Diurnal actions of melatonin on epileptic activity in hippocampal slices of rats

Since melatonin receptors have been found in the hippocampus of mammals it has been suggested that melatonin can modulate neuronal functions of hippocampal cells. The effect of melatonin (10 nM/l and 1 microM/l) on frequency and amplitude of epileptiform field potentials (EFP) elicited by low Mg(2+) or by bicuculline was tested in the CA1 region of hippocampal slices of rats. In the low Mg(2+) model, melatonin, applied in a near physiological concentration of 10 nM/l, exerts no effect on EFP in slices prepared at night or during the day. In a concentration of 1 microM/l, however, melatonin enhances the frequency of EFP to approximately 140% in slices prepared during the day. This effect was suppressed through simultaneous administration of the melatonin receptor antagonist luzindole (10 microM/l). In contrast, melatonin did not affect epileptic activity in slices prepared at night. Epileptiform discharges elicited by blocking the GABAergic inhibition (bicuculline model) were not affected by melatonin, either during the day or at night. The results indicate that melatonin affects epileptic activity in a diurnal manner and that the action of melatonin is different in relation to the epilepsy model.     

Effects of adrenal steroids and their reduced metabolites on hippocampal long-term potentiation

We studied the effects of steroid hormones on the hippocampal long-term potentiation (LTP), a putative mechanism of neuronal plasticity and memory storage in the CNS. In vivo experiments were performed in rats under chloral hydrate anesthesia (0.4 mg/kg i.p.). All animals were adrenalectomized 48 h before recording. LTP was induced after priming tetanic stimulation at the perforant pathway (PP) and single pulse field potentials were obtained from the dentate gyrus (DG). The excitatory post-synaptic potential (EPSP) slope and population spike (PS) amplitude were analyzed before and after the i.v. injection of the steroids and after the induction of LTP, and followed up to 1 h. Results obtained with the hormones were compared with matched control animals injected with vehicle alone, Nutralipid 10%. Previous results from our laboratory showed that deoxycorticosterone (DOC) decreased the magnitude of the EPSP at all times after priming stimulation and the PS decreased during the first 30 min of the LTP. Corticosterone decreased the EPSP in the first 15 min and the PS during the first 30 min after priming stimuli. In these experiments the mineralocorticoids aldosterone and 18-OH-DOC elicited a decrease of the EPSP at all times post-train; and no significant difference against vehicle was observed in the PS. Post-injection values were not changed except for 18-OH-DOC at a dose of 1 mg, where a decrease of both the EPSP (P less than 0.01) and the PS (P less than 0.02) was observed against vehicle. ATH-progesterone at 0.1 mg/rat also decreased the EPSP values significantly after priming stimulation and no significant changes against vehicle were observed in the PS. These results show that adrenal steroids can modulate hippocampal LTP, that they can act at different neuronal loci and with different time courses in the development of the phenomena.     

Dantrolene modulates the influence of steady magnetic fields on hippocampal evoked potentials in vitro

Direct current-generated magnetic fields (2-3 mT, 20-min exposure) exerted biphasic effects on the population spike recorded from hippocampal slices. The initial decrease in the potential, observed during exposure of the slices to magnetic fields was followed by a recovery/amplification phase, which began after terminating the magnetic field action. During that phase the population spike exceeded the amplitude observed before application of the magnetic fields. The pattern of magnetic fields influence was not affected either by (+)-5-methyl-10,11-dihydro-5H-dibenzo (a,d) cyclohepten-5, 10-imine maleate (MK801), or by D,L,-2amino-5phosphonovalerate (APV), a noncompetitive and competitive NMDA receptor antagonist, respectively. The rising phase of the potential, however, was eliminated by dantrolene, an inhibitor of intracellular Ca(2 +) channels. This suggests that intracellular calcium channels participate in the mechanism of the influence of the direct current magnetic fields on the function of the hippocampal tissue.     

