Developmental lead (Pb) exposure reduces the ability of the NMDA antagonist MK-801 to suppress long-term potentiation (LTP) in the rat dentate gyrus, in vivo.

Chronic developmental lead (Pb) exposure increases the threshold and enhances decay of long-term potentiation (LTP) in the dentate gyrus of the hippocampal formation. MK-801 and other antagonists of the N-methyl-D-aspartate (NMDA) glutamate receptor subtype impair induction of LTP. In addition, Pb exposure reduces presynaptic glutamate release and is associated with alterations in NMDA receptor expression. This study examined LTP in Pb-exposed animals challenged with a low dose of MK-801 to assess the sensitivity of this receptor to inhibition. Pregnant rats received 0.2% Pb acetate in the drinking water beginning on gestational day 16, and this regimen was continued through lactation. Adult male offspring maintained on this solution from weaning were prepared with indwelling electrodes in the perforant path and dentate gyrus. Several weeks later, input/output (I/O) functions were collected in awake animals before and after saline or MK-801 administration (0.05 mg/kg, s.c.). LTP was induced using suprathreshold train stimuli 60 min post-drug. Post-train I/O functions were reassessed 1 and 24 h after train delivery. Upon full decay of any induced LTP, drug conditions were reversed such that each animal was tested under saline and MK-801. I/O functions measured 1 and 24 h after train induction as well as immediate post-train responses revealed significant LTP of comparable magnitude that was induced in both control and Pb-exposed animals tested under saline conditions. In contrast, MK-801 reduced LTP in control but not in Pb-exposed animals. The broadening of the excitatory postsynaptic potential evident in responses evoked by train stimuli is NMDA-dependent. Pb exposure attenuated the MK-801-induced reduction in area of this NMDA component by approximately 50%. These findings are consistent with other neurochemical and behavioural observations and suggest that up-regulation of postsynaptic NMDA receptors produces subsensitivity to the inhibitory effects of MK-801 on hippocampal LTP following chronic developmental Pb exposure.

Interaction of dietary tryptophan and social isolation on territorial aggression, motor activity, and neurochemistry in mice.

This study examined the interaction of dietary tryptophan (TRP) and differential housing on territorial-induced aggression, locomotor activity, and monoamine neurochemistry in mice. Groups of male CF-1 mice were singly-housed or group-housed and administered a semisynthetic basal diet supplemented with TRP (0.25-1.0%). Behavioral measures were taken at various intervals up to 2 weeks after dietary administration was instituted. Separate groups of mice were given the same experimental treatment and sacrificed for whole brain determination of the monoamines and their metabolites. Isolated mice were consistently more aggressive than grouped animals, suggesting that territorial-induced aggression is synergistic with intermale aggression based on social isolation. The combination of isolation and 0.50% TRP was particularly effective in producing increases in aggression that reached maximal levels after 10 days of diet administration. However, motor activity of singly-housed mice was unaffected by TRP, while that of grouped mice was decreased after 5 days of 0.50% TRP. By day 14 of administration behavioral changes tended to return to baseline levels. Neurochemical measures indicated increased DA and 5-HT turnover in isolated mice, with the 5-HT system most affected by dietary TRP. Because housing conditions were a prominent factor in the aggression and neurochemistry, the results suggest the involvement of both transmitter systems in this behavior. However, there were no changes in monoamine turnover that could account for the development of behavioral tolerance.

Differential tolerance to dietary amino acid-induced changes in aggressive behavior and locomotor activity in mice.

Male albino mice were maintained on a semisynthetic 12% casein protein diet for 2 weeks, then switched to diets modified by the addition of a 4% L-amino acid supplement (L-tyrosine, L-phenylalanine, and L-tryptophan) or 4% casein (control). Territorial=induced aggressive behavior increased following 1 week on the amino acid supplements, especially after tyrosine, but an apparent tolerance developed to these effects after 5 weeks on the amino acid supplements. Locomotor activity also increased following 1 week on the supplements, most notably after phenylalanine alone or in combination with tyrosine, and these effects tended to persist after 5 weeks on the supplements. Endogenous whole brain levels of dopamine, norepinephrine, serotonin, 5-hydroxyindoleacetic acid, tyrosine, phenylalanine, and tryptophan showed no tolerance to increased concentrations of brain catecholamines and indoleamines over the 5-week period, and no clear relation between the concentrations of these monoamines and the behavioral changes.

