Area CA1 Of Hippocampal Slices Research Articles

Differential effects of cation-chloride co-transport-blocking diuretics in a rat hippocampal slice model of epilepsy

The cation-Cl − co-transport-blocking loop diuretics have clinically known anticonvulsant activity, though they can also induce seizures. We explored the effects of ethacrynic acid (ETA), furosemide (FUR) and bumetanide (BUM), prototypical blockers of cation-Cl − co-transport, on the epileptiform field potentials induced in CA3 area of hippocampal slices from 5-weeks-old rats by a high K +–low Ca 2+ perfusion fluid. That milieu induces frequent spontaneous field bursts, making single fimbrial stimuli to evoke several repetitive population spikes, of increased amplitude. ETA (0.25–1 mM) concentration-dependently reduced spontaneous field bursting, up to terminating it. FUR, 5 mM also inhibited spontaneous field bursting, while BUM (12.5–100 μM) only presented an inconsistent tendency. Both ETA and FUR showed a less marked ability to depress the evoked responses, but ∼mM concentrations significantly reduced the number of repetitive population spikes and their amplitude. BUM only modestly reduced population spike amplitude, without concentration-dependence. This study shows that K +–Cl − co-transport-blocking diuretics ETA and FUR inhibit high K +-induced epileptiform activity in hippocampal slices from (nearly) adult rats, while the Na +–K +–2Cl − co-transport-preferring diuretic BUM had only negligible activity. These results support that neuronal K +–Cl − co-transport-blockade provides antiepileptic effects.

Chronic hyperammonemia in vivo impairs long‐term potentiation in hippocampus by altering activation of cyclic GMP‐dependent‐protein kinase and of phosphodiesterase 5

Long-term potentiation (LTP) is impaired in the CA1 area of hippocampal slices from rats with chronic moderate hyperammonemia. We studied the mechanisms by which hyperammonemia in vivo impairs LTP. This process requires sequential activation of soluble guanylate cyclase, cyclic GMP-dependent protein kinase (PKG) and cyclic GMP-degrading phosphodiesterase. Application of the tetanus induced a rapid increase of cyclic GMP in slices from control or hyperammonemic rats, which is followed in control slices by a sustained decrease in cyclic GMP due to sustained activation of cyclic GMP-degrading phosphodiesterase, which in turn is due to sustained activation of PKG. In slices from rats with chronic hyperammonemia tetanus-induced decrease in cyclic GMP was delayed and transient due to lower and transient activation of PKG and of the phosphodiesterase. Hyperammonemia-induced impairment of LTP may be involved in the alterations of cognitive function in patients with hepatic encephalopathy.

Adenylate Cyclase-Mediated Forms of Neuronal Plasticity in Hippocampal Area CA1 Are Reduced With Aging

Beta-adrenergic receptors and the cyclic AMP signaling pathway play an important role in neuronal plasticity and in learning and memory and are known to change with aging. We examined the effects of beta-adrenergic stimulation paired with 5-Hz low frequency stimulation (LFS) of Schaffer collateral-commissural afferents on population spike amplitude in area CA1 of hippocampal slices from young (3 mo) and aged (22 mo) Fischer 344 rats. Application of the beta-adrenergic agonist isoproterenol (1 microM) for 10 min followed immediately by 3 min LFS produced long-lasting potentiation in young hippocampi, but the magnitude of potentiation in aged rats was significantly attenuated and was not long-lasting. In slices prepared from young rats, long-term potentiation (LTP) induced by this protocol occludes subsequent attempts to produce conventional high frequency stimulation-induced LTP, and vice versa, suggesting that these two forms of potentiation share one or more molecular mechanisms. Age-related differences in response to LFS alone were not observed, but significant differences in response to beta-adrenergic stimulation were apparent. Similarly, significant age-related differences in response to direct activation of adenylate cyclase with forskolin (10 microM) were observed. In both age groups, this enhancement produced by isoproterenol or forskolin is only transient, returning to baseline within 60 or 90 min, respectively. Taken together, these studies of adenylate cyclase-mediated forms of potentiation in area CA1 suggest that there is an age-related defect, either upstream or downstream of adenylate cyclase activation, in this important signaling system. Such changes may contribute to the compromised performance on memory tasks that is often observed with normal aging.