Effects of A1 and A2 Adenosine Receptor Antagonists on the Induction and Reversal of Long-Term Potentiation in Guinea Pig Hippocampal Slices of CA1 Neurons

1. Using simultaneous recordings of the field EPSP and the population spike in the CA1 neurons of guinea pig hippocampal slices, we confirmed that delivery of a high-frequency stimulation (tetanus: 100 pulses at 100 Hz) produced robust long-term potentiation of synaptic efficacy (LTP) in two independent components, a synaptic component that increases field excitatory postsynaptic potentials (EPSPs) and a component that results in a larger population spike amplitude for a given EPSP size (E-S potentiation).2. In the same cells, reversal of LTP (depotentiation; DP) in the field EPSP and in the E-S component is achieved by delivering low-frequency afferent stimulation (LFS:1 Hz, 1000 pulses) 20 min after the tetanus.3. When the tetanus or LFS was applied to CA1 inputs in the presence of an adenosine A₁ receptor antagonist, 8-cyclopentyltheophylline (1 M), the field EPSP was enhances in LTP and attenuated in DP, while the E-S relationship was not significantly affected in either LTP or DP.4. When similar experiments were performed using an A₂ receptor antagonist, CP-66713 (10 M), the field EPSP was blocked in LTP but facilitated in DP, while E-S potentiation was enhanced during both LTP and DP.5. The results show that endogenous adenosine, acting via A₁ or A₂ receptors, modulates both the synaptic and the E-S components of the induction and reversal of LTP. Based on the results, we discuss the key issue of the contribution of these receptors to the dynamics of neuronal plasticity modification in hippocampal CA1 neurons.     

Modulation by adrenal steroids of limbic function

The effects of various steroid hormones on the long-term potentiation (LTP) of the rat hippocampus were evaluated. LTP was elicited in the dentate gyrus of adrenalectomized animals with priming tetanic stimulation (200 Hz-0.03 cps) of its main afferent, the perforant pathway. Single pulse EPSP (excitatory post-synaptic potential) slope, and PS (population spike) amplitude values were compared before and after the i.v. injection of the hormones and subsequently after the priming stimulation every 15 min up to 1 h. 18-OH-deoxycorticosterone (18-OH-DOC) produced a significant decrease of the EPSP LTP and arrested the PS enhancement in comparison with vehicle at every time post-tetanic stimulation. Its 21-acetate derivative produced a moderate decrease of the EPSP and had no effect on the PS LTP in comparison with vehicle. Deoxycorticosterone (DOC) exhibited similar effects on the EPSP although less marked than with 18-OH-DOC while the PS only decreased in the first 30 min post-train. Corticosterone decreased both EPSP and PS for the first 15 and 30 min after priming stimulation, respectively, matching values with those of vehicle afterwards. Its 21-acetate produced an initial decrease of the EPSP and had no effect on the PS LTP. Allo-tetrahydro-DOC produced little, if any, initial enhancement of the PS LTP in comparison with vehicle. These results show that the adrenal steroids tested can modulate hippocampal LTP, a plastic phenomenon in the mammalian CNS which is known to be related to memory and learning processes. Moreover, adrenal steroids can independently modify the PS or EPSP components of the LTP, suggesting different loci of action at the neuronal level.     

Barium, glibenclamide and CGS21680 prevent adenosine A1 receptor changes of ES coupling and spike threshold

Activation of adenosine A1 receptors raised spike thresholds and induced a dissociation of excitatory postsynaptic potential (EPSP) spike coupling in hippocampal pyramidal neurones. This effect could be prevented by activation of A2A adenosine receptors. The A1 receptor agonist N6-cyclopentyladenosine caused a dissociation of the EPSP spike coupling recorded extracellularly and increased the threshold for spike generation measured intracellularly. These effects were prevented by the A2A receptor agonist CGS21680. A series of agents interfering with adenylate cyclase activity, protein kinase A or C, or nitric oxide synthase had no effect on these responses to N6-cyclopentyladenosine. Superfusion with barium or glibenclamide prevented both the dissociation of EPSP spike coupling and the increase of spike threshold. It is concluded that a barium- and glibenclamide-sensitive potassium current may be involved in the postsynaptic effects of A1 receptors on spike threshold, and it is suggested that a similar suppression of a potassium current by A2A receptors could underlie the inhibition of A1 receptor responses.     

A biphasic excitability change in hindlimb motoneurons evoked by stimulation of the nucleus raphes magnus in the cat

1. The influence of electrical stimulation of the nucleus raphes magnus (RM) on spinal segmental systems were examined. 2. RM stimulation produced an initial increase and a subsequent suppression of the amplitude of both fiextor and extensor lumbar monosynaptic reflex potentials (MSRs). 3. Intracellular recordings were made from alpha-motoneurons of the common peroneal nerve (flexor) and the tibial nerve (extensor). RM stimulation evoked postsynaptic potentials with a time course similar to that of MSR facilitation. 4. RM stimulation inhibited the aggregate excitatory synaptic potential (EPSP) evoked by stimulation of group I afferent fibers without apparent changes in the motoneuronal membrane potential. 5. These data suggest that the RM-evoked biphasic effect on MSR consists of early facilitation due to EPSP, and late inhibition possibly due to presynaptic inhibition of group I afferent fibers.     

