Acetylcholine Release In Hippocampus Research Articles

Age‐related modifications of effects of ketamine and propofol on rat hippocampal acetylcholine release studied by in vivo brain microdialysis

We sometimes encounter impairment of learning and memory after general anesthesia in elderly patients. The aim of this study was to examine age-related modifications of the effects of ketamine and propofol on rat hippocampal acetylcholine (ACh) release because hippocampal cholinergic neurons are supposed to be involved in learning and memory. The experiments were performed on male Wistar young rats (2 months old) and old rats (18 months old), using in vivo brain microdialysis technique under freely moving condition. After initial sampling of three collections, test drugs were administered. The ACh release was determined by the HPLC-ECD method. In old rats, the hippocampal basal ACh release was significantly lower than in young rats. Ketamine (25 and 50 mg kg(-1) i.p.) increased and propofol (25 and 50 mg kg(-1) i.p.) decreased the hippocampal ACh release in both young and old rats. Furthermore, ketamine 50 mg kg(-1) i.p. (anesthetic dose) produced facilitatory effects on the hippocampal ACh release in young rats (193% of the basal release), while in old rats the same dose of ketamine i.p. produced more pronounced facilitatory effects on the hippocampal ACh release (317% of the basal release). On the other hand, propofol 50 mg kg(-1) i.p. (anesthetic dose) produced inhibitory effects on the hippocampal ACh release in young rats (56% of control) and in old rats (77% of control). Although the maximal inhibitory peak effects of propofol 50 mg kg(-1) i.p. did not differ significantly between young rats and old rats, decrease of the hippocampal ACh release in old rats persisted longer than in young rats. Ketamine produced more pronounced facilitatory effects on the hippocampal ACh release in old rats, as compared with young rats. On the other hand, propofol has inhibitory effects on the hippocampal ACh release in young and old rats. The aging process may suppress the ability to recover from the inhibitory anesthetic state induced by propofol.

Honokiol and magnolol increased hippocampal acetylcholine release in freely-moving rats.

Honokiol and magnolol, phenolic compounds isolated from the stem bark of Magnolia officinalis, have been demonstrated to increase choline acetyltransferase activity, inhibit acetylcholinesterase, promote potassium-induced acetylcholine release and exhibit neurotrophic function in in vitro studies. The objective of the present study was to determine the effect of these compounds on hippocampal acetylcholine release in conscious, freely-moving rats. 10(-4) M-10(-6) M of honokiol or magnolol was perfused into rat hippocampus via a dialysis probe. The results showed that at 10(-4) M concentration, honokiol and magnolol markedly increased extracellular acetylcholine release to 165.5+/-5.78% and 237.83+/-9.47% of the basal level, respectively. However, lower concentrations of either compounds failed to elicit significant acetylcholine release. This result suggests that a high dose of honokiol or magnolol may enhance in vivo hippocampal acetylcholine release.

Pregnenolone sulfate increases hippocampal acetylcholine release and spatial recognition

The pregnenolone sulfate is a neurosteroid with promnesic properties. Recently, a correlation between endogenous levels of pregnenolone sulfate in the hippocampus and performance in a spatial memory task has been reported in aged rats. Cholinergic transmission is known to modulate memory processes and to be altered with age. In the present experiment we investigated the effect of increasing doses of pregnenolone sulfate on hippocampal acetylcholine release. Our results show that intracerebroventricular administrations of this neurosteroid induced a dose-dependent increase in acetylcholine release. Administration of 12 and 48 nmol of pregnenolone sulfate induced a short lasting (20 min) enhancement of acetylcholine output with a maximum around 120% over baseline and the administration of 96 and 192 nmol doses induced a long-lasting (80 min) increase that peaked around 300% over baseline. In a second experiment we have observed that the 12 nmol dose enhanced spatial memory performance, whereas the 192 nmol dose was inefficient. These results are consistent with previous work suggesting that, a modest increase in acetylcholine release facilitates memory processes, while elevation beyond an optimal level is ineffective. Nevertheless, neurosteroids may be of value for reinforcing depressed cholinergic transmission in certain age-related memory disorders.

