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Exercise differentially regulates synaptic proteins associated to the function of BDNF.

We explored the capacity of exercise to impact select events comprising synaptic transmission under the direction of brain-derived neurotrophic factor (BDNF), which may be central to the events by which exercise potentiates synaptic function. We used a specific immunoadhesin chimera (TrkB-IgG) that mimics the BDNF receptor, TrkB, to selectively block BDNF in the hippocampus during 3 days of voluntary wheel running. We measured resultant synapsin I, synaptophysin, and syntaxin levels involved in vesicular pool formation, endocytosis, and exocytosis, respectively. Synapsin I is involved in vesicle pool formation and neurotransmitter release, synaptophysin, in the biogenesis of synaptic vesicles and budding, and syntaxin, in vesicle docking and fusion. Exercise preferentially increased synapsin I and synaptophysin levels, without affecting syntaxin. There was a positive correlation between synapsin I and synaptophysin in exercising rats and synapsin I with the amount of exercise. Blocking BDNF abrogated the exercise-induced increases in synapsin I and synatophysin, revealing that exercise regulates select properties of synaptic transmission under the direction of BDNF.

Animals↗

Significance of extracellular potassium in central respiratory control studied in the isolated brainstem-spinal cord preparation of the neonatal rat.

The significance of extracellular potassium in central respiratory control was investigated using the isolated brainstem-spinal cord preparation of the neonatal rat. Depth profiles of extracellular potassium activity ([K+])ECF in the medulla were measured with ion-sensitive microelectrodes. Although [K+]ECF increased with depth in medullary tissue during control (4 mM) and low (1 mM) potassium concentration ([K+])CSF superfusion, this gradient disappeared with higher [K+]CSF. With low [K+]CSF (1 mM), respiratory CO2 responsiveness was abolished, and increased with high [K+]CSF (8 mM). Respiratory frequency (fR) was diminished at low [K+]CSF (1 mM), and increased with elevated [K+]CSF (8 and 16 mM); with yet higher [K+]CSF (32 mM) apnea occurred after a transient increase in fR. Perforated patch recording revealed that high [K+]ECF decreased membrane resistance, depolarized membrane potential, and increased firing frequency in most of the recorded medullary neurons. High [K+]ECF also increased excitatory and inhibitory post-synaptic potentials of medullary neurons and augmented the functional connectivity among neurons. It is concluded that [K+]ECF is of importance in the maintenance of respiratory rhythm and central chemosensitivity.

Analysis of Variance↗

Co-transmitter function of ATP in central catecholaminergic neurons of the rat.

Intracellular recordings were made in a mid-pontine slice preparation of the rat brain containing the nucleus locus coeruleus. Focal electrical stimulation evoked biphasic synaptic potentials consisting of early depolarizing (d.p.s.p.) and late hyperpolarizing (i.p.s.p.) components. The alpha(2)-adrenoceptor antagonist idazoxan inhibited the i.p.s.p. without altering the d.p.s.p. All of the following experiments were carried out in the presence of kynurenic acid and picrotoxin to block the glutamatergic and GABAergic fractions of the d.p.s.p., respectively. Guanethidine, which is known to inhibit noradrenaline and ATP release from nerve terminals of postganglionic sympathetic nerves, depressed both the d.p.s.p. and the i.p.s.p. in a concentration-dependent manner. Damage of catecholaminergic nerve terminals by 6-hydroxydopamine also decreased both the d.p.s.p. and the i.p.s.p. The P2 receptor antagonist pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) depressed the d.p.s.p., whereas the i.p.s.p. remained unaffected. The further application of PPADS did not increase the depression of the d.p.s.p. by guanethidine. Superfusion with the mixed alpha-adrenoceptor agonist noradrenaline or the selective P2 receptor agonist adenosine 5'-O-(2-thiodiphosphate) inhibited both the d.p.s.p. and the i.p.s.p. The inhibitory effects of these agonists were prevented by the respective antagonists idazoxan or suramin. In the presence of suramin noradrenaline failed to inhibit the residual d.p.s.p. Superfused noradrenaline potentiated rather than inhibited responses to pressure-applied alpha,beta-methylene-ATP; superfused adenosine 5'-O-(2-thiodiphosphate) did not interact with pressure-applied noradrenaline. In conclusion, we present electrophysiological evidence for the co-release of ATP and catecholamines in the CNS. At the cell somata of neurons in the locus coeruleus, noradrenaline and ATP activate inhibitory alpha(2)-adrenoceptors and excitatory P2 receptors, respectively. In addition, inhibitory presynaptic autoreceptors of the alpha(2) and P2 types appear to regulate release of the two co-transmitters.

