Tetanic Depression Research Articles

Соотношение возбудительных и тормозных синаптических процессов в антиноцицептивных ядрах PAG и RMG в их реципрокных взаимоотношениях в интактном препарате с воздействием гидрокортизона

Microelectrophysiological studies were carried out on 12 intact Albino rats (250±30 g) with hydrocortisone injection, compared with the norm. A software mathematical analysis of a single spike activity of 282 neurons was carried out. The spike activity of single PAG and RMG neurons was recorded extracellularly in separate series of experiments, with successive involvement of each of them as stimulated and recorded. When assessing the relative degree severity of depressor and excitatory post-stimulus effects, using the example of diagrams of the average spike frequency in PAG at HFS RMG under hydrocortisone exposure in normal conditions, both in the depressor and excitatory sequences, in comparison with those without hydrocortisone, no significant changes were detected, but there was a greater severity of these values in the norm without hydrocortisone, in particular, preceding excitatory effects, which indicates not only the harmlessness of the indicated dose of hydrocortisone, but also a certain improvement in the normal status. In RMG neurons on HFS PAG, both in the norm without and under hydrocortisone exposure, a protective increase in tetanic depression was revealed in both sequences, without excess of poststimulus excitability. In general, for both PAG and RMG, a greater severity of pre- and post-stimulus frequency, preceding and accompanying excitatory effects in the norm without hydrocortisone, and a greater balance of depressor and excitatory post-stimulus manifestations of activity under hydrocortisone exposure than in the norm were revealed, which indicates the safety and even the benefit of that dose of hydrocortisone.

Plasticity in Motoneurons Following Spinal Cord Injury in Fructose-induced Diabetic Rats.

Spinal cord injury (SCI) causes motor impairment and the proper excitation/inhibition balance in motoneurons is important for recovery. Diabetes mellitus impairs regenerative capacity following SCI. The purpose of this study was to assess the short-term plasticity (STP) of lumbar spinal cord motoneurons in conditions of (1) lateral hemisection (SCI), (2) fructose-induced diabetes (D), and (3) diabetes associated with hemisection (D + SCI). We show that in the cases of SCI, D, and D + SCI, the ratio of percentage share of excitatory and inhibitory combinations of motoneurons responses to high-frequence stimulation of sciatic nerve is multidirectional. In the SCI and D + SCI groups, the cumulative changes in generalized baseline frequencies decreased significantly. When we compared the cumulative changes in the intensity of excitatory and inhibitory responses relative to baseline during high-frequency stimulation (tetanization epoch), we found that there was a significant intensification in tetanic depression in the D + SCI groups versus SCI, as well as an intensification in tetanic potentiation in the D + SCI vs. D and D + SCI vs. SCI groups. Thus, in conditions of traumatic and/or metabolic pathology, the distinct synaptic inputs exhibit opposing plasticity for homeostatic control of neurotransmission and these integral changes most likely shape postsynaptic STP in the spinal motor network.

Homeostatic plasticity induced by increased acetylcholine release at the mouse neuromuscular junction

At the neuromuscular junction (NMJ), changes to the size of the postsynaptic potential induce homeostatic compensation. At the Drosophila NMJ, increased glutamate release causes a compensatory decrease in quantal content, but it is unknown if this mechanism operates at the cholinergic mammalian NMJ. We addressed this question by recording endplate potentials (EPP) and muscle contraction in 3-month and 24-month ChAT-ChR2-EYFP mice that overexpress vesicular acetylcholine transporter and release more acetylcholine per vesicle. At 3 months, the quantal content of EPPs from ChAT-ChR2-EYFP mice were not different from WT controls, however tetanic depression was greater, and quantal size during high-frequency stimulation and the size of the readily releasable pool (RRP) were decreased. At 24 months of age, quantal content was reduced in ChAT-ChR2-EYFP mice, which normalized synaptic depression despite smaller RRP. The effect of pancuronium on indirect evoked muscle twitch was not different between groups. These results indicate that an increase in the amount of acetylcholine per vesicle induces two distinct age-dependent homeostatic mechanisms compensating excessive acetylcholine release.

