No abstract

The influence of melatonin on hippocampal evoked potentials initiated by low- and high-frequency electrical stimulations and by two pulses applied in rapid succession was investigated. In confirmation of our previous studies, melatonin attenuated the population spike triggered by low-frequency stimulation (0.03 Hz). High-frequency stimulation (HFS; 100 Hz for 1 sec, three times every 10 sec), which in control slices permanently facilitated neuronal excitability (347% +/- 32%), was also able to amplify the melatonin-depressed potential (467.8% +/- 59.6%). Because melatonin is a hydrophobic molecule, it was dissolved and applied in ethanol. Ethanol (0.4%) by itself reduced the magnitude of HFS-induced potentiation (233.5% +/- 16.8%). The slices stimulated with two pulses separated with a delay longer than 15 msec demonstrated a facilitation of the response to the second stimuli (paired-pulse facilitation; PPF). The influence of melatonin (100 microM) on PPF was biphasic: Shortly after addition of melatonin, PPF was briefly (5-10 min) reversed to paired-pulse inhibition (PPI), which gradually returned to a stable PPF. Ethanol (0.4%) applied without melatonin exerted only a marginal, facilitatory effect on PPF. The delay between two successively applied pulses, shorter than 13 msec, resulted in attenuation of the response to the second stimuli (PPI). Melatonin (100 microM) reversed the attenuation of the second potential within 15-20 min following its application. Ethanol applied by itself at the concentration of 0.4% temporarily (5-10 min), but significantly, depressed the second potential. These results demonstrate the ability of melatonin to modulate specific forms of plasticity in hippocampal pyramidal neurons.

The influence of melatonin on evoked potentials recorded from the CAI field of mouse hippocampal slices was investigated. Melatonin (0.1-2.0 mM) and its analog, 6-chloromelatonin (0.1-0.5 mM) depressed evoked potentials (EPSP and the population spike) in a concentration-dependent manner. The melatonin-induced depression was followed by a slow recovery phase. Since the fiber potential was not affected, it was concluded that melatonin influenced synaptic efficiency and/or cell excitability. Luzindole, an antagonist of MT2 melatonin receptors, although slightly depressing evoked potentials when applied by itself (100 microM), blocked any further inhibition by melatonin when added afterwards. We concluded that melatonin reduced synaptic efficiency and/or excitability of hippocampal neurons most likely through interaction with MT2 melatonin receptors, but other possible mechanisms of melatonin action are also considered.

During the induction of long-term potentiation (LTP) in hippocampal slices adenosine triphosphate (ATP) is secreted into the synaptic cleft, and a 48 kDa/50 kDa protein duplex becomes phosphorylated by extracellular ATP. All the criteria required as evidence that these two proteins serve as principal substrates of ecto-protein kinase activity on the surface of hippocampal pyramidal neurons have been fulfilled. This phosphorylation activity was detected on the surface of pyramidal neurons assayed after synaptogenesis, but not in immature neurons nor in glial cells. Addition to the extracellular medium of a monoclonal antibody termed mAb 1.9, directed to the catalytic domain of protein kinase C (PKC), inhibited selectively this surface protein phosphorylation activity and blocked the stabilization of LTP induced by high frequency stimulation (HFS) in hippocampal slices. This antibody did not interfere with routine synaptic transmission nor prevent the initial enhancement of synaptic responses observed during the 1-5 min period immediately after the application of HFS (the induction phase of LTP). However, the initial increase in the slope of excitatory postsynaptic potentials, as well as the elevated amplitude of the population spike induced by HFS, both declined gradually and returned to prestimulus values within 30-40 min after HFS was applied in the presence of mAb 1.9. A control antibody that binds to PKC but does not inhibit its activity had no effect on LTP. The selective inhibitory effects observed with mAb 1.9 provide the first direct evidence of a causal role for ecto-PK in the maintenance of stable LTP, an event implicated in the process of learning and the formation of memory in the brain.The long-term potentiation (LTP) of synaptic strength in hippocampal pyramidal neurons is a neurophysiological process iinplicated in the formation of memory traces in the brain (1, 2). While the initial chain of events that triggers this process is well-known, the mechanisms that determine the duration and stability of LTP have not yet been fully elucidated. Although it is generally accepted that the phosphorylation of proteins by several different kinases is involved in this stabilization, their exact localization and roles are still obscure (2, 3). Protein phosphorylation, a ubiquitous step in intracellular pathways that produce transient changes in neuronal activity, was found to serve also as a key mechanism of molecular adaptation in processes underlying the induction of longlasting alterations in synaptic function (for reviews, see ref. 4), including learning and memory formation (5-7). The first study that implicated protein phosphorylation in the process of learning (5) identified a synaptic phosphoprotein that was later shown to be a major neuronal substrate of protein kinase C (PKC), that plays a role in the maintenance-phase of LTP (8, 9). The specific PKC-isozyme involved in the maintenance of LTP was reported to be PKC C, a member of the atypical class (not stimulated by diacylglycerol or phorbol ...