium, and dimethylphenylpiperazinium), as well as acetylcholine in the presence of atropine led to a brief depolarization of the neurons and blockade of synaptic transmission. The muscarinic agonists methylfurmethide (MFM) and methyldilvasen, cis^–, L(+), as well as acetylcholine elicited a stable depolarization which is not accompanied by disturbance in transmission. Oxotremorine at a concentration of 1·10^–5 M did not lead to the depolarization of the post-synaptic membrane, but at a concentration of 1·10^–6 M decreased the quantal EPSP content twofold, which indicates that the presynaptic receptors belong to the M₂ subtype. Inhibition of acetylcholinesterase significantly intensified the postsynaptic effect of MFM: a shift of the concentration-effect curve took place toward the side of lower MFM concentrations. It was shown that the post-synaptic muscarinic receptors of the ganglionic neurons possess varied sensitivity to the enantiomers of methyldilvasen and, consequently, are stereospecific. The identified functional properties of the cholinoreceptors of the ganglionic neurons explain the set of changes in synaptic transmission under conditions of the prolonged presence of a mediator in the synaptic cleft.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 20, No. 2, pp. 227–234, March–April, 1988.     

Action of ammonium acetate on central primary afferent depolarization

Experiments on anesthetized spinal cats showed that ammonium acetate, injected intravenously (2–4 mmoles/kg) inhibits the depolarization of the central endings of primary afferent fibers activated by stimulation of afferent nerves. Inhibition of primary afferent depolarization is transient in character and develops parallel with depression of postsynaptic inhibition of monosynaptic reflexes. The depression produced by the action of ammonium was not due to blocking of negative postsynaptic potentials of the dorsal surface of the spinal cord or blocking of reflex electrical discharges in the ventral spinal roots. It is suggested that depression of primary afferent depolarization is due to a decrease in the emf for synaptic ion currents producing depolarization.Allergologic Research Laboratory, Academy of Medical Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 9, No. 1, pp. 52–60, January–February, 1977.     

Homeostatic Competition between Phasic and Tonic Inhibition

The GABA_A receptors are the major inhibitory receptors in the brain and are localized at both synaptic and extrasynaptic membranes. Synaptic GABA_A receptors mediate phasic inhibition, whereas extrasynaptic GABA_A receptors mediate tonic inhibition. Both phasic and tonic inhibitions regulate neuronal activity, but whether they regulate each other is not very clear. Here, we investigated the functional interaction between synaptic and extrasynaptic GABA_A receptors through various molecular manipulations. Overexpression of extrasynaptic α6β3δ-GABA_A receptors in mouse hippocampal pyramidal neurons significantly increased tonic currents. Surprisingly, the increase of tonic inhibition was accompanied by a dramatic reduction of the phasic inhibition, suggesting a possible homeostatic regulation of the total inhibition. Overexpressing the α6 subunit alone induced an up-regulation of δ subunit expression and suppressed phasic inhibition similar to overexpressing the α6β3δ subunits. Interestingly, blocking all GABA_A receptors after overexpressing α6β3δ receptors could not restore the synaptic GABAergic transmission, suggesting that receptor activation is not required for the homeostatic interplay. Furthermore, insertion of a gephyrin-binding-site (GBS) into the α6 and δ subunits recruited α6_GBSβ3δ_GBS receptors to postsynaptic sites but failed to rescue synaptic GABAergic transmission. Thus, it is not the positional effect of extrasynaptic α6β3δ receptors that causes the down-regulation of phasic inhibition. Overexpressing α5β3γ2 subunits similarly reduced synaptic GABAergic transmission. We propose a working model that both synaptic and extrasynaptic GABA_A receptors may compete for limited receptor slots on the plasma membrane to maintain a homeostatic range of the total inhibition.     

Long-term depression-inducing stimuli promote cleavage of the synaptic adhesion molecule NGL-3 through NMDA receptors,matrix metalloproteinases and presenilin/γ-secretase

Long-term depression (LTD) reduces the functional strength of excitatory synapses through mechanisms that include the removal of AMPA glutamate receptors from the postsynaptic membrane. LTD induction is also known to result in structural changes at excitatory synapses, including the shrinkage of dendritic spines. Synaptic adhesion molecules are thought to contribute to the development, function and plasticity of neuronal synapses largely through their trans-synaptic adhesions. However, little is known about how synaptic adhesion molecules are altered during LTD. We report here that NGL-3 (netrin-G ligand-3), a postsynaptic adhesion molecule that trans-synaptically interacts with the LAR family of receptor tyrosine phosphatases and intracellularly with the postsynaptic scaffolding protein PSD-95, undergoes a proteolytic cleavage process. NGL-3 cleavage is induced by NMDA treatment in cultured neurons and low-frequency stimulation in brain slices and requires the activities of NMDA glutamate receptors, matrix metalloproteinases (MMPs) and presenilin/γ-secretase. These results suggest that NGL-3 is a novel substrate of MMPs and γ-secretase and that NGL-3 cleavage may regulate synaptic adhesion during LTD.     