Rat hippocampal glutamate and GABA release exhibit biphasic effects as a function of chronic lead exposure level.

Previous work has suggested that the lead (Pb) exposure-induced decrease in K(+)-evoked hippocampal glutamate (GLU) release is an important factor in the elevated threshold and diminished magnitude reported for hippocampal long-term potentiation (LTP) in exposed animals. In addition, complex dose-effect relationships between Pb exposure level and LTP have been reported. This investigation was conducted to determine the effects of Pb on hippocampal GLU and GABA release as a function of exposure level. Rats were continuously exposed to 0.1, 0.2, 0.5, or 1.0% Pb in the drinking water beginning at gestational day 15-16. Hippocampal transmitter release was induced in adult males by perfusion of 150 mM K(+) in the presence of Ca(+2) (total release) through a microdialysis probe in one test session, followed by perfusion through a contralateral probe in the absence of Ca(+2) (Ca(+2)-independent release) in the second session. Chronic exposure produced decreases in total K(+)-stimulated hippocampal GLU and GABA release at exposure levels of 0.1-0.5% Pb. Maximal effects were seen in the 0.2% group (blood Pb = 40 microg/100 ml), and changes in total release could be directly traced to alterations in the Ca(+2)-dependent component. However, these effects were less evident in the 0.5% group and were no longer present in the 1.0% Pb group, thus defining U-shaped dose-effect relationships. Moreover, in the absence of Ca(+2) in the dialysis perfusate, K(+)-induced release was elevated in the 2 highest exposure groups, suggesting a Pb(+2)-induced enhancement in evoked release. This pattern of results indicates the presence of 2 actions of Pb on in vivo transmitter release: a more potent suppression of stimulated release seen at lower exposure levels (27-62 microg/100 ml) combined with Ca+2-mimetic actions to independently induce exocytosis that is exhibited at higher exposure levels (> or =62 microg/100 ml). Furthermore, significant similarities in the dose-effect relationships uncovered in measures of evoked GLU release and hippocampal LTP (M. E. Gilbert et al., 1999b, Neurotoxicology 20, 71-82) reinforce the conclusion that exposure-related changes in GLU release play a significant role in the Pb-induced effects seen in this model of synaptic plasticity.

Bridging the gap between in vitro and in vivo models for neurotoxicology.

In vitro systems are widely used for investigation of neurotoxicant-induced perturbations of cellular functions. A variety of systems exist that demonstrate certain similarities to neurotoxicant-induced events in the intact animal are discussed, including single-cell types, systems that consider endpoints relevant in toxicology, and systems that consider heterogeneous cell interactions. Relationships between the in vitro and in vivo systems are examined in which ethanol, lead, polychlorinated biphenyl compounds, and organophosphate insecticides are examples. Situations in which the in vitro systems have been used to advantage are provided, along with cautions associated with their use.

The combined EEG-intracerebral microdialysis technique: a new tool for neuropharmacological studies on freely behaving animals.

In this study we combined EEG and intracerebral microdialysis techniques in freely behaving rats. Various drugs were delivered into the hippocampus and cerebral cortex by means of microdialysis and, simultaneously, the EEG activity of the dialyzed area was monitored. The microdialysis procedure itself, when artificial cerebrospinal fluid was perfused, did not change the normal hippocampal or cortical EEG pattern. Drug inclusions into the microdialysis fluid, however, caused marked changes in the electrical activity of the dialyzed sites. In this report we present the following examples: (1) the dose-dependent spike-provoking effect of NMDA in hippocampus, (2) the potentiation of this NMDA effect in hippocampus by dibutyryl cyclic AMP, and (3) the EEG depressant effect of high concentration of K+ in the cerebral cortex. The artificial cerebrospinal fluid and drug solutions were alternated in the microdialysis system with a 2-way valve placed outside the test chamber. As a consequence, the drugs were delivered into the brain without interrupting the ongoing behavior, including sleep, of the examined animals. This study shows that the combined EEG-intracerebral microdialysis technique is a useful tool, with many unique advantages, for in vivo neuropharmacological studies.