Microelectrophysiological Studies of the Ratio of Excitatory to Inhibitory Synaptic Processes in the Corticonigral Projection in a Model of Parkinson’s

Experiments on 23 white male rats (250 g) analyzed the spike activity of individual neurons in the substantia nigra pars compacta (SNc, 242 neurons, n = 11) and substantia nigra pars reticulata (SNr, 289 neurons, n = 12) during high-frequency stimulation of the primary motor cortex (M1)in normal animals and in animals with a rotenone model of Parkinson’s disease (BP). SNc neurons in the model of PD showed a complete absence of depressor effects induced by stimulation, though tetanic potentiation was accompanied by posttetanic potentiation and depression at levels 1.65 and 2.02 times greater than in normal animals. In SNr neurons in normal animals, tetanic potentiation, accompanied by post-tetanic potentiation and depression, was 2.37 times greater than tetanic depression, while in the model of PD the levels of both depressor and excitatory activity induced by stimulation were below normal. Spike activity frequency in SNc and SNr neurons preceding and accompanying stimulation was significantly greater than normal in the model of PD. This is evidence for excitotoxicity accompanying neurodegenerative damage, which is completed by neuron apoptosis and death. In SNr neurons, both depressor and excitatory reactions accompanying stimulation were markedly dominant over those in SNc neurons, which is evidence for more extensive cortical projections to the SNr. Furthermore, SNc neurons demonstrated greater susceptibility to pathological changes due to poststimulus depressor effects than SNr neurons, with formation of more marked excitatory effects, which is evidence of a greater involvement of the SNc in PD. In the model of PD, lacking stimulation-induced depressor effects and more marked excitatory effects in SNc neurons, SNr neurons retained their depressor reactions and relatively decreased excitatory reactions, which is evidence of a lower level of susceptibility of SNr neurons to excitotoxicity, extreme increases in the excitability of surviving neurons compensating for the lack of excitation of dead cells.

Микроэлектрофизиологическое изучение соотношения возбудительных и тормозных синаптических процессов в кортико-нигральной проекции на модели болезни Паркинсона

The impulse activity of single neurons in substantia nigra pars compacta - SNc (242 neurons, n=11) and substantia nigra pars reticulate - SNr (289 neurons, n=12) under high frequency stimulation of primary motor cortex (M1) was analyzed on 23 white male rats in control and rotenone model of Parkinson’s disease (PD). In SNc neurons in PD model there was no stimulation-induced depression, however, the posttetatic potentiation and depression following the tetanic potentiation, were 1.65 and 2.02 times higher than in control. In control SNr neurons the tetanic potentiation followed by the posttetanic potentiation and depression was 2.37 times higher than the tetanic depression, whereas in the PD model both depression and potentiation levels were subnormal. In PD model, both SNc and SNr neurons exhibited significantly higher spike frequency before and during the stimulation in comparison with control. This indicates the presence of the excitotoxicity accompanied by the neurodegenerative damage followed by the apoptosis and the neuronal death. In SNr neurons both depression and potentiation after the stimulation were much more prominent than in SNc neurons, which indicates that the cortical projections on the SNr are more pronounced. Moreover, the SNc neurons demonstrated higher succeptibility to the pathological changes in the poststimulus depression, leading to the more pronounced potentiation in those neurons in comparison with the SNr neurons, which indicates the more prominent involvement of the former in the PD pathogenesis. In the PD model, with no stimulation-induced depression and more prominent potentiation in the SNc neurons, the SNr neurons maintain the depression and show relatively lowered potentiation, which indicates their lower succeptibility to the excitotoxicity, and the excessive excitability of the remaining neurons as a compensation for the dead ones.