Adenylate Cyclase-Mediated Forms of Neuronal Plasticity in Hippocampal Area CA1 Are Reduced With Aging

Beta-adrenergic receptors and the cyclic AMP signaling pathway play an important role in neuronal plasticity and in learning and memory and are known to change with aging. We examined the effects of β-adrenergic stimulation paired with 5-Hz low frequency stimulation (LFS) of Schaffer collateral-commissural afferents on population spike amplitude in area CA1 of hippocampal slices from young (3 mo) and aged (22 mo) Fischer 344 rats. Application of the β-adrenergic agonist isoproterenol (1 μM) for 10 min followed immediately by 3 min LFS produced long-lasting potentiation in young hippocampi, but the magnitude of potentiation in aged rats was significantly attenuated and was not long-lasting. In slices prepared from young rats, long-term potentiation (LTP) induced by this protocol occludes subsequent attempts to produce conventional high frequency stimulation-induced LTP, and vice versa, suggesting that these two forms of potentiation share one or more molecular mechanisms. Age-related differences in response to LFS alone were not observed, but significant differences in response to β-adrenergic stimulation were apparent. Similarly, significant age-related differences in response to direct activation of adenylate cyclase with forskolin (10 μM) were observed. In both age groups, this enhancement produced by isoproterenol or forskolin is only transient, returning to baseline within 60 or 90 min, respectively. Taken together, these studies of adenylate cyclase-mediated forms of potentiation in area CA1 suggest that there is an age-related defect, either upstream or downstream of adenylate cyclase activation, in this important signaling system. Such changes may contribute to the compromised performance on memory tasks that is often observed with normal aging.

Tonic release of glutamate by a DIDS‐sensitive mechanism in rat hippocampal slices

Tonic release of glutamate into the extracellular space of the hippocampus and striatum is non-vesicular, and has been attributed largely to a cystine-glutamate exchanger which is blockable by the glutamate analogue (S)-4-carboxyphenylglycine (CPG). Tonic glutamate release may be functionally important: modulation of this release in the striatum has been suggested to underlie relapse in the use of cocaine. We monitored tonic glutamate release in area CA1 of hippocampal slices by measuring the glutamate receptor-mediated current evoked in pyramidal cells on block of Na(+)-dependent glutamate uptake with dl-threo-beta-benzyloxyaspartate (TBOA). Superfused cystine increased tonic glutamate release, and this increase was blocked by CPG, but CPG did not affect tonic glutamate release in the absence of superfused cystine. Tonic glutamate release was not affected by blocking gap junctional hemichannels with 18alpha-glycyrrhetinic acid, blocking ATP receptors with pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), blocking Ca(2)(+)-dependent exocytosis from neurones with Cd(2)(+) or bafilomycin, blocking Ca(2)(+)-dependent release from glia with indomethacin, or blocking anion channels with 5-nitro-2-(3-phenylpropyl amino) benzoic acid (NPPB) or tamoxifen. However tonic glutamate release was reduced by 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS), and was potentiated by inhibiting astrocytic conversion of glutamate to glutamine with methionine sulfoximine. These data suggest that although cystine-glutamate exchange is present in the hippocampus it does not generate significant tonic release of glutamate when the extracellular [cystine] is at a physiological level, and that tonic glutamate release is at least partly from astrocytes and is mediated by a DIDS-sensitive mechanism. Theoretical calculations suggest that a significant fraction of tonic glutamate release in hippocampal slices could occur via diffusion of glutamate across lipid membranes.

A pentylenetetrazole‐induced generalized seizure in early life enhances the efficacy of muscarinic receptor coupling to G‐protein in hippocampus and neocortex of adult rat