Roles of neurotransmitter amino acids in seizure severity and experience in the genetically epilepsy-prone rat.

This investigation was designed to compare seizure-naive and seizure-experienced genetically epilepsy-prone rats (GEPRs) in order to distinguish transmitter amino acid changes related to seizure severity from those associated with seizure experience. Moderate (GEPR-3) and severe (GEPR-9) seizure male GEPRs were divided into seizure-naive and seizure-experienced groups based on whether seizure-inducing acoustical stimuli had been presented between 45 and 60 days of age, and then were sacrificed at 76 +/- 3 days. gamma-Aminobutyric acid (GABA) concentrations were lower in both GEPR-3s and GEPR-9s compared to non-epileptic controls in each brain region examined. Aspartate content was elevated in 5 of 6 brain areas in GEPR-9s compared to non-epileptic controls, and in 3 regions was higher in GEPR-9s than in GEPR-3s. In contrast, taurine concentrations were higher in GEPR-3s than in non-epileptic controls in each region, and in 4 areas were higher in GEPR-3s than in GEPR-9s. Changes resulting from seizure experience consisted of increases in aspartate, glutamate and glycine in seizure-experienced compared to seizure-naive groups in inferior colliculus and in motor-sensory and frontal cortices. These findings suggest that the high levels of taurine in GEPR-3s and the elevated content of aspartate in GEPR-9s have roles as determinants of seizure severity. The low concentrations of GABA in both types of GEPRs are consistent with a role for this amino acid in determination of seizure susceptibility. Furthermore, the seizure-induced changes in aspartate and glutamate in both types of GEPRs support the concept that these excitatory amino acids mediate changes in seizure predisposition.(ABSTRACT TRUNCATED AT 250 WORDS)

Presynaptic glutamatergic function in dentate gyrus in vivo is diminished by chronic exposure to inorganic lead.

Reductions in membrane Ca2+ channel currents and depolarization-evoked neurotransmitter release have been repeatedly observed as a result of acute exposure to Pb2+. This study was performed to determine whether hippocampal glutamate and GABA release are impaired in intact animals chronically exposed to lead (Pb). As paired-pulse facilitation in the hippocampus is primarily mediated by an enhancement of glutamate release, this neurophysiological measure was also assessed in the dentate gyrus of Pb-exposed animals. Pregnant dams received 0.2% Pb acetate in the drinking water at parturition, and male offspring were weaned to the same solution as that given their dams. Control animals were maintained on distilled water. As adults, animals had intracerebral dialysis probes inserted through guide cannulae implanted 2-4 days previously and the hippocampal CAI-dentate area was perfused with modified Ringer's solution. Transmitter release was induced by perfusion with 150 mM K+ with half the animals in each group tested with Ca2+ present in the perfusate (total release) and the other half with Ca2+ absent (Ca(2+)-independent release). K(+)-stimulated total glutamate release was reduced in Pb-exposed animals relative to controls. No group differences were observed under Ca(2+)-free conditions, indicating that Ca(2+)-dependent glutamate release was decreased in exposed rats. In contrast no group differences in K(+)-stimulated total GABA release were evident, whereas an augmentation in GABA release under Ca(2+)-free conditions was revealed in Pb-exposed animals. The effects of exposure on the Ca(2+)-dependent components of release are consistent with in vitro evidence indicating an inhibitory action of Pb2+ at voltage-sensitive Ca2+ channels. A separate group of animals was prepared under urethane anesthesia with stimulating and recording electrodes placed in the perforant path and dentate gyrus, respectively. Pairs of stimulus pulses were delivered at interpulse intervals (IPI) of 10-250 ms. Pb exposure induced an increase in paired-pulse depression at the 20 ms 1PI and reduced paired-pulse facilitation at the 30 ms IPI. Decreases in paired-pulse facilitation could not be attributed to the reported effects of Pb2+ on N-methyl-D-aspartate (NMDA) receptors as MK-801 (1.0 mg/kg, s.c.) administration produced an opposing pattern of effects on paired-pulse measures. The Pb-induced suppression of paired-pulse facilitation is consistent with exposure-related decreases in total glutamate release. The impact of these effects of Pb exposure on hippocampal glutamatergic transmission may contribute to the reported effects of Pb on other forms of synaptic plasticity including long-term potentiation, a model of learning and memory.