Long-term voluntary ethanol consumption affects neither spatial nor passive avoidance learning, nor hippocampal acetylcholine release in alcohol-preferring rats

Long-term ethanol consumption in humans and laboratory animals is associated with morphological and functional alterations of brain structures involved in cognitive processes. In the present experiments, we assessed whether voluntary long-term consumption of ethanol by alcohol-preferring (sP) rats under free choice condition with water (also) caused alterations in memory performance and hippocampal acetylcholine (ACh) release in vivo. A group of sP rats were offered a 10% v/v ethanol solution in a free choice with water for 36 weeks; controls had only tap water available. After withdrawal of ethanol, rats were tested in one trial passive avoidance test and thereafter were trained in a food-reinforced radial arm maze task for 12 days. One day after the last session in the radial-arm maze, rats were implanted with a microdialysis probe in the dorsal hippocampus and dialysate concentrations of ACh were measured. No significant differences were observed between sP drinking and control rats in retention latencies in the passive avoidance test, in radial arm-maze performance or in basal levels of hippocampal ACh release. These results show that long-term ethanol consumption by sP rats is not associated with cognitive impairments or with alterations in the hippocampal cholinergic function. To the extent that chronic ethanol intoxication can be considered a causal factor in the development of memory and neurochemical alterations, these results suggest that sP rats self-regulate ethanol consumption so as to avoid intoxication. These findings may challenge the notion that sP rat lines can be considered a valid model of human alcoholism.

Pharmacological characterization of nicotine-induced acetylcholine release in the rat hippocampus in vivo: evidence for a permissive dopamine synapse.

In this study, the mechanism of nicotine-induced hippocampal acetylcholine (ACh) release in awake, freely moving rats was examined using in vivo microdialysis. Systemic administration of nicotine (0.4 mg kg(-1), s.c.) increased the levels of ACh in hippocampal dialysates. The nicotine-induced hippocampal ACh release was sensitive to the pretreatment of neuronal nicotinic acetylcholine receptor (nAChR) antagonists mecamylamine (3.0 mg kg(-1), s.c.) and dihydro-beta-erythrodine (DHbetaE; 4.0 mg kg(-1), s.c.) as well as systemic administration of the dopamine (DA) D1 receptor antagonist SCH-23390 (R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-benzaz epine; 0.3 mg kg(-1), s.c.). Local perfusion of mecamylamine (100 microM), DHbetaE (100 microM) or SCH-23390 (10 microM) through microdialysis probe did not increase basal hippocampal ACh release. Hippocampal ACh release elicited by systemic administration of nicotine (0.4 mg kg(-1), s.c.) was antagonized by local perfusion of SCH-23390 (10 microM), but not by MEC (100 microM) or DHbetaE (100 microM). Direct perfusion of nicotine (1 mM, but not 0.1 mM) increased hippocampal ACh levels; however, this effect was relatively insensitive to blockade by co-perfusion of either mecamylamine (100 microM) or SCH-23390 (10 microM). These results suggest that nicotine-induced hippocampal ACh release occurs by two distinct mechanisms: (1) activation of nAChRs outside the hippocampus leading to DA release and subsequent ACh release involving a permissive DA synapse, and (2) direct action of nicotine within the hippocampus leading to ACh release via non-DA-ergic mechanism.

Reversal by flunarizine of the decrease in hippocampal acetylcholine release in pentylenetetrazole-kindled rats

The aim of our study was to evaluate the effect of the non-selective calcium antagonist flunarizine on hippocampal acetylcholine (ACh) release with the microdialysis technique in freely moving rats after long-term concomitant administration of pentylenetetrazole (PTZ) in comparison with rats treated long-term with PTZ (kindled animals). The basal extracellular concentration of ACh in the hippocampus of rats treated with PTZ alone was significantly reduced relative to that of vehicle-treated rats (2.04 ± 0.2 vs 3.94 ± 0.3 pmol per 20-min sample; P < 0.01). Administration of flunarizine (7.5 mg/kg i.p.) before each PTZ injection prevented this decrease in basal ACh output (3.75 ± 0.4 pmol per 20-min sample). On the contrary, the expression of PTZ-induced kindling was not prevented by administration of flunarizine. The specific antagonistic effect of flunarizine on the kindling-induced decrease in hippocampal ACh release is shared by the selective antagonist of the L-type calcium channel, nifedipine, but not by the dopamine D 2 antagonist, (−)-sulpiride, suggesting that the decrease in Ca 2+ overload by a blockade of the L-type calcium channel may be responsible for the protective action on cholinergic neurons exerted by flunarizine. These data also suggest a potential therapeutic role for flunarizine in counteracting impairment of hippocampal cholinergic activity.