Pregnenolone sulfate induces NMDA receptor dependent release of dopamine from synaptic terminals in the striatum

Neuromodulators that alter the balance between lower-frequency glutamate-mediated excitatory and higher-frequency GABA-mediated inhibitory synaptic transmission are likely to participate in core mechanisms for CNS function and may contribute to the pathophysiology of neurological disorders such as schizophrenia and Alzheimer's disease. Pregnenolone sulfate (PS) modulates both ionotropic glutamate and GABA(A) receptor mediated synaptic transmission. The enzymes necessary for PS synthesis and degradation are found in brain tissue of several species including human and rat, and up to 5 nM PS has been detected in extracts of postmortem human brain. Here, we ask whether PS could modulate transmitter release from nerve terminals located in the striatum. Superfusion of a preparation of striatal nerve terminals comprised of mixed synaptosomes and synaptoneurosomes with brief-duration (2 min) pulses of 25 nM PS demonstrates that PS increases the release of newly accumulated [3H]dopamine ([3H]DA), but not [14C]glutamate or [3H]GABA, whereas pregnenolone is without effect. PS does not affect dopamine transporter (DAT) mediated uptake of [3H]DA, demonstrating that it specifically affects the transmitter release mechanism. The PS-induced [3H]DA release occurs via an NMDA receptor (NMDAR) dependent mechanism as it is blocked by D-2-amino-5-phosphonovaleric acid. PS modulates DA release with very high potency, significantly increasing [3H]DA release at PS concentrations as low as 25 pM. This first report of a selective direct enhancement of synaptosomal dopamine release by PS at picomolar concentrations via an NMDAR dependent mechanism raises the possibility that dopaminergic axon terminals may be a site of action for this neurosteroid.     

EPSP amplification and the precision of spike timing in hippocampal neurons

The temporal precision with which EPSPs initiate action potentials in postsynaptic cells determines how activity spreads in neuronal networks. We found that small EPSPs evoked from just subthreshold potentials initiated firing with short latencies in most CA1 hippocampal inhibitory cells, while action potential timing in pyramidal cells was more variable due to plateau potentials that amplified and prolonged EPSPs. Action potential timing apparently depends on the balance of subthreshold intrinsic currents. In interneurons, outward currents dominate responses to somatically injected EPSP waveforms, while inward currents are larger than outward currents close to threshold in pyramidal cells. Suppressing outward potassium currents increases the variability in latency of synaptically induced firing in interneurons. These differences in precision of EPSP-spike coupling in inhibitory and pyramidal cells will enhance inhibitory control of the spread of excitation in the hippocampus.     

The influence of pulsed magnetic fields (PMFs) on nonsynaptic potentials recorded from the central and peripheral nervous systems in vitro

The influence of pulsed magnetic fields (PMFs) on nonsynaptic potentials recorded from the central and peripheral nervous system in vitro has been investigated. The population spikes (PSs) recorded from hippocampal slices during antidromic stimulation and compound action potentials (CAPs) recorded from the segments of the sciatic nerve were used as indicators of neuronal activity. The potentials recorded from both preparations were significantly and permanently enhanced following PMF (0.16 Hz, 15 mT) exposure. The increase in the antidromic PS occurred even in the presence of potassium channel blocker tetraethylammonium (TEA) and was accompanied by multiple spiking. Among all frequencies of PMF tested (0.5, 0.16, 0.07, 0.03, 0.0 Hz), the frequency of 0.5 Hz was the most effective in enhancement of potential amplitude. The influence of PMF on the amplitude of two CAPs evoked by the pair of electrical stimuli applied in rapid succession has also been evaluated. In control conditions the potential triggered by the second stimuli was slightly smaller expressing the phenomenon of short‐term depression (STD). Although PMF exposure amplified the amplitude of both potentials, the increase in the size of the first potential was significantly greater increasing further the magnitude of STD. The blocking of potassium channels reversed STD into facilitation. One of the possible mechanisms involved in PMF action could be the modification of the axonal threshold, which was significantly reduced following exposure to PMF. Bioelectromagnetics 30:621–630, 2009. © 2009 Wiley‐Liss, Inc.     