Chronic exposure to environmental levels of lead impairs in vivo induction of long-term potentiation in rat hippocampal dentate.

This study examined the effects of chronic developmental lead (Pb) exposure in rats on hippocampal long-term potentiation (LTP). Male offspring were exposed to 0.2% Pb acetate continuously from birth until testing at 85-105 days. Excitatory postsynaptic potential (EPSP) and population spike amplitudes were measured in the dentate hilar region in response to stimulation applied to the lateral perforant path. LTP was induced in control animals with an average maximal EPSP potentiation of 41%, which was significantly greater than the increase in EPSP amplitudes (2%) in exposed animals after tetanizing stimulation. Current-voltage curves in controls demonstrated significant increases in EPSPs and population spikes after application of pulse trains to induce LTP, while exposed rats exhibited no discernible change in responses. These findings suggest that induction or development of LTP in the dentate hilar region in vivo is impaired by chronic developmental exposure to environmentally relevant levels of Pb.

Chronic developmental lead exposure increases the threshold for long-term potentiation in rat dentate gyrus in vivo.

Chronic developmental lead (Pb) exposure has been long associated with cognitive dysfunction in children and animals. In an attempt to more directly relate the behavioral observations of impaired cognitive ability to Pb-induced effects on neuronal activity, we utilized the long-term potentiation (LTP) model of neural plasticity to assess synaptic function. Male rats were chronically exposed to 0.2% Pb(2+)-acetate through the drinking water of the pregnant dam, and directly through their own water supply at weaning. As adults, field potentials evoked by perforant path stimulation were recorded in the dentate gyrus under urethane anesthesia. LTP threshold was determined by applying a series of stimulus trains of increasing intensities. Baseline testing of dentate gyrus field potentials indicated that input/output functions, maximal response amplitudes, and threshold currents required to evoke a population spike (PS) did not differ for control and Pb-exposed animals. Despite similarities in baseline synaptic transmission, Pb-exposed animals required a higher train intensity to evoke LTP than controls. With maximal train stimulation, however, control and Pb animals exhibited comparable levels of potentiation. These findings suggest that the mechanisms of LTP induction are preferentially impaired by Pb exposure. Although baseline synaptic transmission was not altered in Pb-exposed animals, decreases in glutamate release following high K+ perfusion and reductions in paired pulse facilitation have been reported in the intact animal. Pb-induced reductions in calcium influx through voltage-sensitive or N-methyl-D-aspartate (NMDA) receptor-dependent channels may mediate increases in LTP threshold. It is possible that the threshold changes in the induction of LTP reported here contribute to cognitive impairments associated with Pb exposure.

Diminished potassium-stimulated GABA release in vivo in genetically epilepsy-prone rats.