We have previously shown that exposure to the anti-cholinesterase eserine provokes interictal-like discharges in the CA3 area of hippocampal slices from adult rats in which a generalized seizure has been induced by pentylenetetrazole (PTZ) when immature (at 20 days). Such increased responsiveness to acetylcholine (ACh) was not associated with any change in hippocampal acetylcholine or gamma-aminobutyric acid (GABA) content, GABAergic inhibition or density of ACh innervation, but was blocked by the muscarinic receptor antagonist atropine. We therefore turned to quantitative radioligand binding autoradiography, in situ hybridization and the [35S]GTPgammaS method to assess the properties of hippocampal and neocortical muscarinic receptors in adult rats having experienced a PTZ seizure at P20. The densities of M1 and M2 receptor binding sites, respectively labeled with [3H]pirenzepine and [3H]AFDX-384, as well as the amount of m1, m2 and m3 receptor mRNAs, did not differ from control in the hippocampus and neocortex of these rats. In contrast, in PTZ rats, both brain regions displayed a marked increase in [35S]GTPgammaS incorporation stimulated by ACh, bethanechol and particularly oxotremorine. This finding indicates that a generalized seizure in immature rat can entail a long-term and presumably permanent increase in the efficacy of G-protein coupling to muscarinic receptors in the hippocampus and neocortex of the adult. By analogy, such a mechanism could account for the susceptibility to epilepsy of human adults having suffered from prolonged convulsions in early life.

Impairment in Short-Term but Enhanced Long-Term Synaptic Potentiation and ERK Activation in Adult Hippocampal Area CA1 Following Developmental Thyroid Hormone Insufficiency*

Thyroid hormones are critical for the development and maturation of the central nervous system. Insufficiency of thyroid hormones during development impairs performance on tasks of learning and memory that rely upon the hippocampus and impairs synaptic function in young hypothyroid animals. The present study was designed to determine if perturbations in synaptic function persist in adult euthyroid animals exposed developmentally to insufficient levels of hormone. Pre- and postnatal thyroid hormone insufficiency was induced by administration of 3 or 10 ppm propylthiouracil (PTU) to pregnant and lactating dams via the drinking water from gestation day (GD) 6 until postnatal day (PN) 30. This regimen produced a graded level of hormonal insufficiency in the dam and the offspring. Population spike and population excitatory postsynaptic potentials (EPSP) were recorded at the pyramidal cell layer and the stratum radiatum, respectively, in area CA1 of hippocampal slices from adult male offspring. PTU exposure increased baseline synaptic transmission, reduced paired-pulse facilitation, and increased the magnitude of the population spike long-term potentiation (LTP). Phosphorylation of the extracellular signal-regulated kinases (ERK1 and ERK2) was increased as a function of LTP stimulation in slices from PTU-exposed adult animals. On the other hand, no differences in the basal levels of synaptic proteins implicated in synaptic plasticity (total ERK, synapsin, growth-associated protein-43, and neurogranin) were detected. These results reinforce previous findings of persistent changes in synaptic function and, importantly extend these observations to moderate levels of thyroid hormone insufficiency that do not induce significant toxicity to the dams or the offspring. Such alterations in hippocampal synaptic function may contribute to persistent behavioral deficits associated with developmental hypothyroidism.

D-Serine enhances impaired long-term potentiation in CA1 subfield of hippocampal slices from aged senescence-accelerated mouse prone/8

The molecular and cellular mechanisms underlying the cognitive deficiency of senescence-accelerated mouse prone/8 (SAMP8) have been attributed to many pathological changes in neurons. Recently, increasing evidence has shown that astrocytes, by mean of d-serine, involve in the process of synaptic transmission. Here we reported that the long-term potentiation (LTP) in CA1 area of hippocampal slices prepared from 2-, 6- and 12-month-old SAMP8 significantly decreased with age. Meanwhile, the LTP in the slices of 6- and 12-month-old mice markedly decreased below that of the age-matched normal strain SAMR1. Supplement with exogenous d-serine, a main product of astrocytes and a coagonist at the “glycin-binding” site of N-methyl- d-aspartate (NMDA) receptors, not only directly enhanced the deficient LTP but also rescued the abolished LTP by d-amino acid oxidase (DAAO) in slices from 12-month-old SAMP8. This ameliorative effect of d-serine was inhibited by either AP-V or 5,7-dichlorokynurenic acid (DCKA). These results suggest that absence of d-serine or dysfunction of the astrocytes possibly was one of mechanisms underlying the decrease of NMDA receptor-dependent LTP and cognition in aged SAMP8.

Spatial analysis of spike-timing-dependent LTP and LTD in the CA1 area of hippocampal slices using optical imaging.