Cholinergic modulation of the synaptic transmission in the frog spinal cord

It was found during experiments on isolated frog spinal cord involving extracellular recording from the dorsal roots (sucrose bridging) and intracellular recording from motoneurons by microelectrodes that 10 mM of the M-cholinomimetic arecoline produces motoneuronal depolarization which is matched by depolarizing electronic ventral root potentials and a rise in motoneuronal input resistance. Arecoline changes synaptic transmission by increasing the amplitude of postsynaptic potentials during intracellular recording and that of motoneuronal reflex discharges in the ventral roots but reduces the duration of dorsal root potentials. In the presence of arecoline, L-glutamate-induced motoneuronal response increases. Facilitation of synaptic transmission produced by arecoline in the spinal cord is bound up with cholinergic M₂- activation, since it is suppressed by atropine but not by low concentrations of pirenzipine; it is also coupled with a reduction in adenylcyclase activity. When motoneuronal postsynaptic response has been suppressed, as in the case of surplus calcium or theophylline, arecoline produces an inhibitory effect on the amplitude of motoneuronal monosynaptic reflex discharges which is suppressed by pirenzipine at a concentration of 1×10^–7 M. This would indicate the presence at the primary afferent terminals of presynaptic cholinergic M₁ receptors which mediate its inhibition of impulses of transmitter release. This effect is independent of changes in cyclic nucleotide concentration.A. M. Gorkii Medical Institute, Donetsk. Translated from Neirofiziologiya, Vol. 19, No. 3, pp. 399–405, May–June, 1987.     

mGluRs Modulate Strength and Timing of Excitatory Transmission in Hippocampal Area CA3

Excitatory transmission within hippocampal area CA3 stems from three major glutamatergic pathways: the perforant path formed by axons of layer II stellate cells in the entorhinal cortex, the mossy fiber axons originating from the dentate gyrus granule cells, and the recurrent axon collaterals of CA3 pyramidal cells. The synaptic communication of each of these pathways is modulated by metabotropic glutamate receptors that fine-tune the signal by affecting both the timing and strength of the connection. Within area CA3 of the hippocampus, group I mGluRs (mGluR1 and mGluR5) are expressed postsynaptically, whereas group II (mGluR2 and mGluR3) and III mGluRs (mGluR4, mGluR7, and mGluR8) are expressed presynaptically. Receptors from each group have been demonstrated to be required for different forms of pre- and postsynaptic long-term plasticity and also have been implicated in regulating short-term plasticity. A recent observation has demonstrated that a presynaptically expressed mGluR can affect the timing of action potentials elicited in the postsynaptic target. Interestingly, mGluRs can be distributed in a target-specific manner, such that synaptic input from one presynaptic neuron can be modulated by different receptors at each of its postsynaptic targets. Consequently, mGluRs provide a mechanism for synaptic specialization of glutamatergic transmission in the hippocampus. This review will highlight the variability in mGluR modulation of excitatory transmission within area CA3 with an emphasis on how these receptors contribute to the strength and timing of network activity within pyramidal cells and interneurons.     

Membrane-Derived Phospholipids Control Synaptic Neurotransmission and Plasticity

Synaptic communication is a dynamic process that is key to the regulation of neuronal excitability and information processing in the brain. To date, however, the molecular signals controlling synaptic dynamics have been poorly understood. Membrane-derived bioactive phospholipids are potential candidates to control short-term tuning of synaptic signaling, a plastic event essential for information processing at both the cellular and neuronal network levels in the brain. Here, we showed that phospholipids affect excitatory and inhibitory neurotransmission by different degrees, loci, and mechanisms of action. Signaling triggered by lysophosphatidic acid (LPA) evoked rapid and reversible depression of excitatory and inhibitory postsynaptic currents. At excitatory synapses, LPA-induced depression depended on LPA₁/G_αi/o-protein/phospholipase C/myosin light chain kinase cascade at the presynaptic site. LPA increased myosin light chain phosphorylation, which is known to trigger actomyosin contraction, and reduced the number of synaptic vesicles docked to active zones in excitatory boutons. At inhibitory synapses, postsynaptic LPA signaling led to dephosphorylation, and internalization of the GABA_Aγ2 subunit through the LPA₁/G_α12/13-protein/RhoA/Rho kinase/calcineurin pathway. However, LPA-induced depression of GABAergic transmission was correlated with an endocytosis-independent reduction of GABA_A receptors, possibly by GABA_Aγ2 dephosphorylation and subsequent increased lateral diffusion. Furthermore, endogenous LPA signaling, mainly via LPA₁, mediated activity-dependent inhibitory depression in a model of experimental synaptic plasticity. Finally, LPA signaling, most likely restraining the excitatory drive incoming to motoneurons, regulated performance of motor output commands, a basic brain processing task. We propose that lysophospholipids serve as potential local messengers that tune synaptic strength to precedent activity of the neuron.