Tetanic Facilitation of Neuromuscular Transmission by Adenosine A2A and Muscarinic M1 Receptors is Dependent on the Uptake of Choline via High-Affinity Transporters

Background/Aims: In this study, we evaluated the functional impact of facilitatory presynaptic adenosine A_2A and muscarinic M₁ receptors in the recovery of neuromuscular tetanic depression caused by the blockage of high-affinity choline transporter (HChT) by hemicholinium-3 (HC-3), a condition that mimics a myasthenia-like condition. Methods: Rat diaphragm preparations were indirectly stimulated via the phrenic nerve trunk with 50-Hz frequency trains, each consisting of 500–750 supramaximal intensity pulses. The tension at the beginning (A) and at the end (B) of the tetanus was recorded and the ratio (R) B/A calculated. Results: Activation of A_2A and M₁ receptors with CGS21680 (CGS; 2 nmol/L) and McN-A-343c (McN; 3 μmol/L) increased R values. Similar facilitatory effects were obtained with forskolin (FSK; 3 μmol/L) and phorbol 12-myristate 13-acetate (PMA; 10 μmol/L), which activate adenylate cyclase and protein kinase C respectively. HC-3 (4 μmol/L) decreased transmitter exocytosis measured by real-time videomicroscopy with the FM4-64 fluorescent dye and prevented the facilitation of neuromuscular transmission caused by CGS, McN, and FSK, with a minor effect on PMA. The acetylcholinesterase inhibitor, neostigmine (NEO; 0.5 μmol/L), also decreased transmitter exocytosis. The paradoxical neuromuscular tetanic fade caused by NEO (0.5 μmol/L) was also prevented by HC-3 (4 μmol/L) and might result from the rundown of the positive feedback mechanism operated by neuronal nicotinic receptors (blocked by hexamethonium, 120 μmol/L). Conclusion: Data suggest that the recovery of tetanic neuromuscular facilitation by adenosine A_2A and M₁ receptors is highly dependent on HChT activity and may be weakened in myasthenic patients when HChT is inoperative.

Neuroprotective Effects of Macrovipera lebetina Snake Venom in the Model of Alzheimer’s Disease

Pathological features of Alzheimer’s disease (AD) include accumulation and deposition of β-amyloid (Aβ) in the brain, activation of astrocytes and microglia, and disruption of cholinergic neurotransmission. In our experiments on rats, a model of AD was created by intracerebroventricular (i.c.v.) injections of Aβ25–35 amyloid. In another animal group, this was combined with intramuscular (i.m.) injections of small doses of Macrovipera lebetina (ML) snake venom (50 μl of 5% solution of an LD50 dose per animal seven times with one-day-long intervals). In the AD model, the most vulnerable neurons were found in the hippocampal fields CA1 and CA3. The phosphatase activity in the hippocampus of Aβ-injected rats sharply dropped. Systemic administration of small doses of ML venom induced positive changes in the structural characteristics of hippocampal neurons, increased the density of neurons in the above fields, normalized metabolism, and intensified Ca2+-dependent phosphorylation. Under the action of ML venom, the proportion of responses of hippocampal pyramidal neurons after high-frequency tetanic stimulation of the ipsilateral entorhinal cortex in the form of tetanic depression–posttetanic potentiation increased, and an overall increase in the firing rate of hippocampal neurons was observed. The intensity of free radical processes in some tissues of Aβ-affected animals became much lower under the action of ML venom. Thus, small doses of this venom manifest clear neuroprotective effects in the rat AD model.

Central activation, metabolites, and calcium handling during fatigue with repeated maximal isometric contractions in human muscle.