Cholinergic systems and long-term potentiation in memory-impaired apolipoprotein E-deficient mice

Impairments in cholinergic neurotransmitter systems of the basal forebrain are a hallmark of Alzheimer's disease pathophysiology. The presence of the ε4 allele of apolipoprotein E was recently implicated as a major risk factor in both familial and sporadic Alzheimer's disease. The present study examined the integrity of cholinergic and non-cholinergic systems in apolipoprotein E-deficient, memory-impaired mice. Choline acetyltransferase activity, hippocampal acetylcholine release, nicotinic and muscarinic (M1 and M2) receptor binding sites and acetylcholinesterase cell or terminal density showed no signs of alteration in either three-month or 9.5-month-old apolipoprotein E-deficient mice compared to controls. In contrast, long-term potentiation was found to be markedly reduced in these mice, but increases in the strength of stimulation induced the same level of long-term potentiation as that observed in controls. These alterations did not appear to be the consequence of modifications in the binding properties of glutamatergic receptors ( N-methyl- d-aspartate and [ RS]-α-amino-3-hydroxy-5-methylisoxazole propionic acid) but from defective regulation of the ( RS)-α-amino-3-hydroxy-5-methylisoxazole propionic acid receptor by phospholipase A2 activity. These results support the notion that apolipoprotein E plays a fundamental role in neuronal plasticity, which could in turn affect cognitive performance through imbalances in extra- and intracellular lipid homeostasis.

Reinforcement enhances hippocampal acetylcholine release in rats: an in vivo microdialysis study

Rats were trained to press a lever under a ‘Multiple FI-60s and Extinction’ schedule with food reinforcements. After learning the task, an in vivo microdialysis probe was inserted into the dentate gyrus and CA3 regions of the hippocampus, and the sequential changes in the dialysate acetylcholine (ACh) concentration were analyzed. In the session, two fixed-interval of reinforcement (FI) components (for 20 min) and two extinction (EXT) components (for 30 min) were alternated to examine the correlation between behavioral and neurochemical outcomes. The dialysate ACh concentration increased during the FI component and returned to the baseline during the EXT component of the schedule. Next, in order to dissociate the effect of discrimination from the effect of rewarding on the neurochemical changes in the hippocampus, we used a final TEST period (for 20 min) during which the actual schedule was extinction but the discriminative stimulus was on, i.e. the manifest condition of the test period was reinforcement. In the TEST period, the animals pressed the lever with almost the same frequency as during the FI component; however, the dialysate ACh concentration did not increase above the baseline concentration. These results suggest that ACh release in the rat hippocampus is associated with reinforcement but not with discrimination in operant conditioning.

Antisense oligodeoxynucleotides against the muscarinic m 2, but not m 4, receptor supports its role as autoreceptors in the rat hippocampus

Antisense oligodeoxynucleotides against muscarinic m 2 and m 4 receptors were used to investigate the role of these receptor subtypes as negative autoreceptors in the regulation of acetylcholine (ACh) release in the rat hippocampus. Following the continuous infusion of antisenses into the third ventricle (1 μg μl −1 h −1, 3 days), 3 H -AF-DX 384/muscarinic M 2-like binding was significantly decreased in the medial septum by the antisense against the m 2 receptor whereas M 2-like binding in the dorsal striatum was decreased by the antisense against the m 4 receptor. In contrast, 3 H -pirenzepine/muscarinic M 1-like binding was unaffected by either antisense treatment in any of the brain areas investigated. When perfused into the hippocampus via a dialysis probe, the purported muscarinic M 2 receptor antagonist AF-DX 384 (100 nM) increased hippocampal ACh release in freely moving rats. This effect of AF-DX 384 was significantly attenuated by the m 2, but not the m 4, receptor antisense treatment. Hippocampal choline acetyltransferase activity was not affected by either antisense treatments. Taken together, these results suggest that the molecularly defined muscarinic m 2 receptor regulates hippocampal ACh release by acting as a negative autoreceptor. In contrast, the molecularly defined m 4 receptor is unlikely to be directly involved in the negative regulation of ACh release in the rat hippocampus. Therefore, inhibiting muscarinic m 2 receptor function may be an alternative approach to regulate the release of ACh in neurodegenerative diseases associated with impaired cholinergic functions.