This experiment was conducted to assess the physiological relevance of observed changes in transmitter amino acid content in severe seizure genetically epilepsy-prone rats (GEPR-9s) by use of microdialysis. Adult male GEPR-9s and non-epileptic control rats were implanted with guide cannulae, and 6 mm (loop) dialysis probes were inserted unilaterally into rostral caudate and perfused with artificial cerebrospinal fluid. Each subject was perfused in the awake state with 100 or 150 mM K+ for 80 min in separate counterbalanced sessions, and 20-min fractions collected. High K+ perfusion increased extracellular fluid GABA and glutamate (GLU) in a concentration-dependent manner in both GEPR-9s and non-epileptic control rats. However, in the presence of 150 mM K+ GABA release was decreased in GEPR-9s relative to controls throughout the stimulation interval. In contrast, the increase in extracellular fluid GLU after high K+ was not different in the two groups. These results suggest an important role for mechanisms underlying GABA release in the seizure susceptibility observed in GEPR-9s.

Amino acids, monoamines and audiogenic seizures in genetically epilepsy-prone rats: effects of aspartame.

It has been suggested that aspartame facilitates seizures in man and animals because phenylalanine, one of its major metabolites, interferes with brain transport of neurotransmitter precursors and alters the synthesis of monoamine neurotransmitters such as norepinephrine, dopamine and/or serotonin. This facilitation is purportedly more likely in subjects predisposed to seizures. One test of this hypothesis would be to administer a wide range of aspartame doses to subjects whose seizure predisposition is dependent on abnormalities in monoaminergic function. Genetically epilepsy-prone rats (GEPRs) have a broadly based seizure predisposition that is based, in part, on widespread central nervous system noradrenergic and serotonergic deficits. Further reductions in the functional state of these neurotransmitters increases seizure severity in GEPRs. Thus, GEPRs appear ideally suited for testing the hypothesis that aspartame facilitates seizures by interfering with central nervous system monoamines. Oral administration of acute (50-2000 mg/kg) or sub-chronic (up to 863 mg/kg/day for 28 days) doses of aspartame did not alter seizure severity in either of two types of GEPRs. Not surprisingly, acute aspartame doses produced dramatic changes in plasma and brain amino acid concentrations. Hypothesized alterations in monoamine neurotransmitter systems were largely absent. Indeed, increases in norepinephrine concentration, rather than the hypothesized decreases, were the most evident alterations in these neurotransmitter systems. We conclude that aspartame does not facilitate seizures in GEPRs and that convincing evidence of seizure facilitation in any species is lacking.

Regulation of dopaminergic activity, but not tyrosine hydroxylase, is diminished after chronic inorganic lead exposure.

Previous work has indicated that the neurotoxic action of environmentally relevant levels of lead (Pb) on dopaminergic neurons is primarily presynaptic in nature and related to impaired regulation of dopamine (DA) synthesis and decreased DA release. This study was conducted to assess the functional integrity of the regulation of DA synthesis in caudate-putamen (C-P) and nucleus accumbens (NAc) of chronically Pb-exposed rats by measuring tyrosine hydroxylase (TH) activity. A pharmacological paradigm was employed that isolated autoreceptor-mediated regulation of the enzyme. At parturition dams received 0.2% Pb acetate (1090 ppm) in the drinking water while control dams received distilled water. Offspring were weaned to and maintained on the same solution given their dams until termination at 60 or 120 days. Rats were given saline or one of three DA agonists (EMD 23448, CGS 15855A, TL-99) 45 or 60 min before termination followed 15 min later by 750 mg/kg i.p. of gamma-butyrolactone (GBL) or saline. The ability of a DA agonist to prevent the GBL-induced increase in DA content was significantly altered in C-P of exposed rats compared to controls. No effect of Pb on DA content was observed in NAc. Furthermore, no differences in the ability of DA agonists to inhibit GBL-induced activation of TH in Pb-exposed compared to control animals were apparent in either brain region at either age by use of the tritium release method or the accumulation of L-DOPA. On the other hand, concentrations of DA metabolites in exposed rats given GBL and EMD 23448 were significantly lower than those in controls in both C-P and NAc. These findings support previous work suggesting that chronic Pb has multiple actions on CNS dopaminergic neurons consisting of impaired regulation of DA content and decreased DA release. However, these effects cannot be attributed to alterations in autoreceptor-mediated regulation of TH activity.