Adenosine Diphosphate↗

Excitatory effect of thiamin on CA1 pyramidal neurones in rat hippocampal slices in vitro.

Clinically, acute thiamin deficiency may lead to Wernicke encephalopathy and fulminant cardial beriberi. Both diseases respond to high parenteral doses of thiamin. The cofactor role of intracellular thiamin diphosphate has been thoroughly investigated, but an additional acute effect of unphosphorylated extracellular thiamin has been postulated but not elucidated. In order to investigate the role of thiamin at the membrane level in the central nervous system, a study using a well-established in vitro rat hippocampal slice model was designed. Hippocampal slices were perfused with 0.5, 0.75 and 1 mM thiamin solutions for 30 min and the pre-synaptic volley, field excitatory post-synaptic potential and population spike amplitudes were recorded continuously. The results showed an acute, excitatory effect of high-dose thiamin on hippocampal neurones by significantly increasing the number of repetitive afterdischarges. Additional experiments with low concentrations of the potassium channel blocker 4-aminopyridine showed similar findings. The results support previous evidence of thiamin affecting membrane ion channel activity, probably involving potassium channels, although the precise mechanisms of action are still unclear.

4-Aminopyridine↗

Effect of barbiturates on 'quantal' synaptic transmission in spinal motoneurones.

1. Monosynaptic excitatory post-synaptic potentials (EPSPs) produced in triceps surae motoneurones of the cat by stimulation of single afferent fibres in the muscle nerve were recorded with intracellular electrodes before and after the administration of thiopentone or pentobarbitone.2. The average amplitude of the ;unit' EPSP was 0.11-0.21 mV and remained unchanged after the administration of the barbiturates (10 mg/kg, I.V.).3. Mean quantum content (m) ranged from 1.9 to more than 5 before drug administration. The m was reduced by thiopentone (10 mg/kg, I.V.) and pentobarbitone (10 mg/kg, I.V.) by 23.1 and 24.7% respectively.4. The barbiturates, in the doses employed, produced no alterations in the input resistance of the motoneurone membrane or in the strength-duration relation obtained by passing depolarizing current pulses through the micro-electrode.5. It is concluded that the action of thiopentone and pentobarbitone, in the doses used, is confined to the presynaptic nerve terminals and results in a reduction in the amount of transmitter released by afferent impulses.

Action Potentials↗

Specific effects of alpha-D,L-aminoadipic acid on synaptic transmission in frog spinal cord.

The sucrose gap technique was employed to investigate both synaptic and amino acid evoked responses from motoneurones or primary afferents of frog spinal cord. alpha-D,L-Aminoadipic acid (alpha-D,L-AAD) selectively antagonized responses to acidic amino acids, especially aspartate. The drug was most effective in antagonizing the polysynaptic components of synaptic potentials evoked by dorsal root or lateral column stimulation but had little effect on their monosynaptic components. The ventral root dorsal root potential which is thought to be mediated by a pathway that does not involve acidic amino acids was insensitive to alpha-D,L-AAD. These data, which were confirmed by intracellular recording from motoneurones, provided further evidence for the role of acidic amino acids in polysynaptic pathways in frog spinal cord.

2-Aminoadipic Acid↗

Hydraulic analogs as teaching tools for bioelectric potentials.