The neurosteroids pregnenolone and pregnenolone-sulfate but not progesterone, block Ca2+ currents in acutely isolated hippocampal CA1 neurons.

The neurosteroids pregnenolone (PE) and pregnenolone-sulfate (PS) have been shown to interact with the GABAA receptor in the central nervous system. In contrast, nothing is known of any possible modulation of voltage-gated calcium channels (VGCC). We have examined the interaction of PE, PS and progesterone on VGCC in acutely isolated adult guinea-pig hippocampal CA1 neurons using the whole-cell patch clamp technique. PE and PS depressed the calcium current at low micromolar concentrations (0.001-100 microM). The time to peak of the calcium current was slowed by PE and PS. The blocking action of PE and PS occurs in the presence of 10 microM picrotoxin. In contrast, progesterone had no effect on the Ca2+ current, indicating specificity for PE and PS. These results demonstrate a direct and novel membrane site of action for PE and PS, suggesting a possible role influencing brain excitability.     

Clozapine functions through the prefrontal cortex serotonin 1A receptor to heighten neuronal activity via calmodulin kinase II-NMDA receptor interactions

Aberrant dopamine release in the prefrontal cortex (PFC) is believed to underlie schizophrenia, but the mechanistic pathway through which a widely used antipsychotic, clozapine (Clz), evokes neurotransmitter-releasing electrical stimulation is unclear. We analyzed Clz-evoked regulation of neuronal activity in the PFC by stimulating axons in layers IV and V and recording the electrical effect in the post-synaptic pyramidal cells of layers II and III. We observed a Clz-evoked increase in population spike (PS), which was mediated by serotonin 1A receptor (5-HT(1A)-R), phospholipase Cβ, and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). Immunoblotting demonstrated that the Clz-activation of CaMKII was 5-HT(1A)-R-mediated. Intriguingly, the NMDA receptor (NMDA-R) antagonist (±)2-amino-5-phosphonovaleric acid (APV) eliminated the Clz-mediated increase in PS, suggesting that the 5-HT(1A)-R, NMDA-R and CaMKII form a synergistic triad, which boosts excitatory post-synaptic potential (EPSP), thereby enhancing PS. In corroboration, Clz as well as NMDA augmented field EPSP (fEPSP), and WAY100635 (a 5-HT(1A)-R antagonist), APV, and a CaMKII inhibitor eliminated this increase. As previously shown, CaMKII binds to the NMDA-R 2B (NR2B) subunit to become constitutively active, thereby inducing α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) receptor recruitment to the post-synaptic membrane and an increase in fEPSP. Co-immunoprecipitation demonstrated that Clz potentiates interactions among CaMKII, NR2B, and 5-HT(1A)-R, possibly in the membrane rafts of the post-synaptic density (PSD), because pretreatment with methyl-β-cyclodextrin (MCD), an agent that disrupts rafts, inhibited both co-immunoprecipitation as well as fEPSP. In summary, Clz functions in the PFC by orchestrating a synergism among 5-HT(1A)-R, CaMKII, and NMDA-R, which augments excitability in the PFC neurons of layers II/III.     

Dexamethasone modulates melatonin MT2 receptor expression in splenic tissue and humoral immune response in mice

Melatonin modulates immune function through its membrane-bound MT1 and MT2 receptors in mammalian system. Adrenal glucocorticoid, an important metabolic hormone is known as a immuno-compromising agent. In the present study, we investigated the effect of dexamethasone on melatonin receptor proteins in spleen tissue and anti-klh-IgG response in Swiss albino mice. Melatonin treatment increased the MT1 and MT2 receptor proteins and anti-klh-IgG than control mice. Dexamethasone treatment increased MT2 receptor protein and anti-klh-IgG than melatonin-treated group. Dexamethasone treatment to melatonin-treated mice showed additive effects and maximally increased the anti-klh-IgG than other experimental groups. A decrease in glucocorticoid receptor (GR) protein was noted in melatonin treated as well as dexamethasone-treated mice. Dexamethasone significantly increased MT2 melatonin receptor protein in spleen and anti-klh-IgG and additively increased anti-klh-IgG when supplemented along with melatonin. Therefore, the present study may suggest that dexamethasone increased humoral immune response permissively by enhancing MT2 receptor expression in splenic tissue of mice.     

