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Emerging cholinergic mechanisms and ontogeny of response inhibition in the mouse.

Mice, 7, 11, 15, 19, and 85-115 (adult) days of age, served as subjects in experiments assessing effects of anticholinergics on the development of behavioral inhibition. The centrally active anticholinergic scopolamine produced a dose-dependent elevation in locomotor activity in 19-day-old and adult mice. Acquisition and retention of a step-off passive avoidance response (PAR) was initially studied in nondrugged subjects. Mice as young as 7 days of age learned and retained the PAR for 1 hr. Twenty-four-hour savings, however, were not observed until 19 days of age. Simple PAR performance deficits following scopolamine injection were first seen at 15 days of age. Mice in those age groups exhibiting 24-hr retention (19-day-olds and adults) were used to assess carry-over effects of scopolamine on retest. Only in the case of juveniles did scopolamine, injected prior to training, disrupt 24-hr retest performance. Since methylscopolamine, a peripherally active anticholinergic, had no effect on activity and PAR performance, it is assumed that scopolamine's effects were of central origin. The results suggest that behavioral suppression comes under cholinergic control during the second and third postnatal weeks but that cholinergic mechanisms may not mediate response inhibition uniformly throughout development.

Age Factors↗

Aging and cholinergic modulation of the transient magnetic 40-Hz auditory response.

Cholinergic blockade by scopolamine, a central muscarinic receptor antagonist, may produce transient memory impairment in healthy subjects, and it has been used as a neurochemical model of cognitive degeneration in aged individuals. To observe the muscarinic modulation of memory and cortical auditory processing, nine cognitively intact elderly subjects (59-80 years) were studied using neuropsychological tests and 122-channel magnetoencephalography (MEG) after an administration of scopolamine hydrobromide (0.3 mg, i.v.) or glycopyrrolate (0.2 mg, i.v.), a peripheral muscarinic antagonist. A double-blind randomized crossover design was used in two sessions separated by at least 1 week. Scopolamine, but not glycopyrrolate, produced a transient impairment of verbal memory performance in the elderly subjects. MEG indicated that the auditory-evoked 40-Hz magnetic response was significantly larger after scopolamine than after glycopyrrolate administration. Furthermore, reanalysis of our earlier results in younger subjects (20-31 years), basically supporting the present MEG findings, tentatively suggests that the scopolamine effects on the 40-Hz response may be slightly pronounced with aging. In sum, the transient magnetic 40-Hz auditory response may be useful in studies on brain cholinergic deficits in elderly subjects.

Aged↗

Effects of ZK 93426 on muscarinic and nicotinic antagonist or nucleus basalis lesioning-induced electrocortical slowing.

The present study investigated the effects of a benzodiazepine receptor antagonist, beta-carboline ZK 93426 (1, 3 and 10 mg/kg, IP), on scopolamine and nucleus basalis (NB) quisqualic acid lesion-induced neocortical electrocortical activity slowing in rats. Scopolamine induced a dose dependent increase in EEG spectral values and slow delta waves (0.3 < 0.9 = 2.7 mg/kg IP). ZK 93426 partially reversed EEG slowing induced by the smallest scopolamine dose (0.3 mg/kg), but had no effect on the EEG changes induced by higher doses. A combination of scopolamine at 0.3 mg/kg and mecamylamine (a centrally active nicotinic antagonist) at 10 mg/kg induced an EEG slowing that was not reversed by ZK 93426. NB lesions markedly decreased cortical choline acetyltransferase (ChAT) activity (-77%) and increased EEG slow waves. ZK 93426 had no effect on the NB lesion-induced slow wave activity increase. The present results support the idea that beta-carboline ZK 93426 may increase cortical cholinergic activity by disinhibiting the NB cholinergic neurons. However, if the activity of "NB to cortex" cholinergic system is greatly decreased by either a marked reduction in NB cell number (in NB-lesioned rats), a near complete cortical post-synaptic muscarinic receptor blockade (large scopolamine dose) or by a combination of nicotinic (decrease acetylcholine release) and muscarinic receptor blockade, the effects of beta-carboline ZK 93426 on EEG slowing may be negligible.

Animals↗

Combined treatment with a 5HT1A receptor agonist and a muscarinic acetylcholine receptor antagonist disrupts water maze navigation behavior.