Spike-timing-dependent long-term potentiation (LTP) and long-term depression (LTD) were investigated in the CA1 area of hippocampal slices using optical imaging. A pair of electrical pulses were used to stimulate the Schaffer-commissural collateral and the stratum oriens with various sets of relative timing (tau) between the two stimuli. These sets of paired pulses gave rise to LTP/LTD, whose induction was closely related to tau, and their profiles were classified into two types depending on their layer specific location along the dendrite. One was characterized by a symmetric time window observed in the proximal region of the stratum radiatum (SR) and the other by an asymmetric time window in the distal region of the SR. Bath application of bicuculline (gamma-aminobutyric acid [GABA] receptor antagonist) to hippocampal slices revealed that GABAergic interneuron projections were responsible for the symmetry of a time window.

Corticosterone shifts different forms of synaptic potentiation in opposite directions

Calcium entering the cell via different routes, e.g.,N-methyl-D-aspartate (NMDA) receptors or voltage-dependent calcium channels (VDCCs), plays a pivotal role in hippocampal synaptic potentiation. Since corticosteroid hormones have been reported to enhance calcium influx through VDCCs, one may predict that these hormones facilitate hippocampal synaptic efficacy. Surprisingly, though, stress and corticosteroids have so far been found to reduce synaptic potentiation. Here, we addressed this apparent paradox and examined synaptic potentiation in the CA1 area of hippocampal slices from mice with low basal corticosterone levels 1--4 h after a brief in vitro administration of corticosterone. Nifedipine and APV were used to isolate NMDA receptor-mediated and VDCC-mediated long-term potentiations (LTPs), respectively. We report that corticosterone facilitates synaptic potentiation that depends on activation of VDCCs while impairing synaptic plasticity that is mediated by NMDA receptor activation. The glucocorticoid-receptor (GR) antagonist RU 38486 blocked both the effects of corticosterone. These results indicate that the net effect of corticosteroid hormones on synaptic plasticity is determined by the balance between different types of potentiation, a balance that may be region specific and depends on the experimental conditions. We speculate that these opposite effects on synaptic efficacy are involved in the bidirectional modulation of cognitive performance by corticosteroid hormones.

Opposite effects of lead exposure on taurine- and HFS-induced LTP in rat hippocampus

The effect of lead exposure on taurine-induced long-term potentiation (LTP TAU ) was examined and compared with high-frequency stimulation-induced one (LTP HFS ). Field excitatory postsynaptic potentials (fEPSP) and fiber volley (FV) in area CA1 of hippocampal slice were recorded in control and lead-exposed rats. In contrast to the inhibitory effects of lead exposure on LTP HFS , the amplitude of LTP TAU in the lead-exposed rats (199.3 ± 13.7%, n = 12) was significantly larger than that in controls (152.3 ± 17.0%, n = 12). It was also observed that taurine induced greater FV potentiation in lead-exposed rats (162.6 ± 9.0%, n = 10) than controls (132.1 ± 6.9%, n = 11). In addition, after a previous HFS, sequent perfusion of taurine could further increase the synaptic efficacy in lead-exposed rats. These results provide the first evidence that chronic lead exposure has opposite effects on the two types of LTP resulting from different lead toxicity sites.

Neurogranin/RC3 Enhances Long-Term Potentiation and Learning by Promoting Calcium-Mediated Signaling

In neurons, neurogranin (Ng) binds calmodulin (CaM), and its binding affinity is reduced by increasing Ca2+, phosphorylation by PKC, or oxidation by oxidants. Ng concentration in the hippocampus of adult mice varied broadly (Ng+/+, 160-370 and Ng+/-, approximately 70-230 pmol/mg); the level in Ng+/+ mice is one of the highest among all neuronal CaM-binding proteins. Among Ng+/- mice, but less apparent in Ng+/+, a significant relationship existed between their hippocampal levels of Ng and performances in the Morris water maze. Ng-/- mice performed poorly in this task; they also displayed deficits in high-frequency-induced long-term potentiation (LTP) in area CA1 of hippocampal slices, whereas low-frequency-induced long-term depression was enhanced. Thus, compared with Ng+/+ mice, the frequency-response curve of Ng-/- shifted to the right. Paired-pulse facilitation and synaptic fatigue during prolonged stimulation at 10 Hz (900 pulses) were unchanged in Ng-/- slices, indicating their normal presynaptic function. Measurements of Ca2+ transients in CA1 pyramidal neurons after weak and strong tetanic stimulations (100 Hz, 400 and 1000 msec, respectively) revealed a significantly greater intracellular Ca2+ ([Ca2+]i) response in Ng+/+ compared with Ng-/- mice, but the decay time constants did not differ. The diminished Ca2+ dynamics in Ng-/- mice are a likely cause of their decreased propensity to undergo LTP. Thus, Ng may promote a high [Ca2+]i by a "mass-action" mechanism; namely, the higher the Ng concentration, the more Ng-CaM complexes will be formed, which effectively raises [Ca2+]i at any given Ca2+ influx. This mechanism provides potent signal amplification in enhancing synaptic plasticity as well as learning and memory.