Circadian system functionality, hippocampal oxidative stress, and spatial memory in the APPswe/PS1dE9 transgenic model of Alzheimer disease: effects of melatonin or ramelteon

Alzheimer disease (AD) is a neurodegenerative disorder that primarily causes β-amyloid accumulation in the brain, resulting in cognitive and behavioral deficits. AD patients, however, also suffer from severe circadian rhythm disruptions, and the underlying causes are still not fully known. Patients with AD show reduced systemic melatonin levels. This may contribute to their symptoms, since melatonin is an effective chronobiotic and antioxidant with neuroprotective properties. Here, the authors critically assessed the effects of long-term melatonin treatment on circadian system function, hippocampal oxidative stress, and spatial memory performance in the APPswe/PS1 double transgenic (Tg) mouse model of AD. To test if melatonin MT1/MT2 receptor activation, alone, was involved, the authors chronically treated some mice with the selective MT1/MT2 receptor agonist ramelteon. The results indicate that many of the circadian and behavioral parameters measured, including oxidative stress markers, were not significantly affected in these AD mice. During the day, though, Tg controls (Tg-CON) showed significantly higher mean activity and body temperature (BT) than wild-type (WT) mice. Overall, BT rhythm amplitude was significantly lower in Tg than in WT mice. Although melatonin treatment had no effect, ramelteon significantly reduced the amplitude of the BT rhythm in Tg mice. Towards the end of the experiment, Tg mice treated with ramelteon (Tg-RAM) showed significantly higher circadian rhythm fragmentation than Tg-CON and reduced circadian BT rhythm strength. The free-running period (τ) for the BT and locomotor activity (LA) rhythms of Tg-CON was <24 h. Whereas melatonin maintained τ at 24 h for BT and LA in both genotypes, ramelteon treatment had no effect. In the behavioral tests, the number of approaches and time spent exploring novel objects were significantly higher in Tg-CON than WT controls. Brain tissue analysis revealed significant reduction in hippocampal protein oxidation in Tg-MEL and Tg-RAM compared with Tg-CON animals. These results suggest that not all aspects of the circadian system are affected in the APPswe/PS1 mice. Therefore, care should be taken when extending the results obtained in Tg mice to develop new therapies in humans. This study also revealed the complexity in the therapeutic actions of melatonin and ramelteon in this mouse model of AD.     

在体大鼠海马CA1区侧枝/联合纤维通路和TA通路的不同EPSP空间整合特性

在麻醉Wistar大鼠上,结合脑室给药,应用双电极刺激技术刺激海马独立的两条侧枝／联合纤维通路、TA通路,并在CA1区放射层记录兴奋性突触后电位（EPSP）,对海马CA1区锥体细胞近、远端树突EPSP的空间整合进行了初步探讨。结果表明,海马CA1区锥体细胞近、远端树突的空间整合都是亚线性的;近端树突的空间整合不受期望值大小的影响,但远端树突的空间整合随期望值增加而减小（更趋于亚线性）。此外,荷包牡丹碱没有影响EPSP的空间整合;但瞬时A型钾通道（IAK^＋）的拮抗剂氨基吡啶-4却使得近端树突的空间整合趋于线性发展。本研究表明,海马CA1锥体细胞近、远端树突不同的被动、主动特征使它们具有了不同的空间整合特性。由于近端树突接受海马内部侧枝／联合纤维投射的信息,远端树突通过TA通路接受内嗅皮层投射的信息,由此提示,CA1区锥体细胞对来自海马内部和直接来自皮层的信息输入采用了不同的整合方式。     

Adenosine (A2) antagonist inhibits induction of long-term potentiation of evoked synaptic potentials but not of the population spike in hippocampal CA1 neurons.

The effects of adenosine A2 receptor antagonist (CP-66713) on long-term potentiation were studied using guinea pig hippocampal slices in a perfusion system. Tetanic stimulation of Schaffer collateral input which was applied during perfusion of CP-66713 (10 microM), did not induce long-term potentiation but rather long-term depression of evoked synaptic potentials (field EPSP), but induced long-term potentiation of the population spike in CA1 neurons. Thus, adenosine derivatives which accumulate in the synaptic cleft during the tetanic stimulation may be involved in induction of the long-term potentiation via A2 receptors at the synapse. The clear discrimination between long-term depression of the field EPSP and long-term potentiation of the population spike suggests EPSP-spike potentiation at the postsynaptic sites.     