To determine the roles of calcium (Ca2+) handling by sarcoplasmic reticulum (SR) and central activation impairment (i.e., central fatigue) during fatigue with repeated maximal voluntary isometric contractions (MVC) in human muscles. Contractile performance was assessed during 3 min of repeated MVCs (7-s contraction, 3-s rest, n=17). In ten participants, in vitro SR Ca2+-handling, metabolites, and fibre-type composition were quantified in biopsy samples from quadriceps muscle, along with plasma venous [K+]. In 11 participants, central fatigue was compared using tetanic stimulation superimposed on MVC in quadriceps and adductor pollicis muscles. The decline of peak MVC force with fatigue was similar for both muscles. Fatigue resistance correlated directly with % type I fibre area in quadriceps (r=0.77, P=0.009). The maximal rate of ryanodine-induced Ca2+-release and Ca2+-uptake fell by 31±26 and 28±13%, respectively. The tetanic force depression was correlated with the combined reduction of ATP and PCr, and increase of lactate (r=0.77, P=0.009). Plasma venous [K+] increased from 4.0±0.3 to 5.4±0.8mM over 1-3-min exercise. Central fatigue occurred during the early contractions in the quadriceps in 7 out of 17 participants (central activation ratio fell from 0.98±0.05 to 0.86±0.11 at 1min), but dwindled at exercise cessation. Central fatigue was seldom apparent in adductor pollicis. Fatigue with repeated MVC in human limb muscles mainly involves peripheral aspects which include impaired SR Ca2+-handling and we speculate that anaerobic metabolite changes are involved. A faster early force loss in quadriceps muscle with some participants is attributed to central fatigue.

Ryanodine and inositol triphosphate receptors modulate facilitation and tetanic depression at the frog neuromuscular junction.

Short-term plasticity of synaptic function is an important physiological control of transmitter release. Short-term plasticity can be regulated by intracellular calcium released by ryanodine and inositol triphosphate (IP3) receptors, but the role of these receptors at the neuromuscular junction is understood incompletely. We measured short-term plasticity of evoked endplate potential (EPP) amplitudes from frog neuromuscular junctions treated with ryanodine, 2-aminoethoxydiphenylborane (2-APB), or 1-[6-[[(17β)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1H-pyrrole-2,5-dione (U- 73122). Ryanodine decreases paired-pulse facilitation for intervals <20 ms and markedly decreases tetanic depression. Treatment with 2-APB reduces EPP amplitude, increases paired-pulse facilitation for intervals of <20 ms, and significantly reduces tetanic depression. U-73122 decreases EPP amplitude and decreases paired-pulse depression for intervals <20 ms. Ryanodine, IP3 receptors, and phospholipase C modulate short-term plasticity of transmitter release at the neuromuscular junction. These results suggest possible targets for improving the safety factor of neuromuscular transmission during repetitive activity of the neuromuscular junction.

The tetanic depression in fast motor units of mammalian skeletal muscle can be evoked by lengthening of one initial interpulse interval

A lower than expected tetanic force (the tetanic depression) is regularly observed in fast motor units (MUs) when a higher stimulation frequency immediately follows a lower one. The aim of the present study was to determine whether prolongation of only the first interpulse interval (IPI) resulted in tetanic depression. The experiments were carried out on fast MUs of the medial gastrocnemius muscle in cats and rats. The tetanic depression was measured in each case as the force decrease of a tetanus with one IPI prolonged in relation to the tetanic force at the respective constant stimulation frequency. Force depression was observed in all cases studied and was considerably greater in cats. For cats, the mean values of force depression amounted to 28.64% for FR and 10.86% for FF MUs whereas for rats 9.30 and 7.21% for FR and FF motor units, respectively. Since the phenomenon of tetanic depression in mammalian muscle is commonly observed even after a change in only the initial interpulse interval within a stimulation pattern, it can effectively influence processes of force regulation during voluntary activity of a muscle, when motoneurones progressively increase the firing rate.

Tetanic failure due to decreased endogenous adenosine A2A tonus operating neuronal Cav1 (L-type) influx in Myasthenia gravis