Hydraulic analogs of bioelectrical potentials are proposed as teaching tools in helping students with no formal background in physics and mathematics. Membrane capacitance is simulated by a water reservoir, V, whose variable level is the membrane potential. Resting membrane potential is simulated by a large capacitance reservoir of fixed level, connected to reservoir V through a tube having the role of the electrical conductance of the membrane. Injection of electrical current into the cell is simulated by injection of water into the membrane capacitance reservoir. Reversal potentials of the end-plate potential are simulated by another water reservoir, with fixed level (analogous to the reversal potential of the end-plate potential) connected to the membrane capacitance reservoir through a tube corresponding to the acetylcholine-activated ion channels. Different phases of the synaptic potential are then described using the hydraulic analogs. Hydraulic analogs have proved, in our experience, to be an efficient tool in complementing the already established electrical equivalents.

Education, Medical↗

Synaptic organization in teleost spinal motoneurons.

Glass microelectrodes were inserted into motoneurons innervating pectoral fin muscles to record action and synaptic potentials, evoked by electrical stimulation of ventral and dorsal roots, and the medulla oblongata. Ventral root stimulation evoked a small depolarizing response which had properties compatible with those of the EPSP; its amplitude changes were graded, being increased by membrane hyperpolarization and decreased by high frequency repetitive stimulation. The latency of the response was sufficiently longer than that of the antidromic spike to allow for a monosynaptic delay. Stimulation of the dorsal root produced EPSPs with relatively long latencies, suggesting mediation by a polysynaptic pathway. EPSPs with short latencies were evoked by stimulation of the medulla oblongata, indicating the presence of a monosynaptic excitatory connection. Action potentials, recorded from peripheral nerve after stimulation of the medulla oblongata, were facilitated by conditioning volleys via ventral roots. This facilitation was blocked by dihydro-beta-erythroidine hydrobromide and atropine sulphate, indicating the cholinergic nature of the EPSP of ventral root origin. The conduction velocities of motor axons and of the ventral roots fibers responsible for production of EPSPs were about the same. The EPSP of ventral root origin had a slower rising time course and lesser sensitivity to shifts of membrane potential than the EPSP of medulla oblongata origin, suggesting that the sites of generation of the former EPSP were on the peripheral dendrites. From the above results, it was concluded that the EPSP of ventral root origin was mediated by recurrent axon collaterals of motoneurons.

Action Potentials↗

Glycine neurotransmitter transporters: an update.

Glycine accomplishes several functions as a transmitter in the central nervous system (CNS). As an inhibitory neurotransmitter, it participates in the processing of motor and sensory information that permits movement, vision, and audition. This action of glycine is mediated by the strychnine-sensitive glycine receptor, whose activation produces inhibitory post-synaptic potentials. In some areas of the CNS, glycine seems to be co-released with GABA, the main inhibitory amino acid neurotransmitter. In addition, glycine modulates excitatory neurotransmission by potentiating the action of glutamate at N-methyl-D-aspartate (NMDA) receptors. It is believed that the termination of the different synaptic actions of glycine is produced by rapid re-uptake through two sodium-and-chloride-coupled transporters, GLYT1 and GLYT2, located in the plasma membrane of glial cells or pre-synaptic terminals, respectively. Glycine transporters may become major targets for therapeutic of pathological alterations in synaptic function. This article reviews recent progress on the study of the molecular heterogeneity, localization, function, structure, regulation and pharmacology of the glycine transporter proteins.

Amino Acid Transport Systems, Neutral↗

[Synaptic action of vestibular nerve afferents on reticular neurons of the isolated medulla oblongata of the goldfish].

Excitatory post-synaptic potentials (EPSPs) were evoked in reticular neurons of isolated goldfish brian stem by stimulation of the ipsilateral vestibular nerve or by direct intra-axonal activation of single vestibular fibres. Composite EPSPs displayed usually two components. Ca2+-deficient, Mn2+-containing solution abolished the delayed chemical components of EPSPs, but did not affect the short latency responses suggesting dual (electrical-chemical) mode of transmission between some vestibular afferents and reticular neurons.

Animals↗

[Analysis of factors limiting the applications of binomial statistics in studies of mediator secretion at neuromuscular synapses].