The present study was designed to investigate the effects of combined treatment with a serotonin (5-HT)1A receptor agonist, 8-hydroxy-2-(dipropylamino)-tetralin (8-OH-DPAT), and a muscarinic acetylcholine receptor antagonist, scopolamine, on water maze (WM) navigation. Treatment with either 8-OH-DPAT or scopolamine before daily behavioral training disrupted spatial navigation at medium doses and cue navigation at high doses. Pretraining treatment with a combination of subthreshold doses of 8-OH-DPAT and scopolamine impaired WM spatial and cue navigation, but did not impair the WM performance if the drugs were injected post-training. In trained rats, combined injections of subthreshold doses of 8-OH-DPAT and scopolamine given pretraining did not impair the rats' ability to find the platform in a familiar or in a novel position. The combination of 8-OH-DPAT and scopolamine also disrupted WM navigation in rats with central 5-HT depletion. A combination of a peripheral muscarinic acetylcholine receptor antagonist and 8-OH-DPAT had no effect on WM navigation. These data suggest that combined treatment with drugs blocking muscarinic acetylcholine receptors and activating 5-HT1A receptors greatly impairs WM learning/performance, but does not impair spatial memory per se.

8-Hydroxy-2-(di-n-propylamino)tetralin↗

Effects of purine analogues on spontaneous alternation in mice.

The Y-maze was used to examine the effects of purines acting at A1 and A2 adenosine receptors upon spontaneous alternation, a model of working memory, in mice. In support of previous work, scopolamine produced a loss of spontaneous alternation behaviour to the 0.5 chance level. The A1 receptor selective agonist N6- cyclopentyladenosine (CPA) did not change spontaneous alternation behaviour alone, but it prevented the decrease of spontaneous alternation scores produced by scopolamine. The A1 receptor selective antagonist 1,3-dipropyl-8-cyclopentylxanthine (CPX) blocked the effect of CPA in combination with scopolamine but had no effect alone. The A2 receptor selective agonist (N6-[2-(3,5-dimethoxyphenyl)-2-(2- methylphenyl)ethyl] adenosine (DPMA), and the A2 receptor selective antagonist 3,7-dimethyl-1-propargylxanthine (DMPX) had no effect of alternation behaviour alone and did not modify the effect of scopolamine. The results indicate the ability of A1 but not A2 receptor activation to modify working memory deficits induced by scopolamine, but suggest that endogenous adenosine does not normally participate in working memory processes.

Adenosine↗

Arterial hypoxaemia following premedication in patients with coronary artery disease.

The effect of premedication with morphine and scopolamine, morphine alone, and scopolamine alone on arterial oxygen tension has been studied in patients with coronary artery disease undergoing bypass operations. Control arterial blood samples were obtained with the patients in the supine position breathing room air the day before operation. A second arterial blood sample was taken in the induction room with the patient breathing room air in the supine position. In the first group (scopolamine only) and second group (morphine only) cH+ (pH), PaCO2 and PaO2 did not change. In the third group (morphine and scopolamine), however, PaO2 decreased, while PaCO2 and cH+ increased (pH decreased). The changes in cH+ and PaCO2 were statistically significant, but of no clinical importance. The present study suggests that, when morphine and scopolamine premedication is used for patients with coronary artery disease, simultaneous administration of oxygen is important in helping to eliminate a possible cause of preoperative myocardial ischaemia.

Adult↗

Disruption of trace conditioning of the nictitating membrane response in rabbits by central cholinergic blockade.

Central muscarinic cholinergic involvement in classical conditioning of eyeblink responses was determined in trace and delay paradigms. Rabbits were trained on a trace procedure in which a 250-ms tone conditioned stimulus (CS) and a 100-ms air-puff unconditioned stimulus (UCS) were presented with a 500-ms trace interval. Each training session day consisted of ten tone alone, ten air-puff alone and 80 paired CS-UCS trials. Scopolamine hydrochloride at doses of 0.03 and 0.1 mg/0.5 ml per kg, s.c. dose-dependently disrupted acquisition of conditioned responses. Rabbits that were treated with scopolamine and failed to learn showed a gradual increase in conditioned responses during an additional training period with saline injections and no transfer from earlier training. Scopolamine methyl bromide, which does not appreciably cross the blood-brain barrier, showed no effects in the trace conditioning paradigm at a dose of 0.1 mg/kg, s.c., indicating central cholinergic blockade is responsible for the suppressive effect of scopolamine. Scopolamine hydrochloride at a dose of 0.1 mg/kg, s.c. did not block acquisition in the delay procedure with a 250-ms inter-stimulus interval, although the rate of acquisition was somewhat reduced by the drug. These data are the first to demonstrate that classical conditioning of the eyeblink response in the trace procedure is highly sensitive to central cholinergic deficits.