In healthy motor endplates, tetanic depression is overcome by tonic adenosine A(2A) -receptor-mediated facilitation of transmitter release. The A(2A) receptor operates a coordinated shift from fast-desensitizing Ca(v) 2.1 (P/Q) calcium influx to long-lasting Ca(V) 1 (L) channels on motor nerve terminals. This study aimed at investigating whether A(2A) receptors-operated Ca(2+) influx via Ca(V) 1 (L)-type channels contribute to sustain acetylcholine release evoked by 50 Hz-bursts in toxin-induced Myasthenia gravis (TIMG) rats. In contrast to control animals, inhibition of [(3) H]acetylcholine (ACh) release by the Ca(V) 2.1 (P/Q) channel blocker, ω-Agatoxin IVA (100 nM), in TIMG rats had a higher magnitude than that observed with the Ca(V) 1 (L) channel blocker, nifedipine (1 μM). Adenosine deaminase (0.5 U/mL) and the A(2A) receptor antagonist, ZM 241385 (50 nM), decreased [(3) H]ACh release by a similar amount in control rats, but their effects were smaller in magnitude in myasthenic animals. The adenosine precursor, AMP (100 μM), increased (~40%) ACh release in both control and TIMG animals. Blockade of A(2A) , but not of A(1) , receptors prevented AMP-induced facilitation of transmitter release; nifedipine (1 μM) mimicked the effect of the A(2A) receptor antagonist. Video-microscopy studies designed to measure real-time transmitter exocytosis using the FM4-64 fluorescent dye fully supported radiochemical data. Thus, impairment of the adaptive shift from Ca(V) 2.1 (P/Q) to Ca(V) 1 (L) channels may contribute to tetanic failure in myasthenic rats. This parallels the reduction of adenosine A(2A) receptor tonus in TIMG animals, which might be restored by exogenous application of AMP.

Quantal release of acetylcholine in mice with reduced levels of the vesicular acetylcholine transporter

Mammalian motor nerve terminals contain hundreds of thousands of synaptic vesicles, but only a fraction of these vesicles is immediately available for release, the remainder forming a reserve pool. The supply of vesicles is replenished through endocytosis, and newly formed vesicles are refilled with acetylcholine through a process that depends on the vesicular acetylcholine transporter (VAChT). During expression of short-term plasticity, quantal release can be increased, but it is unknown whether this reflects enhanced recruitment of vesicles from the reserve pool or rapid recycling. We examined spontaneous and evoked release of acetylcholine at endplates from genetically modified VAChT KD(HOM) mice that express approximately 30% of the normal level of VAChT to determine steps rate-limited by synaptic vesicle filling. Quantal content and quantal size were reduced in VAChT KD(HOM) mice compared with wild-type controls. Although high-frequency stimulation did not reduce quantal size further, the post-tetanic increase in end-plate potential amplitude or MEPP frequency was significantly smaller in VAChT KD(HOM) mice. This was the case even when tetanic depression was eliminated using an extracellular solution containing reduced Ca(2+) and raised Mg(2+). These results reveal the dependence of short-term plasticity on the level of VAChT expression and efficient synaptic vesicle filling.

Tetanic depression and catch-like effect in fast motor units of the rat medial gastrocnemius at linearly increasing and decreasing stimulation frequencies

The forces developed by fast resistant (FR) and fast fatigable (FF) motor units of the rat medial gastrocnemius during trains of electrical stimuli at linearly increasing or decreasing frequency were measured at the instantaneous frequency of 60 Hz and compared with the force evoked at a constant 60 Hz. In both motor unit types, the mean forces during stimulation at increasing frequency were depressed by 17%, while those recorded during stimulation at decreasing frequency were increased by 15% (FR) or 10% (FF) compared to values observed during constant-rate stimulation. During trains of stimuli at an increasing rate, the instantaneous frequency necessary to induce a force comparable to that produced at constant 60 Hz stimulation in FR and FF units was 84 and 88 Hz, respectively; whereas for the same units during stimulation at a decreasing rate these values were 45 and 47 Hz, respectively. When the stimulation frequency was increased up to 60 Hz and then held at this level, the force of both motor unit types was depressed by approximately 6% compared to 60 Hz constant-rate stimulation. From the available data it may be concluded that the phenomenon of tetanic depression is able to limit the development of force when the motoneuronal firing rate increases.