Binomial parameters of transmitter secretion were calculated on the basis of synaptic potential analyses at frog sartorius muscle. In some synapses negative values of parameter of the p were found. These negative values often occurred at low concentrations of Ca2+ ions and small amplitude of miniature end plate potentials. The obtained data were interpreted in terms of a model which presupposes the heterogeneity of the quantum transmitter release at different sites. A computer imitation of transmitter secretion showed that the appearance of the negative p parameter values and erroneous n estimations in the experiment are related with the shape of the distribution of quantum transmitter release probability and the amplitude of miniature end plate potentials.

Animals↗

[Receptive field plasticity of a polyfunctional neuron in mollusks].

Plastic properties of the receptive field of the identified LPa3 neurone were studied on a semi-intact snail preparation. Mantle area representing part of the vast receptive field of the LPa3 neurone of the left parietal ganglion consists of three zones. The right part of the mantle is innervated by the right pallial and anal nerves; the former transmits to the CNS the short latency rapidly extinguishing component of the synaptic response, while the anal nerve--the slowly extinguishing long latency component. The left part of the mantle is innervated by the LPa3 neurone through the left pallial nerve which transmits the multi-component slowly extinguishing excitatory synaptic potential. The pneumostome area is innervated by the right pallial and anal nerves which transmit a non-extinguishing response. Thus, different areas of the mantle are linked to the LPa3 neurone by a system of parallel channels differing by their plastic properties.

Action Potentials↗

Interaction between mossy fibre and climbing fibre responses in Purkinje cells.

A major problem in cerebellar physiology relates to the manner in which information from mossy fibres and climbing fibres is integrated in the efferent neurones of the cerebellar cortex, the Purkinje cells. Recent findings by Ekerot and Oscarsson [8] and by Ito, Sakurai and Tongroach [13] indicate that the important interaction between mossy fibres and climbing fibres takes place in the dendrites rather than in the Purkinje cell somata. It has been demonstrated that impulses in climbing fibres evoke not only the so-called "complex spikes" in the somata but also plateau-like depolarizations in the distal dendrites which may have durations of hundreds of ms. The plateau potentials are presumably produced by a voltage dependent calcium conductance increase which, under physiological conditions, is triggered off exclusively by the large synaptic potentials generated by climbing fibre impulses. The spread of the plateau potentials to the distal dendrites and their long duration would facilitate spatial and temporal interaction between the mossy fibre and climbing fibre inputs. The findings suggest two kinds of interaction. (1) The duration of the plateau potentials in individual dendritic branches is modulated by the local mossy fibre/parallel fibre input. Thus, it might be postulated that dendritic branches act as independent integrators of mossy fibre and climbing fibre inputs. (2) It has been suggested that interaction between mossy fibres and climbing fibres forms the basis of learning processes in the cerebellum mediated through plastic changes in the synapses between parallel fibres and Purkinje cell dendrites. The plateau potentials would influence parallel fibre synapses by increasing the intradendritic calcium which, in turn, would lastingly depress the sensitivity of the postsynaptic receptors of the parallel fibre synapses that are activated in conjunction with the climbing fibres.

Animals↗

A lectin, peanut agglutinin, as a probe for the extracellular matrix in living neuromuscular junctions.

The extracellular matrix plays important roles in the differentiation of synapses. To identify molecules concentrated specifically in the synaptic extracellular matrix, fluorescently-labelled lectins were applied to neuromuscular junctions. A lectin, peanut agglutinin (PNA), stains the neuromuscular region selectively and irreversibly (up to at least 3 weeks in situ), outlining the periphery of the nerve terminal arborization in the frog. Snake neuromuscular junctions also stain intensely with fluorescent PNA, while mouse diaphragm staining is faint. At the electron microscopic level, the reaction products of horseradish peroxidase-conjugated PNA are found primarily in the extracellular matrix flanking Schwann cells in the frog endplate regions. Fluorescently labelled PNA does not affect synaptic potentials and can serve as a simple stain for correlating functional studies of living neuromuscular junctions. Moreover, it can be combined with a presynaptic dye to observe nerve terminals and synaptic extracellular matrix in the same junctions in situ. This report reveals the existence of synapse-specific carbohydrates associated with Schwann cell extracellular matrix in the frog neuromuscular junction. The specific binding and its physiological compatibility make PNA a useful probe for further investigation of synaptic differentiation, plasticity and maintenance.