Preserved LTP and water maze learning in hyperglycaemic–hyperinsulinemic ZDF rats

Previous investigations have demonstrated that cognitive deficits as well as hippocampal dysfunctions are generated in animals presenting manifestations of Type 1 diabetes (T1D) mellitus. The present study examined whether such deficits can also be reproduced in the Zucker Diabetic Fatty (ZDF) rats after they developed symptoms of Type 2 diabetes (T2D). Learning and memory assessments were performed using the Morris water maze 5 weeks after the animals presented symptoms of Type 1 diabetes for Experiment 1 (Exp 1) and after 8 weeks for Experiment 2 (Exp 2). Testing in the water maze revealed that ZDF rats learned the task normally, although control rats were found to swim significantly faster after 5 or 8 weeks of untreated diabetes. From an electrophysiological perspective, we observed that the integrity of synaptic function was also preserved in ZDF rats as no alterations in long-term potentiation (LTP) were observed in the area CA1 of hippocampal slices. It is concluded that hyperglycaemia is not the only factor influencing water maze learning and LTP in this animal model of Type 2 diabetes (T2D). The experiments suggest that the resistance of ZDF rats to cognitive and electrophysiological dysfunctions might be related to the protective action of hyperinsulinemia. Indeed, measurements of the plasma insulin level at the end of testing were significantly superior in ZDF rats in comparison to control rats.

Alterations in synaptic transmission and plasticity in area CA1 of adult hippocampus following developmental hypothyroidism

Transient reductions in thyroid hormone during critical periods of brain development can have devastating and irreversible effects on neurological function. The hippocampus is a brain region sensitive to thyroid hormones and is a necessary substrate for some forms of learning and memory. Subregions within the hippocampus display distinct ontogenetic profiles and have shown differential vulnerability to some indices of thyrotoxic insult. Synaptic function can be readily assessed in the hippocampus, yet little information exists on the consequences of early thyroid hormone insufficiency on the neurophysiological integrity of this structure. Previous work has examined the long-term consequences of perinatal hypothyroidism on neurophysiology of the dentate gyrus of the hippocampal formation. The current study reveals that alterations in synaptic function also exist in area CA1, and some differences in the pattern of effects are evident between the two hippocampal subfields. Developing rats were transiently exposed to the thyrotoxicant, propylthiouracil (PTU; 0 or 15 ppm), through the drinking water of pregnant dams beginning on gestational day 18. This regimen markedly reduced circulating levels of thyroid hormones and stunted pup growth. PTU exposure was terminated on postnatal day (PN) 21 and electrophysiological assessments were conducted by recording field potentials in area CA1 of hippocampal slices derived from adult male offspring. Synaptic transmission, short-term, and long-term synaptic plasticity were assessed. Consistent with observations in the dentate gyrus, somatic population spike amplitudes were reduced in assessments of baseline synaptic transmission of slices from PTU-exposed animals. No differences were identified in excitatory postsynaptic potentials (EPSP). Short-term plasticity of the EPSP as indexed by paired pulse facilitation was markedly impaired by PTU exposure. Long-term potentiation (LTP) of the population spike was enhanced, consistent with findings in dentate gyrus, but no change in EPSP LTP was detected. Perturbations in synaptic function in the hippocampus of adult rats transiently exposed to a period of hormone insufficiency during the perinatal period are likely to contribute to cognitive deficits associated with developmental hypothyroidism.