The role of spontaneous activity in development of the endbulb of Held synapse

In the mouse brainstem cochlear nucleus, the auditory nerve to bushy cell synapse (endbulb of Held) is specialised for rapid, high-fidelity transmission. Development of this synapse is modulated by auditory nerve activity. Here we investigate the role of spontaneous auditory nerve activity in synaptic transmission using deafness ( dn/ dn) mutant mice that have abnormal hair cells and lack spontaneous auditory nerve activity. Evoked and miniature alpha amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptor-mediated excitatory post-synaptic currents (eEPSCs, mEPSCs) were compared in deafness and normal mice before the age of hearing onset (postnatal day 7–11: P7–11) using variance-mean, miniature event and tetanic depression analyses. Amplitudes were significantly greater in deafness mice for eEPSCs (2.1-fold), mEPSCs (1.4-fold) and quantal amplitudes (1.5-fold). eEPSCs in deafness mice decayed more rapidly with increasing age, indicating an input-independent transition in post-synaptic AMPA receptor properties. A comparison of normal mice before and after the onset of hearing showed a change in synaptic parameters with an increase in eEPSC (1.7-fold), mEPSC (1.6-fold) and quantal amplitude (1.7-fold) after hearing onset while release probability remained constant (0.5). Overall, the results in deafness mice suggest that synaptic strength is altered in the absence of spontaneous auditory nerve activity.

Development of a Robust Central Auditory Synapse in Congenital Deafness

Within the medial nucleus of the trapezoid body (MNTB) in the auditory brain stem, there is a large central synapse known as the calyx of Held, which mediates high-fidelity glutamatergic transmission. We investigated the effects of congenital deafness on the development of pre- and postsynaptic parameters of synaptic strength at the calyx of Held. Whole cell recordings of evoked excitatory postsynaptic currents (EPSCs) and immunohistochemistry of GluR1-4 subunits were performed using brain stem slices from congenitally deaf or hearing mice at postnatal days P5 and P12. In both hearing and deaf mice there was a similar developmental decrease in the NMDA component of the evoked EPSC. There was a concurrent increase in release probability and number of release sites, contributing to a fivefold increase in evoked AMPA-mediated EPSC amplitude. The increase in release probability is opposite to that found in previous studies at the calyx of Held in the rat. There was also a seven- to eightfold increase in the size of the readily releasable pool of vesicles and a decrease in tetanic depression. The postsynaptic glutamate receptor subunits were similarly developmentally regulated and unaffected by deafness. GluR1 and 4 dominated at both ages. There was a decrease in expression of GluR1-3 from P5 to P12 and a shift from GluR2 to GluR3, indicating that AMPA receptor complexes at P12 are predominantly calcium-permeable. These results demonstrate that early development at this robust synapse proceeds normally with congenital deafness, suggesting that auditory nerve activity does not affect the development of synaptic strength at the calyx of Held.

Tetanic depression is overcome by tonic adenosine A(2A) receptor facilitation of L-type Ca(2+) influx into rat motor nerve terminals.