Animals↗

Slow postsynaptic potentials in neurones of submucous plexus of guinea-pig caecum and their mimicry by noradrenaline and various peptides.

Intracellular recordings of membrane potential and membrane currents were made from neurones in the submucous plexus of the guinea-pig caecum in vitro. Fast and slow excitatory postsynaptic potentials and slow inhibitory postsynaptic potentials were recorded from the majority of neurones following focal stimulation of presynaptic fibres in the plexus. The slow inhibitory postsynaptic potential was associated with an increase in membrane conductance and reversed its polarity at -90 mV; it was reversibly blocked by yohimbine. The slow excitatory postsynaptic potential and its underlying current was associated with a decrease in membrane conductance. Two kinds of voltage-dependence both of the slow excitatory postsynaptic potential and current were observed; in 80% of cells, the excitatory postsynaptic potential and current became smaller with membrane hyperpolarization and reversed polarity at -90 mV (reversing type) but in 20% of cells both the excitatory postsynaptic potential and current simply disappeared when the membrane potential reached -70 mV (non-reversing type). The effects of acetylcholine, adenosine 5'-triphosphate, bombesin, 5-hydroxytryptamine, neurotensin, noradrenaline, substance P and vasoactive intestinal polypeptide were examined. The only substance which mimicked the slow inhibitory postsynaptic potential was noradrenaline; brief applications of noradrenaline caused hyperpolarizations which had the same time-course, reversal potential and sensitivity to yohimbine as the slow inhibitory postsynaptic potential. The non-reversing type of slow excitatory postsynaptic potential was mimicked only by adenosine 5'-triphosphate. The reversing type of slow excitatory postsynaptic potential was mimicked by bombesin, neurotensin, substance P and vasoactive intestinal polypeptide. 5-Hydroxytryptamine and vasoactive intestinal polypeptide (in some neurones) caused a depolarization with an increase in membrane conductance. All three synaptic potentials were reversibly depressed by superfusion of noradrenaline but noradrenaline did not affect the potential changes evoked by brief application of exogenous acetylcholine or substance P. It is concluded that, in guinea-pig submucous plexus neurones, the slow inhibitory postsynaptic potential is mediated by noradrenaline and results from a potassium conductance increase.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine Triphosphate↗

Attenuated hippocampal long-term potentiation in basolateral amygdala-lesioned rats.

Possible involvement of the amygdaloid input in long-term potentiation (LTP) in the medial perforant path-dentate gyrus granule cell synapses in vivo was investigated by evaluating the effects of lesions of the amygdaloid nucleus. The dentate gyrus synaptic potential evoked by low-frequency test stimulation did not change following lesions of the basolateral and central amygdala. However, when tetanic stimulation (30 pulses at 60 Hz) was applied 60 min after lesioning of the ipsilateral basolateral amygdala, the magnitude of LTP was significantly attenuated. Since lesions of the ipsilateral central amygdala and the contralateral basolateral amygdala did not affect the dentate gyrus LTP, the attenuation of the dentate gyrus LTP is a specific effect of acute lesions of the ipsilateral basolateral amygdala. The basolateral amygdaloid lesions significantly attenuated both LTP induced by weak (20 pulses at 60 Hz) and strong (100 pulses at 100 Hz) tetanus, indicating that the effect of the lesions does not depend on the strength of tetanus applied to induce LTP. When the ipsilateral basolateral amygdala was destroyed after application of tetanus, it did not affect the established LTP. The attenuation of LTP was also observed after the basolateral amygdala-lesioned rats were allowed a recovery period of 2 weeks. This is the first report providing evidence that the ipsilateral basolateral amygdala modulates hippocampal synaptic plasticity in vivo.