Serotonin and acetylcholine release response in the rat hippocampus during a spatial memory task

By using in vivo microdialysis we monitored the extracellular levels of acetylcholine and serotonin in the hippocampus of rats performing a spatial memory task. After rats were trained for 10 consecutive days to master a food-reinforced radial-arm maze task, they were implanted with a microdialysis probe in the dorsal hippocampus. On day 12, rats were tested in the maze and acetylcholine and serotonin outputs were monitored before the test, during the waiting phase and while performing the trials. In trained, food-rewarded rats, hippocampal acetylcholine levels increased during the waiting period (181±9% of baseline) and further increased during the radial-maze performance to 236±13% of baseline values, while serotonin levels did not change during the waiting period but increased to 142±3% during the maze performance. To discriminate whether the increase of acetylcholine and serotonin levels during the testing was associated with memory performance or with food consumption, we monitored hippocampal acetylcholine and serotonin release in rats that were trained, but not food rewarded, or in rats that were not trained, but rewarded only on the test day. In the trained, non-rewarded group, acetylcholine release increased during the waiting phase to 168±6%, but did not increase further during the task performance. In contrast, no change in serotonin release was observed in this group in any phase of the test. In rats which were not trained, but food rewarded, acetylcholine increased only during the maze period (150±5%). Serotonin increased gradually and become significant at the end of the trials. (130±3%). While both neurotransmitters could be implicated in feeding behaviour, only activation of cholinergic neurotransmission appears to be associated with memory function. Our results support the following hypotheses: (i) hippocampal acetylcholine could be involved in attentional and cognitive functions underlying motivational processes; (ii) serotonin could be implicated in non-cognitive processes (i.e. in the control of motor and feeding behaviour). Since serotonin and acetylcholine neurotransmission is simultaneously activated during the spatial memory task, this suggests that these neurotransmitter systems regulate behavioural and cognitive functions.

GABA receptors and benzodiazepine binding sites modulate hippocampal acetylcholine release in vivo

In the present study, the regulation of acetylcholine release from the ventral hippocampus by γ-aminobutyric acid (GABA) was investigated in vivo. GABA receptor agonists and antagonists were administered locally in the medial septum and the adjacent vertical limb of the diagonal band of Broca, or in the hippocampus by retrograde dialysis. Acetylcholine release was measured in the ventral hippocampus. In addition, the modulation of acetylcholine release via septal benzodiazepine binding sites was assessed by intraseptal administration of an agonists and an antagonist at the benzodiazepine binding site. Intraseptal administration of the GABAA receptor agonist muscimol and the GABAB receptor agonist baclofen, but not the agonist of the benzodiazepine binding site midazolam, decreased acetylcholine release in the hippocampus. The GABAA receptor antagonist bicuculline and the antagonist of the benzodiazepine binding site flumazenil, but not the GABAB receptor antagonist 3-N-(3,4,-dichlorobenzyl) aminopropyl-P-diethoxymethyphosphinic acid (CGP 52432) increased acetylcholine release in the hippocampus upon intraseptal administration. The same GABA receptor ligands were administered in the ventral hippocampus. CGP 52432 induced a small increase in acetylcholine release, whereas baclofen, muscimol and bicuculline did not affect local acetylcholine release. Thus, endogenous GABA causes tonic inhibition of acetylcholine release in the ventral hippocampus via septal GABAA receptors and, to a lesser extent, via GABAB receptors in the medial septum and hippocampus. The GABAergic inhibition in the medial septum is reduced by antagonists of the benzodiazepine binding site.