Motor nerve terminals possess multiple voltage-sensitive calcium channels operating acetylcholine (ACh) release. In this study, we investigated whether facilitation of neuromuscular transmission by adenosine generated during neuronal firing was operated by Ca(2+) influx via 'prevalent' P-type or via the recruitment of 'silent' L-type channels. The release of [(3)H]ACh from rat phrenic nerve endings decreased upon increasing the stimulation frequency of the trains (750 pulses) from 5 Hz (83 +/- 4 x 10(3) disintegrations per minute per gram (d.p.m. g(-1)); n = 11) to 50 Hz (30 +/- 3 x 10(3) d.p.m. g(-1); n = 5). The P-type Ca(2+) channel blocker, omega-agatoxin IVA (100 nm) reduced (by 40 +/- 10%; n = 6) the release of [(3)H]ACh evoked by 50-Hz trains, while nifedipine (1 microM, an L-type blocker) was inactive. Tetanic depression was overcome (88 +/- 6 x 10(3) d.p.m. g(-1); n = 12) by stimulating the phrenic nerve with 50-Hz bursts (five bursts of 150 pulses, 20 s interburst interval). In these conditions, omega-agatoxin IVA (100 nM) failed to affect transmitter release, but nifedipine (1 microM) decreased [(3)H]ACh release by 21 +/- 7% (n = 4). Inactivation of endogenous adenosine with adenosine deaminase (ADA, 0.5 U ml(-1)) reduced (by 54 +/- 8%, n = 5) the release of [(3)H]ACh evoked with 50-Hz bursts. This effect was opposite to the excitatory actions of adenosine (0.5 mm), S-(p-nitrobenzyl)-6-thioinosine (5 microM, an adenosine uptake blocker) and CGS 21680C (3 nM, a selective A(2A) receptor agonist); as the A(1) receptor agonist R-N(6)-phenylisopropyl adenosine (R-PIA, 300 nM) failed to affect the release of [(3)H]ACh, the results indicate that adenosine generated during 50-Hz bursts exerts an A(2A)-receptor-mediated tonus. The effects of ADA (0.5 U ml(-1)) and CGS 21680C (3 nm) were prevented by nifedipine (1 microM). Blocking tonic A(2A) receptor activation, with ADA (0.5 U ml(-1)) or 3,7-dimethyl-1-propargyl xanthine (10 microM, an A(2A) antagonist), recovered omega-agatoxin IVA (100 nM) inhibition and caused the loss of function of nifedipine (1 microM). Data indicate that, in addition to the predominant P-type Ca(2+) current triggering ACh release during brief tetanic trains, motoneurones possess L-type channels that may be recruited to facilitate transmitter release during high-frequency bursts. The fine-tuning control of Ca(2+) influx through P- or L-type channels is likely to be mediated by endogenous adenosine. Therefore, tonic activation of presynaptic A(2A) receptors operating Ca(2+) influx via L-type channels may contribute to overcome tetanic depression during neuronal firing.

Facilitation and Depression of Neuromuscular Transmission under Conditions of Blockade of Ryanodine Receptors

In our experiments on mice, end-plate currents (EPC) evoked by stimulation of the phrenic nerve were intracellularly recorded in neuromuscular synaptic junctions of the phrenic muscle. We studied the effects of a specific blocker of ryanodine receptors, ryanodine (10 to 20 μM), on the amplitude and time parameters of EPC under conditions of tetanic facilitation and depression of synaptic transmission at frequencies of stimulation of 4 to 200 sec-1. Ryanodine inhibited facilitation at stimulation frequencies of 7 to 70 sec-1 (with maximum effect at 20 sec-1) and accelerated depression. In the presence of ryanodine, an initial rundown of the EPC amplitude in the course of depression of transmission increased at high frequencies of stimulation (50 to 100 sec-1), whereas the EPC amplitude at the plateau level decreased already at low frequencies (4 to 7 sec-1). We concluded that the changes in facilitation and depression resulted from blocking of the presynaptic ryanodine receptors by ryanodine. It seems probable that calcium release from the calcium stores in murine motor terminals is a factor involved in the control of processes of transmitter secretion during short-term rhythmic activation of the junction.

Synaptic transmission in the auditory brainstem of normal and congenitally deaf mice.