Amygdala↗

Glutamatergic N2v cells are central pattern generator interneurons of the lymnaea feeding system: new model for rhythm generation.

We aimed to show that the paired N2v (N2 ventral) plateauing cells of the buccal ganglia are important central pattern generator (CPG) interneurons of the Lymnaea feeding system. N2v plateauing is phase-locked to the rest of the CPG network in a slow oscillator (SO)-driven fictive feeding rhythm. The phase of the rhythm is reset by artificially evoked N2v bursts, a characteristic of CPG neurons. N2v cells have extensive input and output synaptic connections with the rest of the CPG network and the modulatory SO cell and cerebral giant cells (CGCs). Synaptic input from the protraction phase interneurons N1M (excitatory), N1L (inhibitory), and SO (inhibitory-excitatory) are likely to contribute to a ramp-shaped prepotential that triggers the N2v plateau. The prepotential has a highly complex waveform due to progressive changes in the amplitude of the component synaptic potentials. Most significant is the facilitation of the excitatory component of the SO --> N2v monosynaptic connection. None of the other CPG interneurons has the appropriate input synaptic connections to terminate the N2v plateaus. The modulatory function of acetylcholine (ACh), the transmitter of the SO and N1M/N1Ls, was examined. Focal application of ACh (50-ms pulses) onto the N2v cells reproduced the SO --> N2v biphasic synaptic response but also induced long-term plateauing (20-60 s). N2d cells show no endogenous ability to plateau, but this can be induced by focal applications of ACh. The N2v cells inhibit the N3 tonic (N3t) but not the N3 phasic (N3p) CPG interneurons. The N2v --> N3t inhibitory synaptic connection is important in timing N3t activity. The N3t cells recover from this inhibition and fire during the swallow phase of the feeding pattern. Feedback N2v inhibition to the SO, N1L protraction phase interneurons prevents them firing during the retraction phase of the feeding cycle. The N2v --> N1M synaptic connection was weak and only found in 50% of preparations. A weak N2v --> CGC inhibitory connection prevents the CGCs firing during the rasp (N2) phase of the feeding cycle. These data allowed a new model for the Lymnaea feeding CPG to be proposed. This emphasizes that each of the six types of CPG interneuron has a unique set of synaptic connections, all of which contribute to the generation of a full CPG pattern.

Acetylcholine↗

Potentiation of GABAergic synaptic transmission by AMPA receptors in mouse cerebellar stellate cells: changes during development.

1. The effects of low concentrations of domoate, an agonist at both alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate and kainate receptors (AMPARs and KARs, respectively), were investigated in stellate cells in slices of mouse cerebellum at two developmental stages (postnatal day (PN) 11-13 and PN21-25). 2. Low concentrations of domoate enhanced the frequency of miniature IPSCs (mIPSCs) recorded in the presence of tetrodotoxin (TTX) at PN11-13 but not at PN21-25. 3. The effects of low concentrations of domoate on synaptic activity were probably mediated by the activation of AMPARs and not KARs, since they were blocked by GYKI 53655 (LY300168), a selective AMPAR antagonist. 4. Domoate increased mIPSC frequency in part by activation of presynaptic voltage-dependent Ca2+ channels since potentiation was reduced by 60 % in the presence of Cd2+. AMPARs in stellate cells were found to be permeable to Ca2+. The residual potentiation in the presence of Cd2+ could thus be due to a direct entry of Ca2+ through AMPAR channels. 5. In the presence of TTX, potentiation of synaptic activity by focal application of domoate was not restricted to the region of the cell body, but was observed within distances of 120 micro(m). These experiments also revealed a strong spatial correlation between the location of the presynaptic effects of domoate and the activation of postsynaptic AMPARs. 6. Our data show a developmentally regulated presynaptic potentiation of synaptic transmission between cerebellar interneurones mediated by AMPARs. We discuss the possibility that the developmental switch could be due to a shift in the localization of AMPARs from the axonal to the somato-dendritic compartment.

Age Factors↗