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Neurotransmitter release requires high energy demands, making the nerve terminals metabolically fragile and susceptible to oxidative stress. ATP-sensitive potassium (KMicroelectrode recordings of end plate potentials at the distal and proximal compartments of nerve terminals as well as fluorescent techniques were used.Activation of KK

Nerve terminals contain numerous synaptic vesicles (SVs) whose exo-endocytic cycling maintains neurotransmitter release. SVs may have different properties, thereby constituting separate pools. However, behavior of SV pools remains elusive in many synapses. To fill this gap, we studied the functioning of SV pools at both low- and higher-frequency stimulations utilizing microelectrode recording and dual-labeling of SVs with FM-dyes at the mice motor nerve terminals. It was found that higher-frequency stimulation caused exocytosis of different kinds of SVs. One type of SVs contributed to exocytosis exclusively at intense activities and their exocytotic rate was depended on the order in which these SVs were recovered by endocytosis. Another type of SVs can sustain the release in response to both low- and higher-frequency stimulations, but increasing activity did not lead to enhanced exocytotic rate of these SVs. In addition, depression of neurotransmitter release induced by 20 Hz stimulation occurred independent on previous episode of 10 Hz activity. We suggest that during prolonged stimulation at least two SV pools can operate. One termed "house-keeping" that would be active at different frequencies and the other termed "plug-in" that would respond to increasing activity.

L-type Ca

Adrenoceptor activators and blockers are widely used clinically for the treatment of cardiovascular and pulmonary disorders. More recently, adrenergic agents have also been used to treat neurodegenerative diseases. Recent studies indicate a location of sympathetic varicosities in close proximity to neuromuscular junctions. The pressing question is whether there could be any effects of endo- or exogenous catecholamines on cholinergic neuromuscular transmission. It was shown that the pharmacological stimulation of adrenoceptors, as well as sympathectomy, can affect both acetylcholine release from motor nerve terminals and the functioning of postsynaptic acetylcholine receptors. In this review, we discuss the recent data regarding the effects of adrenergic drugs on neurotransmission at the neuromuscular junction. The elucidation of the molecular mechanisms by which the clinically relevant adrenomimetics and adrenoblockers regulate quantal acetylcholine release from the presynaptic nerve terminals and postsynaptic sensitivity may help in the design of highly effective and well-tolerated sympathomimetics for treating a number of neurodegenerative diseases accompanied by synaptic defects.

Cd2+ is one of the most widespread environmental pollutants and its accumulation in central and peripheral nervous systems leads to neurotoxicity as well as aggravation of common neurodegenerative diseases. Mechanism of the Cd2+ toxicity is far from being resolved. Here, using microelectrode recordings of postsynaptic responses and fluorescent redox indicators we studied the effect of Cd2+ in the submicromolar range on timing of neurotransmitter release and oxidative status in two functionally different compartments of the same frog motor nerve terminal. Cd2+ (0.1–1 μM) acting as typical voltage-gated Ca2+channel (VGCC) antagonist decreased neurotransmitter release in both distal and proximal parts of the nerve terminal, but in contrast to the VGCC blockers Cd2+(0.1–0.5 μM) desynchronized the release selectively in the distal region. The latter action of Cd2+ was completely prevented by inhibitor of NADPH-oxidase and antioxidants, including mitochondrial specific, as well as redox-sensitive TRPV1 channel blocker. Cd2+ markedly increased levels of mitochondrial reactive oxygen species (ROS) in both the distal and proximal compartments of the nerve terminal, which was associated with lipid peroxidation mainly in the distal region. Zn2+, whose transport systems translocate Cd2+, markedly enhanced the effects of Cd2+ on both the mitochondrial ROS levels and timing of neurotransmitter release. Furthermore, in the presence of Zn2+ ions, Cd2+ also desynchronized the neurotransmitter release in the proximal region. Thus, in synapses Cd2+ at very low concentrations can increase mitochondrial ROS, lipid peroxidation and disturb the timing of neurotransmitter release via a ROS/TRPV-dependent mechanism. Desynchronization of neurotransmitter release and synaptic oxidative stress could be early events in Cd2+ neurotoxicity.

At neuromuscular junctions, ATP inhibits both the evoked and spontaneous acetylcholine release and inward calcium current operating via presynaptic P2Y receptors. It was shown in the experiments with the frog neuromuscular synapse using specific calcium-sensitive dye Oregon Green Bapta 1 that exogenous ATP reduces the amplitude of calcium transient, which reflects the changes in the entry of calcium ions in response to the nerve pulse. The depressing effect of ATP on the transient was prevented by suramin, the blocker of P2 receptors. Nitrendipine, a specific blocker of L-type calcium channels, per se decreased the calcium transient amplitude and significantly attenuated the effect of ATP on the calcium signal. Contrariwise, the preliminary application of ATP to the neuromuscular junction completely eliminated the depressing effect of nitrendipine on the calcium response. The obtained data suggest that an essential component in the inhibitory action of ATP on the calcium transient amplitude is provided by reduction of the entry of calcium ions into a frog nerve ending via L-type voltage-gated calcium channels.