The influence of developmental periods of sodium valproate exposure on synaptic plasticity in the CA1 region of rat hippocampus

The effects of chronic sodium valproate (VPA) exposure during different developmental periods on synaptic plasticity were studied in the hippocampal CA1 area. Rats were exposed to VPA (300 mg/kg per day, intraperitoneal) over different developmental intervals: (1) prenatal exposure (PR group); (2) from parturition to weaning (PW group); (3) from pregnancy day 1 to weaning (PRW group). At postnatal day 22–28, field excitatory postsynaptic potentials were recorded in the CA1 area of hippocampal slices. We found that VPA exposure in the PR and PRW groups significantly impaired long-term potentiation (LTP), long-term depression (LTD) and paired-pulse facilitation (PPF). However, exposure during lactation (PW group) only impaired LTP and had no significant effect on LTD and PPF. These results suggested that VPA exposure during lactation had the least deficits on synaptic plasticity, while VPA exposure in the PR and PRW groups seriously impaired synaptic plasticity and should be avoided.

The influence of developmental periods of sodium valproate exposure on synaptic plasticity in the CA1 region of rat hippocampus

The effects of chronic sodium valproate (VPA) exposure during different developmental periods on synaptic plasticity were studied in the hippocampal CA1 area. Rats were exposed to VPA (300 mg/kg per day, intraperitoneal) over different developmental intervals: (1) prenatal exposure (PR group); (2) from parturition to weaning (PW group); (3) from pregnancy day 1 to weaning (PRW group). At postnatal day 22–28, field excitatory postsynaptic potentials were recorded in the CA1 area of hippocampal slices. We found that VPA exposure in the PR and PRW groups significantly impaired long-term potentiation (LTP), long-term depression (LTD) and paired-pulse facilitation (PPF). However, exposure during lactation (PW group) only impaired LTP and had no significant effect on LTD and PPF. These results suggested that VPA exposure during lactation had the least deficits on synaptic plasticity, while VPA exposure in the PR and PRW groups seriously impaired synaptic plasticity and should be avoided.

Pre- and postnatal propylthiouracil-induced hypothyroidism impairs synaptic transmission and plasticity in area CA1 of the neonatal rat hippocampus.

Thyroid hormones are essential for neonatal brain development. It is well established that insufficiency of thyroid hormone during critical periods of development can impair cognitive functions. The mechanisms that underlie learning deficits in hypothyroid animals, however, are not well understood. As impairments in synaptic function are likely to contribute to cognitive deficits, the current study tested whether thyroid hormone insufficiency during development would alter quantitative characteristics of synaptic function in the hippocampus. Developing rats were exposed in utero and postnatally to 0, 3, or 10 ppm propylthiouracil (PTU), a thyroid hormone synthesis inhibitor, administered in the drinking water of dams from gestation d 6 until postnatal day (PN) 30. Excitatory postsynaptic potentials and population spikes were recorded from the stratum radiatum and the pyramidal cell layer, respectively, in area CA1 of hippocampal slices from offspring between PN21 and PN30. Baseline synaptic transmission was evaluated by comparing input-output relationships between groups. Paired-pulse facilitation, paired-pulse depression, long-term potentiation, and long-term depression were recorded to examine short- and long-term synaptic plasticity. PTU reduced thyroid hormones, reduced body weight gain, and delayed eye-opening in a dose-dependent manner. Excitatory synaptic transmission was increased by developmental exposure to PTU. Thyroid hormone insufficiency was also dose-dependently associated with a reduction paired-pulse facilitation and long-term potentiation of the excitatory postsynaptic potential and elimination of paired-pulse depression of the population spike. The results indicate that thyroid hormone insufficiency compromises the functional integrity of synaptic communication in area CA1 of developing rat hippocampus and suggest that these changes may contribute to learning deficits associated with developmental hypothyroidism.

Effects of sodium valproate on synaptic plasticity in the CA1 region of rat hippocampus