Inhibition of hippocampal acetylcholine release after acute and repeated Δ 9-tetrahydrocannabinol in rats

The effects of acute and repeated administration of Δ 9-tetrahydrocannabinol (Δ 9-THC), the psychoactive principle of marijuana, on acetylcholine release in the hippocampus was studied in freely moving rats by microdialysis. The acute intraperitoneal (i.p.) administration of Δ 9-THC at the doses of 2.5 and 5 mg/kg reduced acetylcholine release by about 25% and 45%, respectively. A dose of 7.5 mg/kg produced no further reduction. Δ 9-THC effects were antagonized by the cannabinoid CB 1 antagonist SR141716A at the i.p. dose of 1 mg/kg, per se ineffective in modifying acetylcholine concentrations. After a repeated exposure (twice daily for up to seven days) to Δ 9-THC (7.5 mg/kg, i.p.) or vehicle (0.3 ml/kg, i.p.), the inhibitory effect of Δ 9-THC (2.5 and 5 mg/kg, i.p) on acetylcholine release was not reduced. The results confirm previous observations that cannabinoids inhibit acetylcholine release through cannabinoid CB 1 receptors, and indicate that no tolerance to this effects develops after a repeated Δ 9-THC administration.

The specific dopamine uptake inhibitor GBR 12783 improves learning of inhibitory avoidance and increases hippocampal acetylcholine release

The specific dopamine uptake inhibitor, GBR 12783 was tested on the retention performance of a one-trial passive avoidance test. For a moderate electric shock intensity, GBR 12783 (10 mg/kg), injected before acquisition session, improved retention performance. Scopolamine (0.125–0.5 mg/kg) completely blocked the promnesic effect of GBR 12783. Moreover, GBR 12783 increased hippocampal acetylcholine release in vivo. These data suggest that the promnesic effect of GBR 12783 is mediated by an increase in the septo-hippocampal cholinergic transmission.

Restriction of environmental space attenuates locomotor activity and hippocampal acetylcholine release in male rats

We examined the effects of the restriction of environmental space on hippocampal acetylcholine release and spontaneous locomotor activity. Four days after the housing in a large or small cage, sampling for microdialysis study was begun. The locomotor activity counts exhibited significant daily changes in all rats in either the large or small cage. But, the mean locomotor activity counts in rats in the small cage was significantly less than that in the large cage. In contrast, the amount of acetylcholine collected per 20-min sample exhibited significant diurnal changes in all six rats in the large cage and in 5 of 6 rats in the small cage. The mean acetylcholine release in the rat in the small cage was significantly lower than that in the rat in the large cage during the dark phase, but not during the light phase. In addition, during the dark phase, hippocampal acetylcholine release was closely associated with spontaneous activity in all six rats in the large cage but not in 3 of 6 rats in the small cage. The present study suggests that the restriction of environmental space somehow interfere with the spontaneous locomotor activity and hippocampal acetylcholine release during the dark phase.

The effect of hypothalamic peptide YY on hippocampal acetylcholine release in vivo: implications for limbic function in binge-eating behavior

Central injection of peptide YY (PYY) in sated rats produces the most powerful stimulating effect of food intake known to date. The neural mechanisms by which PYY regulates appetite are not clear but may be important because abnormal levels of PYY have been implicated in the neurobiology of bulimia nervosa. Interactions between brain acetylcholine (ACh) and PYY had not been studied. Therefore, the present experiments were designed to explore the in vivo release of ACh from the hippocampus (HPC) of rats in response to hypothalamic infusion of PYY. Hippocampal ACh release was found to increase 400% in response to 10 μg PYY. In a separate experiment, blockade of the same area of the HPC with bilateral intracerebral injections of 3.5 μg scopolamine did not affect intake stimulated by intrahypothalamic injection of 4 μg PYY. Furthermore, a third experiment showed, for the first time, that PYY (2.5–10.0 μg) can elicit robust feeding when infused directly into the HPC. The significance of these findings to the activation of limbic functions such as memory, reinforcement, and obsessional processes that accompany human binge-eating syndromes is discussed.

Corticotropin-releasing factor administered centrally, but not peripherally, stimulates hippocampal acetylcholine release.