Neuromodulatory effect of creatine on extracellular action potentials in rat hippocampus: role of NMDA receptors

The creatine (Cr) and phosphocreatine (PCr) system is essential for the buffering and transport of high-energy phosphates. Although achievements made over the last years have highlighted the important role of creatine in several neurological diseases, the adaptive processes elicited by this guanidino compound in hippocampus are poorly understood. In the present study, we showed that creatine (0.5-25mM) gradually increases the amplitude of first population spike (PS) and elicits secondary PS in stratum radiatum of the CA1 region, in hippocampal slices. Creatine also decreased the intensity of the stimulus to induce PS, when compared with hippocampal slices perfused with artificial cerebrospinal fluid (ACSF). The competitive NMDA receptor antagonist, 2-amino-5-phosphonopentanoic acid (AP5; 100microM) attenuated creatine-induced increase of amplitude of PS and appearance of secondary PS, providing pharmacological evidence of the involvement of NMDA receptors in the electrophysiological effects of creatine. Accordingly, creatine (0.01-1mM) increased [3H]MK-801 binding to hippocampal membranes by 55%, further indicating that this compound modulates NMDA receptor function. These results implicate the NMDA receptor in amplitude and population spike increase elicited by creatine in hippocampus. Furthermore, these data suggest that this guanidino compound may also play a putative role as a neuromodulator in the brain, and that at least some of its effects may be mediated by an increase in glutamatergic function.     

Influence of oxytocin on integration of postsynaptic potentials in molluscan neurons.

1. Intracellular recordings were made from identified and non-identified neurons in perioesophageal ganglionic ring with buccal ganglia of the mollusc, Helix pomatia. The influence of oxytocin (OXT) on neuronal integration: space and temporal summations of postsynaptic potentials (PSPs) in various neurons was investigated. The obtained data indicated that these PSPs were cholinergic PSPs. 2. Ten minute exposure to 10(-8) M OXT had no effects on the resting membranes, but triggered secondary mechanisms, which lead to enhancement of the excitatory PSP (EPSR) amplitudes and the decrease of the decay time constant (tau EPSR) obtained from the falling phase of the EPSP. 3. The enhancement of the EPSP amplitude and the decrease of tau EPSP after OXT application evoked the appearance of action potential under space summation of two spontaneous EPSPs and made easier the appearance of action potential under temporal summation of EPSPs produced by paired afferent stimuli, when the corresponding interstimuli interval was smaller than tau EPSP in the presence of OXT. 4. Ten minute exposure to 10(-8) M OXT made the integrated amplitude of the excitatory acetylcholine response and the inhibitory dopamine response in the neuron E5 more positive only when the interval between applications of these mediators was smaller than the time constant of desensitization of acetylcholine receptors in the presence of OXT. 5. The pharmacological studies showed that drugs, which elevate intracellular cyclic AMP levels, mimicked the influence of OXT on integration of PSPs in the investigated neurons.     

Effects of extracellular calcium on low frequency induced potentiation and habituation in the in vitro hippocampal slice preparation

The electrophysiological characteristics of frequency potentiation and habituation were investigated in two afferent systems of the in vitro hippocampal slice preparation. Low frequency stimulation (1 Hz) of the Schaffer collateral - commissural (Sch-comm) fibers results in a short-term potentiation of the amplitude and rate of rise of the EPSP and population spike responses recorded in the CA1 region. In contrast, 1-Hz stimulation of the perforant path (PP) evokes a short-term, habituation-like depression of the dentate granule cell EPSP and population spike. An inverse relationship was observed between stimulus intensity and the magnitude of frequency potentiation or habituation. Changes in afferent fiber volleys or general excitability of postsynaptic membranes did not contribute significantly to the development of either of these forms of short-term plasticity. Perfusion with a medium containing a high calcium - low magnesium concentration (4 mM Ca+2 and 1 mM Mg+2) produced a differential effect on CA1 and dentate evoked potentials. Following a 20-min exposure to this medium, the amplitude of CA1 potentials was increased while dentate responses were decreased. Frequency potentiation of CA1 responses and habituation of dentate responses were depressed or eliminated by the high calcium medium. The opposing influence of extracellular calcium on CA1 and dentate evoked potentials indicates a fundamental difference in the process of transmitter release in these systems, a characteristic that may contribute to the production of frequency potentiation and habituation.