Glutamatergic components underlying lead-induced impairments in hippocampal synaptic plasticity.

Epidemiological investigations have established the relationship between chronic developmental lead (Pb) exposure and cognitive impairments in young children, defining Pb neurotoxicity as a significant pediatric health problem. Exposed animals have proven to be effective models of this condition, exhibiting similar sensitivity to the actions of Pb and replicating abnormal learning behaviors in exposed children. Research has extended these observations in animals to identifying the processes underlying the cognitive dysfunction, utilizing the long-term potentiation (LTP) paradigm as a correlate of learning ability. Results from these studies have been in widespread agreement in reporting impairments in synaptic plasticity. Exposure-related changes consist of increases in LTP induction threshold, decreases in magnitude of potentiation, and shortened LTP duration. Furthermore, while LTP may be more readily affected by Pb during early development, exposure initiated after weaning also potently affects synaptic plasticity. Biphasic dose-effect relationships also appear in which impaired LTP is observed at intermediate exposure levels (27-62 microg/100 ml), but not at higher exposures. Investigation of the synaptic processes underlying LTP has provided additional insight into the bases of the impaired potentiation and diminished cognitive ability. Biochemical and neurophysiological approaches have found stimulated glutamate release to be diminished in hippocampus at blood Pb values where deficits in LTP have been observed. Multiple actions of Pb may be involved at this exposure level since animals exposed postweaning exhibited similar decrements in evoked glutamate release to those exposed continuously from conception, similar to the observations in measures of LTP. A biphasic dose-effect relationship was also found in which stimulated glutamate release in hippocampus was decreased at intermediate exposures, but not at higher levels. A direct inhibitory effect of Pb2+ on NMDA receptor function does not appear to occur at environmentally relevant exposure levels, but both exposure-induced increases and decreases in receptor density have been reported by different workers. Evidence from behavioral and neurophysiological investigations can be explained by increased NMDA receptor density on the bases of increased sensitivity to agonists and decreased sensitivity to antagonists. From this body of findings it is apparent that decreases in stimulated glutamate release are a significant contributing factor to the exposure-related changes seen in LTP. Furthermore, despite general agreement on the actions of Pb on synaptic plasticity, reports of exposure effects on NMDA receptor function have been relatively variable, suggesting either that the nature of the receptor changes are dependent on exposure conditions or that the receptors are secondarily affected by Pb actions produced at signal transduction or cellular loci.

Influence of exposure period on in vivo hippocampal glutamate and GABA release in rats chronically exposed to lead.

Previous work has demonstrated that continual exposure to 0.2% lead (Pb) beginning at birth diminishes depolarization-induced hippocampal glutamate (GLU) and GABA release in vivo. The present study sought to extend these findings by examining Pb-induced changes as a function of exposure period. Rats were continually exposed to 0.2% Pb in the drinking water beginning at conception (Gestational-Life, GL) or two weeks after weaning (Wean-Life, WL), while exposure in a third group was begun at conception but terminated at weaning (Gestational-Wean, GW). Hippocampal transmitter release was induced in adult animals by perfusion of 150 mM K+ in the presence of Ca+2 (total release) through a microdialysis probe in one test session, followed by perfusion through a contralateral probe in the absence of Ca+2 (Ca+2-independent release) in the second session. Decreases in total GLU and GABA release were observed in the GL and WL groups compared to controls over the first 20-min after initiation of high K+, decrements that could be attributed to exposure-induced reductions in Ca+2-dependent release. The pattern of Pb-induced changes in the GL group is similar to that observed previously in a group continuously exposed from birth, indicating that gestational exposure did not further enhance the impact of Pb beginning at birth when exposure in both groups extends into adulthood. Similar responses were also found in the WL group, indicating that exposure during early development is not a requirement to induce changes in GLU and GABA release. Pb-induced decreases in response were also seen in the GW group: a decrease in Ca+2-dependent GLU release was observed, while decrements in total and Ca+2-dependent GABA release were similar to those in the GL and WL groups. Thus, exposure limited to early development is also sufficient to produce deficits in evoked transmitter release. In addition, the exposure-induced decreases in GLU responses correspond to Pb-induced impairments in long-term potentiation (LTP) observed in similarly exposed groups (Gilbert et al., 1999), providing further evidence that Pb effects on GLU release are a critical factor in the alterations found in LTP.