The deafness (dn/dn) mutant mouse provides a valuable model of human congenital deafness. We investigated the properties of synaptic transmission in the anteroventral cochlear nucleus (AVCN) of normal and congenitally deaf dn/dn mice. Excitatory postsynaptic currents (EPSCs) were evoked by focal stimulation of single auditory nerve fibres, and measured by whole-cell recordings from neurones in AVCN slices (mean postnatal age = P13). Absolute amplitudes of both AMPA- and NMDA-mediated components of evoked EPSCs were greater (170 %) in deaf versus control animals. Enhanced transmission in deaf mice was due to a presynaptic mechanism. Variance-mean analysis revealed that the probability of transmitter release was significantly greater in deaf (P(r) = 0.8) versus control animals (P(r) = 0.5). Following high frequency stimulation, deaf mice showed a greater depression of evoked EPSCs, and a significant increase in the frequency of delayed-release (asynchronous) miniature EPSCs (aEPSCs) (deaf 100 Hz vs. control 7 Hz). The acetoxymethyl ester of EGTA (EGTA-AM) blocked the increase in miniature aEPSCs and returned tetanic depression to control values. In deaf mice, reduction of mean P(r) using cadmium caused an expected increase in paired-pulse ratio (PPR). However, in the same cells, a similar reduction in release by EGTA-AM did not result in a change in PPR, demonstrating that a change may occur in P(r) without a concomitant change in PPR. In many respects, transmission in deaf mice was found to be remarkably similar to control mice, implying that many parameters of synaptic transmission develop normally in these animals. The two significant differences (higher P(r) and asynchronous release in deaf mice) could both be reversed by the addition of EGTA-AM, suggesting that endogenous calcium buffering may be impaired or undeveloped in the presynaptic terminals of the auditory nerve in deaf mice.

Tetanic depression: a phenomenon influencing the production of tension in fast-twitch motor units in rat medial gastrocnemius

The influence of an increase or a decrease in the stimulation frequency on tension development during a tetanus was studied in motor units of the rat medial gastrocnemius muscle. These effects were tested in one tetanus evoked at two frequencies of stimulation, a lower immediately followed by a higher one or the reverse. For all fast motor units it was observed that after the first part of a tetanus at a lower frequency of stimulation the tension of the following part, of the better fused contraction, was depressed. This effect was called a tetanic depression. When the lower stimulation frequency was followed by the higher one, the depression was visible in some motor units only whereas in the remaining units a potentiation of the second part of the tetanus was visible. The tetanic depression was larger in fast resistant than in fast fatigable motor units. In slow motor units tetanic depression was not observed. The tetanic depression is a phenomenon which can influence the production of contractile tension by fast motor units.

Developmental increase in asynchronous GABA release in cultured hippocampal neurons

Developmental changes in GABAergic synaptic transmission were examined in cultured hippocampal neurons using patch-clamp recordings and Ca 2+ imaging. In paired recordings, tetanization of the presynaptic GABAergic neuron with 80 pulses at either 40 or 80 Hz was accompanied by tetanic depression of inhibitory postsynaptic responses. In neurons that had been cultured for more than two weeks, asynchronous inhibitory postsynaptic currents often appeared during the tetanus and continued for several seconds following stimulation. There was little asynchronous activity in neurons that had been cultured for shorter times. However, no age-related changes were observed in the amplitude of single synchronous inhibitory postsynaptic currents, paired-pulse depression or post-tetanic potentiation of inhibitory postsynaptic currents. Following equimolar replacement of extracellular Ca 2+ with strontium ions (Sr 2+), single autaptic inhibitory postsynaptic currents were depressed in amplitude and asynchronous inhibitory postsynaptic currents were present on the decaying phase. Sr 2+-induced asynchronous inhibitory postsynaptic currents showed no dependence on age in culture. Imaging of Ca 2+ in single GABAergic boutons was performed by including Fluo-3 in the patch pipette. During action potential firing induced by stimulating at 80 Hz for 1 s, intracellular calcium [Ca 2+] i increased rapidly in individual boutons. Following the stimulus, [Ca 2+] i decayed back to baseline within 10–15 s. The half-time of decay increased from 1.7±0.2 s at 15 days in vitro to 4.0±0.2 s at 30 days in vitro ( P<0.05), with a developmental profile that closely matched the increase in asynchronous inhibitory postsynaptic currents. We propose that the increase in tetanus-induced asynchronous GABA-release during the first month of synapse maturation in vitro is caused by a slowing of the Ca 2+-clearing mechanisms in the GABAergic boutons. This results in larger and more prolonged elevations of [Ca 2+] i during tetanic stimulation, which leads to enhanced asynchronous transmitter release. We propose that the results of this study demonstrate a potentially important aspect of synapse maturation during development, and also imply that GABA release is up-regulated in conditions of decreased Ca 2+ buffering and clearing.