The release of dopamine in the striatum, nucleus accumbens, and olfactory tubercle of anesthetized rats was evoked by electrical stimulation of the mesolimbic dopaminergic pathway (four pulses at 15 Hz or four pulses at 200 Hz) . Carbon fiber electrodes were implanted in these regions to monitor evoked dopamine overflow by continuous amperometry. The kinetics of dopamine elimination were estimated by measuring the time to 50% decay of the dopamine oxidation current after stimulation ceased . This time ranged from 64 ms in the striatum to 113 ms in the nucleus accumbens. Inhibition of dopamine uptake by nomifensine (2-20 mg/kg), GBR 12909 (20 mg/kg), cocaine (20 mg/kg), mazindol (10 mg/kg), or bupropion (25 mg/kg) enhanced this decay time by up to +602% . Uptake inhibition also produced an increase in the maximal amplitude of dopamine overflow evoked by four pulses at 15 Hz . This latter effect was larger in the striatum (+420%) than in mesolimbic areas (+140%) . These results show in vivo that these uptake inhibitors actually slow the clearance of dopamine released by action potentials and suggest that dopaminergic transmission is both prolonged and potentiated strongly by these drugs, in particular in the striatum .

The paired phasic and tonic motor neurons supplying the extensor muscle in the crayfish leg were investigated to establish whether differences in synaptic structure could account for large differences in transmitter release at the neuromuscular junctions. Nerve terminals with transmitter release that had been assessed from recordings made with a focal "macro-patch" electrode were subsequently labeled, processed for electron microscopy, and reconstructed from serial sections. At a frequency of 1 Hz, quantal contents of phasic terminals were 90-1300 times greater than those of tonic terminals when both were recorded at the same location. At higher frequencies, facilitation was pronounced at tonic, but not phasic, terminals. Reconstructions of recording sites showed that both phasic and tonic terminals possessed many small synapses, usually with one or more structurally defined active zones. Mean synaptic contact area was larger for tonic terminals, and the number of individual synapses per length of nerve terminal was also larger. Active zones were not different in size for the two terminals. At low frequencies, quantal emission per synapse is much greater for phasic terminals. The higher quantal content of phasic terminals and their synapses cannot reasonably be accounted for by more or larger synapses or active zones at the recording sites. Because structural features alone are not likely to produce the very large differences in quantal content of phasic and tonic terminals observed at low stimulation frequencies, it is likely that other properties of the nerve terminal are largely responsible for these differences.

Synaptic functional differentiation of crayfish phasic and tonic motor neurons is large. For one impulse, quantal release of neurotransmitter is typically 100-1000 times higher for phasic synapses. We tested the hypothesis that differences in synaptic strength are determined by differences in synaptic calcium entry. Calcium signals were measured with the injected calcium indicator dyes Calcium Green-1 and fura-2. Estimated Ca(2+) entry increased almost linearly with frequency for both axons and was two to three times larger in phasic terminals. Tonic terminal Ca(2+) at 10 Hz exceeded phasic terminal Ca(2+) at 1 Hz, yet transmitter release was much higher for phasic terminals at these frequencies. Freeze-fracture images of synapses revealed on average similar numbers of prominent presynaptic active zone particles (putative ion channels) for both neurons and a two- to fourfold phasic/tonic ratio of active zones per terminal volume. This can account for the larger calcium signals seen in phasic terminals. Thus, differences in synaptic strength are less closely linked to differences in synaptic channel properties and calcium entry than to differences in calcium sensitivity of transmitter release.

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The aim of this study was to investigate whether or not nerve impulses release ATP and noradrenaline in parallel from the sympathetic nerve terminals of the rat tail artery. The extracellularly recorded excitatory junction current (EJC) was used to study, pulse by pulse, the release of ATP. An electrochemical method was used to study online the nerve stimulation-induced rise in the extracellular concentration of endogenous noradrenaline at the probe, a carbon fibre electrode (CF). This parameter, which does not directly represent noradrenaline release, but reflects release minus clearance, has been termed delta[NA]CF. The effects of a number of pharmacological agents on the EJCs were examined both at 0.1 and 2 Hz, and the effects on the EJC response to 100 pulses at 2 Hz compared with that on the delta[NA]CF response. Clonidine and xylazine were used as alpha 2-agonists, yohimbine and idazoxan as alpha 2-antagonists and desipramine and cocaine as blockers of noradrenaline reuptake. Most of these agents had unwanted side effects, especially at higher concentrations. However, clonidine and xylazine depressed at lower concentrations the EJC and delta[NA]CF responses to about the same extent; these effects were partially or completely reversed by yohimbine. Yohimbine or idazoxan did not affect the EJCs at 0.1 Hz but enhanced the EJC and delta[NA]CF responses to 100 pulses at 2 Hz to the same extent. All effects of desipramine (1 microM) seemed explainable as a result of block of noradrenaline reuptake, while cocaine (10 microM) in addition exerted an 'unspecific' depressant (probably local anesthetic) effect. Under control conditions, both agents depressed the EJC but dramatically enhanced the delta[NA]CF response to 100 pulses at 2 Hz. Addition of yohimbine prevented the depressant effect of desipramine on the EJCs completely and reduced that of cocaine, but increased their effects on the delta[NA]CF response. These results are compatible with the view that ATP and noradrenaline are released in parallel from the sympathetic nerve terminals of this tissue. The different, and under some conditions even opposite, effects of desipramine or cocaine on the EJC and delta[NA]CF responses are explainable in terms of the known post-secretory effects of these agents.