The influence of developmental period of lead exposure on long-term potentiation in the adult rat dentate gyrus in vivo.

Previous work has demonstrated an increase in the threshold for induction of long-term potentiation (LTP) in the dentate gyrus of animals chronically exposed to lead (Pb) from birth (Gilbert et al., 1996). The present study sought to extend these findings by evaluating the developmental periods critical for Pb-induced impairment of LTP. Rats were exposed to Pb through maternal milk and/or the drinking water over different developmental intervals: 1) beginning just prior to birth and continuing throughout life (PL); 2) beginning just prior to birth and terminating at weaning (PW); or 3) continuously from the early post-weaning period throughout life (WL). Pregnant dams received 0.2% Pb-acetate in the drinking water on gestational day (GD)16, with male offspring switched to the same solution (PL group) or tap water (PW group) at weaning on postnatal day (PND)21. Postweaning exposure began on PND30 and continued throughout life. As adults (PND130-210), field potentials evoked by perforant path stimulation were recorded in the dentate gyrus under urethane anesthesia, and an ascending series of stimulus trains was administered to induce LTP and to determine its threshold. The magnitude of population spike (PS) LTP was reduced relative to controls in animals exposed throughout life (PL) and in animals exposed after weaning (WL). No impairment in PS LTP was evident in animals removed from Pb at weaning and tested as adults (PW). Similarly, thresholds for induction of PS LTP were elevated relative to controls in the PL and WL groups, but were not affected by Pb exposure limited to the lactational period (PW). Reductions in the magnitude of LTP of the EPSP slope were evident in posttrain I/O functions in all Pb-exposed groups, including the PW group. An elevated LTP threshold was evident in the EPSP slope measure in the continuously exposed group (PL) only. Thus Pb exposure restricted to the lactational period appeared less disruptive to adult LTP in the dentate gyrus than continuous exposure beginning around birth or weaning. However, EPSP slope LTP was impaired in animals exposed to Pb for as little as 30 days in the early postnatal period. An attenuated ability to support neuroplastic change in synaptic function may contribute to cognitive deficits associated with Pb-induced toxicity.

Chronic developmental lead exposure and hippocampal long-term potentiation: biphasic dose-response relationship.

Developmental exposure to lead (Pb) has long been associated with reductions in intellectual function in children and behavioral impairments in animal models of learning and memory. We have used long-term potentiation (LTP) in the dentate gyrus of the Pb-exposed rats to determine the potential of a reduced capacity for synaptic plasticity to contribute to Pb-induced cognitive dysfunction. Previous work demonstrated that developmental exposure resulting in moderate blood concentrations of Pb increase the threshold for induction of long-term potentiation (LTP) in the dentate gyrus in vivo. These findings were also suggestive of reductions in LTP magnitude (Gilbert et al., 1996). The present study was designed to further examine the effects of Pb on LTP magnitude and to determine if lower blood Pb levels commonly encountered in children are also effective in impairing synaptic plasticity in this rodent model. Pregnant dams were exposed to control tap water or 0.1, 0.2, 0.5 or 1.0% Pb-acetate in the drinking water beginning just prior to parturition (gestational day 16, GD16). Male offspring were weaned at 21 days of age (PN21) to the same solution given their dams and continued on this regimen until testing. As adults, animals were anesthetized with urethane and stimulating and recording electrodes placed in the perforant path and dentate gyrus, respectively. Post-train I/O functions taken 1 hour after delivery of a series of six high frequency (400Hz) trains revealed a reduced capacity for LTP of the PS amplitude and EPSP slope in Pb-exposed animals in all but the 1.0% group, indicative of a biphasic dose-effect relationship. The 1.0% Pb exposure was clearly less effective than the lower exposure levels in reducing LTP magnitude, and did not differ significantly from control values. The mechanisms underlying the reduced efficacy of higher exposure levels of Pb to impair LTP are not clear. Blood (26-117 microg/dl) and brain (220-1812 ng/g tissue) concentrations of Pb were elevated as a function of increasing exposure (0.1%-1.0%) and cannot readily account for the lack of an effect in the 1.0% group on LTP. We have observed a similar profile in hippocampal glutamate release employing a similar range of exposure levels, i.e., reduction of glutamate release that is absent at higher concentrations of Pb in the drinking water (Lasley et al., 1998). These and previously reported data suggest that the ability of Pb to diminish presynaptic transmitter release contributes to a reduced capacity for LTP at lower exposure levels. The reversal of the effect of Pb on glutamate release that accompanies higher exposure levels may serve to compensate for the mechanism underlying the LTP impairment and form the basis for the biphasic dose-response pattern seen with chronic developmental exposure.