In addition to corticotropin-releasing factor's well-known role in mediating hormonal and behavioral responses to stress, this peptide also reportedly affects arousal and cognition, processes that classically have been associated with forebrain cholinergic systems. Corticotropin-releasing factor stimulation of cholinergic neurons might thus provide a mechanism for this peptide's cognitive effects. To examine this possibility, the present experiments characterize the effect of corticotropin-releasing factor on cholinergic neurotransmission, using in vivo microdialysis to measure hippocampal acetylcholine release. Corticotropin-releasing factor (0.5-5.0 microg/rat intracerebroventricularly) was found to increase dialysate concentrations of acetylcholine in a dose-dependent manner in comparison with a control injection, the ovine peptide having a greater effect than the same dose of the human/rat peptide. This effect was found to be centrally mediated, independent of the peripheral effects of an exogenous corticotropin-releasing factor injection; subcutaneous injections of the peptide increased plasma concentrations of corticosterone, the adrenal hormone ultimately secreted in the rat's stress response, to the same level as did the central injections, without affecting hippocampal acetylcholine release. These results demonstrate that corticotropin-releasing factor, acting centrally, regulates hippocampal cholinergic activity, and suggest that corticotropin-releasing factor/acetylcholine interactions may underlie some of the previously identified roles of these neurotransmitters in arousal, cognition, and stress.

Expression and presence of septal neurokinin-2 receptors controlling hippocampal acetylcholine release during sensory stimulation in rat.

We examined the expression and presence of NK2 receptors in the septal area of rat brain, and investigated their functional role in the regulation of the septohippocampal cholinergic system. Using reverse transcription-polymerase chain reaction (RT-PCR) analysis, we showed the presence of NK2 receptor mRNA expression in the septal area, and detected septal NK2 binding sites by using a fluorescent-tagged neurokinin A (NKA) derivative. In vivo microdialysis was employed to explore the functional role of NK2 receptors in the release of hippocampal acetylcholine evoked by tactile stimulation in freely moving rats. Two sessions of stroking of the neck and back of the rat for 30 min, at 90 min intervals, produced a marked and reproducible increase in hippocampal acetylcholine release. This effect was dose-dependently prevented by intraperitoneal administration of the two selective non-peptide tachykinin NK2 receptor antagonists SR144190 (0.03-0.3 mg/kg, i.p.) and SR48968 (0.3 and 1 mg/kg, i.p.), but not by the inactive enantiomer of SR48968 (SR48965, 1 mg/kg) nor by the two non-peptide NK1 receptor antagonists SR140333 (3 mg/kg, i.p.) and GR205171 (1 mg/kg, i.p.). Furthermore, the intraseptal application of SR144190 (10(-8) M) reduced the sensory response. Finally, intraseptal perfusion of neurokinin A (0.01-10 microM) in anaesthetized rats produced a concentration-dependent increase in hippocampal acetylcholine release. The response to neurokinin A (0.1 microM) was prevented by SR144190 (0.03-0.3 mg/kg, i.p.) and SR48968 (0.3-1 mg/kg, i.p.). In conclusion, this study provides direct evidence for the role of endogenous NKA/substance P, through the activation of NK2 receptors, in regulating the septohippocampal cholinergic function.

Coadministration of Galanin Antagonist M40 with a Muscarinic M1Agonist Improves Delayed Nonmatching to Position Choice Accuracy in Rats with Cholinergic Lesions

The neuropeptide galanin is overexpressed in the basal forebrain in Alzheimer's disease (AD). In rats, galanin inhibits evoked hippocampal acetylcholine release and impairs performance on several memory tasks, including delayed nonmatching to position (DNMTP). Galanin(1-13)-Pro2-(Ala-Leu)2-Ala-NH2 (M40), a peptidergic galanin receptor ligand, has been shown to block galanin-induced impairment on DNMTP in rats. M40 injected alone, however, does not improve DNMTP choice accuracy deficits in rats with selective cholinergic immunotoxic lesions of the basal forebrain. The present experiments used a strategy of combining M40 with an M1 cholinergic agonist in rats lesioned with the cholinergic immunotoxin 192IgG-saporin. Coadministration of intraventricular M40 with intraperitoneal 3-(3-S-n-pentyl-1,2,5-thiadiazol-4-yl)-1,2,5, 6-tetrahydro-1-methylpyridine (TZTP), an M1 agonist, improved choice accuracy significantly more than a threshold dose of TZTP alone. These results suggest that a galanin antagonist may enhance the efficacy of cholinergic treatments for the cognitive deficits of AD.