Sodium valproate (VPA) is currently one of the major anticonvulsant drug in clinical use and has a wide spectrum of antiepileptic activity. Previous studies have reported that VPA impairs long-term potentiation (LTP). In the present study, we used two forms of synaptic plasticity, LTP and long-term depression (LTD) of field excitatory postsynaptic potential (fEPSP) to investgate the effects of VPA on synaptic plasticity in rat hippocampal slices. Paired-pulse facilitation (PPF) and field EPSP were recorded in the CA1 area of hippocampal slices exposed to VPA. The results showed that: (1) three different concentrations of VPA (0.6, 1 and 5 m m) all induced a significant impairment of PPF at 20–150 ms inter-pulse intervals (IPI) ( P<0.05). (2) acute VPA exposure (0.6 m m) inhibited the induction of LTP (Control: 171±20%, n=8; VPA-exposed: 117±16%, n=9, P<0.01) and LTD (Control: 86±13%, n=8; VPA-exposed: 98±8%, n=10, P<0.01); and (3) GABA A receptor antagonist picrotoxin (PTX) (10 μM) reversed VPA-induced deficits of LTP (VPA-exposed: 117±16%, n=9; VPA-exposed +PTX: 153±20%, n=8, P<0.01). However, PTX had no significant effect on impairment of LTD (VPA-exposed: 98±8%, n=10; VPA-exposed+PTX: 97±3%, n=8, P>0.05). These results suggested that VPA impaired LTP and LTD. Furthermore, VPA-induced impairment of LTP could be correlated with the enhancement of inhibitory neurotransmission mediated by γ-aminobutyric acid (GABA) receptor.

Nicotinic receptors differentially regulate N-methyl-D-aspartate damage in acute hippocampal slices.

Although in neuronal cultures nicotine was reported to prevent early and delayed excitotoxic death, no studies with nicotinic drugs have been done with acute hippocampal slices. We investigated the effect of nicotine and methyllycaconitine (MLA) on the toxicity of N-methyl-d-aspartate (NMDA) in the CA1 area of hippocampal slices. The excitotoxic effect of NMDA was assessed as decreased recovery of the capability to produce synaptically evoked population spikes (PSs). Application of nicotine or MLA before NMDA application increased the recovery of PSs. This electrophysiological recovery was used as a measure of the early neuroprotective events. The neuroprotection conferred by both nicotine and MLA was inhibited by dihydro-beta-erythroidine, showing mediation of neuroprotection by alpha 4 beta 2 neuronal nicotinic receptors (nAChRs). Because nicotine activates alpha 4 beta 2 and other nAChR subtypes, whereas 10 nM MLA inhibits the alpha 7 subtype, we propose the involvement of a neuronal circuitry-dependent mechanism for nicotinic neuroprotection. The effect of nicotine downstream from the receptors was investigated using inhibitors of cell signaling. The results suggest that the effect of nicotine is mediated by tyrosine receptor kinases, 1,2-phosphatidylinositol-3 kinase, and the mitogen-activated extracellular signal-regulated kinases. Although nicotine neuroprotection is Ca2+-dependent, neither L-type Ca2+ channels nor calmodulin-dependent protein kinase is involved in the effect of nicotine. In summary, these results suggest that in acute slices nicotinic protection is initiated either by direct activation of alpha 4 beta 2 or indirectly by inhibition of alpha 7 followed by signal transduction involving tyrosine kinases, phospholipid-dependent kinases, and mitogen-activated kinases.

NMDA receptor regulation by amyloid-β does not account for its inhibition of LTP in rat hippocampus

Accumulation of amyloid-β peptide (Aβ) is widely believed to play a critical role in the pathogenesis of Alzheimer’s disease. Although amyloid-containing plaques are a key neuropathological feature of AD, soluble forms of Aβ can interfere with synaptic plasticity in the brain, suggesting that this form of the peptide may be responsible for much of the memory deficit seen early in the disease. Here, we investigate the mechanism underlying the effects of Aβ on long-term potentiation (LTP) in area CA1 of rat hippocampus. Extracellular field recordings were made in area CA1 of hippocampal slices taken from young, adult male rats. A non-toxic concentration of Aβ (200 nM) produced a rapid inhibition of LTP induced by 100 Hz stimulation while having no long-term effect on normal synaptic transmission. The same dose of Aβ had no effect on long-term depression (LTD) induced by 1200 pulses at 1 or 3 Hz. Picrotoxin had no effect on the inhibition of LTP, suggesting Aβ does not act by enhancing GABAergic transmission. Since the LTP induction in this study was dependent on N-methyl- d-aspartate (NMDA) receptor activation, we looked at the effect of Aβ on isolated NMDA receptor-mediated field potentials. Aβ produced a small but significant inhibition of NMDA receptor-mediated synaptic potentials (∼25%). However, a low dose of MK-801 (0.5 μM) that produced a similar inhibition of NMDA potentials had no effect on LTP induction but completely blocked LTD induction. These results suggest that Aβ does not inhibit LTP via effects on NMDA receptors, but rather interferes with a downstream pathway.