Determination of gamma-aminobutyric and glutamic acids in rat brain by liquid chromatography with electrochemical detection.

The determination of glutamic and gamma-aminobutyric acids in rat brain regions by derivatization with o-phthaldialdehyde-thiol, isocratic separation by liquid chromatography, and quantification by electrochemical detection provides a simple and precise method for assessing changes in glutamatergic and GABAergic neuronal systems. Gamma-aminobutyric acid was eluted in 30-35 minutes followed by a washout step with 90% methanol to remove all amino acid derivatives with longer retention times. Homoserine was used as an internal standard. Significant increases in gamma-aminobutyric acid content in nucleus accumbens and substantia nigra could be detected 20 minutes after injection of 400 mg/kg valproic acid.

Effects of acrylamide on locomotion and central monoamine function in the rat.

Male rats receiving acrylamide (ACR) in their drinking water (100 ppm) for a six-week period displayed increased psychomotor stimulation to d-amphetamine (d-A; 1.0 mg/kg SC) under several conditions of handling and drug administration. Following behavioral tests a subset of the animals was sacrificed at 15, 50, 80 and 120 minutes following d-A and the brains removed and dissected for determinations of regional brain levels of several monoamine neurotransmitters and metabolites. ACR rats had elevated levels of 5-hydroxyindoleacetic acid (5-HIAA) in the striatum, septal area, and thalamus. The effect was most pronounced at 15 minutes post-drug with ACR rats not demonstrating a depression in 5-HIAA levels present in controls. Increases in accumben's dopamine and norepinephrine levels, evident after d-A, were of lesser magnitude in ACR-exposed rats. Decreases in dihydroxyphenylacetic acid and homovanillic acid, also evident after d-A, persisted for a longer duration in ACR-exposed rats. Light and electron microscopy of spinal cord, striatum, nucleus accumbens and thalamus did not reveal morphologic abnormalities. Sciatic nerves showed histopathological changes characteristic of multi-focal dying-back peripheral nerve degeneration. It was concluded that acrylamide's effect on the psychomotor stimulant properties of d-A may be related to changes in a serotonergic inhibitory system.

Effects of dietary tyrosine, phenylalanine, and tryptophan on aggression in mice.

Dietary amino acid regimens designed to enhance catecholaminergic and serotonergic functioning were found to differentially affect territorial-induced attacks in mice. Male albino mice were maintained on a semi-synthetic 12% casein protein diet for 2 weeks, then switched to diets modified by the addition of a 4% L-amino acid supplement, or 4% casein (control). Measures of aggressive behavior and open-field locomotor activity were obtained before and after the dietary supplements were administered. Resident mice fed supplements of L-tyrosine displayed a marked increase in the number of attacks on intruders and shorter attack latencies, but their locomotor activity was unaffected. L-phenylalanine supplements alone or in combination with L-tyrosine reduced the latency to attack and increased motility but did not affect the number of attacks. As a whole, the group of animals fed L-tryptophan showed no changes in aggression or motility.