Stimulatory and inhibitory effects of ethanol on hippocampal acetylcholine release.

Using the microdialysis technique and sensitive HPLC procedures for the determination of acetylcholine (ACh) and ethanol, we investigated the release of ACh in rat hippocampus after acute ethanol administration. Systemic administration of ethanol (0.8 and 2.4 g/kg i.p.) led to peak ethanol concentrations of 21 and 42 mM in the hippocampus, respectively. The high dose caused a long-lasting inhibition of basal ACh release by up to 33%. Local infusion of scopolamine (1 microM) enhanced hippocampal ACh release up to eightfold in the presence of neostigmine (10 microM), and this stimulated release was also inhibited after systemic ethanol administration (by up to 45%). The low dose of ethanol (0.8 g/kg) led to a delayed stimulation of hippocampal ACh release. A stimulatory effect on ACh release was also observed when ethanol (50-100 mM) was infused directly into the hippocampus or into the septal area, i.e. to the origin of the cholinergic septohippocampal pathway; thus, the stimulatory effect may be mediated by a direct effect on cholinergic fibres. We conclude that ethanol exerts dual modulatory effects on the activity of the septohippocampal cholinergic fibres, depending on the dose and the site of administration. It is suggested that the inhibition of hippocampal ACh release by intoxicating doses of ethanol may contribute to the well-known cognitive and amnesic effects of ethanol intake.

Pharmacological characterization of nicotinic receptor-mediated acetylcholine release in rat brain—an in vivo microdialysis study

In vivo microdialysis was used to investigate nicotinic receptor-mediated acetylcholine release in the hippocampus, frontal cortex, and striatum of freely moving rats. Intraperitoneal administration of (−)-nicotine increased the release of acetylcholine in the hippocampus and frontal cortex but not in the striatum. (−)-Nicotine exhibited a bell-shaped dose–response relationship, and showed attenuation of response at the highest dose (5.0 mg/kg i.p.) in both the hippocampus and frontal cortex. In the hippocampus, (−)-nicotine (1.0 mg/kg i.p.)-induced increase of acetylcholine release was blocked by pretreatment with the centrally acting nicotinic receptor channel blocker, mecamylamine (1.0 mg/kg i.p.), but not by hexamethonium (5.0 mg/kg i.p.), suggesting that the effects of (−)-nicotine were mediated by the central nicotinic receptor. ( S)-3-methyl-5-(1-methyl-2-pyrrolidinyl)isoxazole (ABT-418, 1.0 and 5.0 mg/kg i.p.), reported to be a selective agonist for α4 β2 nicotinic receptor subunits, also enhanced the release of acetylcholine in the hippocampus, while 3-(2,4-dimethoxybenzlidene)-anabaseine (GTS-21, 1.0 and 5.0 mg/kg i.p.), which has high affinity for the α7 nicotinic receptor subunit, was without effect. The natural alkaloids isolated from plants, (−)-cytisine and (−)-lobeline, had little effect on acetylcholine release from the hippocampus. A competitive antagonist for α4 β2 subunits of the nicotinic receptor, dihydro- β-erythroidine, and a partial agonist for the β2 subunit-containing nicotinic receptor, (−)-cytisine, inhibited (−)-nicotine-induced increase of acetylcholine release from the hippocampus, whereas a selective antagonist for the α7 subunit, methyllycaconitine, and a partial agonist for the α3 subunit-containing nicotinic receptor, (−)-lobeline, did not. These results indicate that there are certain differences among brain regions in the response of nicotinic receptor-mediated acetylcholine release and that (−)-nicotine-induced acetylcholine release in the rat hippocampus may be attributed to activation of the α4 β2 nicotinic receptor subunits.