Animals↗

Involvement of cholinergic systems in the deficit of place learning in Morris water maze task induced by baclofen in rats.

Effects of oxotremorine on the deficit of place learning in the Morris water maze task induced by baclofen and scopolamine were examined to determine the involvement of brain cholinergic systems in the deficit of learning induced by baclofen. Rats were given 4 training trials per day with the submerged platform at a fixed location in the maze for 4 days. On day 4, rats were required to swim in the pool without the platform after the 4th training trial (probe test). Baclofen as well as scopolamine dose-dependently increased the escape latency in the training trials. In the probe test, baclofen as well as scopolamine dose-dependently reduced the duration in the quadrant where the platform had been originally located. Increased latency in the training trials and reduced duration in the probe test induced by scopolamine were dose-dependently attenuated by oxotremorine. Increased latency and reduced duration in the baclofen-treated rats were improved by oxotremorine as well as 2-hydroxysaclofen. Baclofen but not scopolamine induced motor incoordination in the rotarod test. Oxotremorine failed to improve motor incoordination induced by baclofen. These results suggest that cholinergic systems may be involved in the deficit of place learning induced by baclofen, and that the ameliorative effects of oxotremorine may not be due to improvement of motor incoordination.

Animals↗

Dopamine--acetylcholine "balance" in nucleus accumbens and corpus striatum and its effect on hypothalamic self-stimulation.

Three experiments investigated the suppression of hypothalamic self-stimulation in rats by neuroleptics and its restoration by centrally acting anticholinergic agents. Scopolamine (0.1--1.0 mg/kg i.p.) and benztropine (1.0--10.0 mg/kg i.p.) each enhanced self-stimulation when administered alone, and partially restored performance suppressed by spiroperidol (0.05--0.15 mg/kg i.p.). Benztropine strongly inhibits transmitter reuptake at DA synapses but scopolamine does not, thus inhibition of DA reuptake cannot fully account for the stimulant or antineuroleptic action of anticholinergic drugs. Neuroleptic and anticholinergic effects on self-stimulation rate were mutually subtractive, and statistical evidence of interaction was not obtained. Scopolamine was shown also to restore performance extinguished by discontinuation of the stimulating current. Smaller doses of scopolamine (50 nmol; 19 microgram) injected directly into the nucleus accumbens septi partially restored responding suppressed by spiroperidol, though similar doses of scopolamine injected bilaterally into the caudate-putamen were ineffective. These findings suggest that hypothalamic self-stimulation may be influenced by ACh-and DA-containing systems which exert independent effects on a third system controlling performance. These effects appear to reflect the level of arousal or motivation rather than the reinforcement process itself.

Acetylcholine↗

Effects of muscarinic receptor agonists and anticholinesterase drugs on high voltage spindles and slow waves.

The effects of muscarinic agonists (AF102B, pilocarpine, oxotremorine) and anticholinesterases (physostigmine, tetrahydroaminoacridine) were investigated on the incidence of thalamically generated rhythmic high voltage spindles and on scopolamine (0.2 mg/kg)-induced neocortical slow wave activity (i.e. increased sum amplitude value of the 1-20 Hz band in a quantitative electroencephalography (qEEG) analysis). AF102B and pilocarpine decreased high voltage spindles and scopolamine increased sum amplitude values at 3 and 9 mg/kg, but not at 1 mg/kg. Oxotremorine was less potent than AF102B or pilocarpine in suppressing high voltage spindles. Oxotremorine had no effect on the scopolamine-induced qEEG changes. Tetrahydroaminoacridine decreased high voltage spindles at 1, 3 and 9 mg/kg and slow waves at 9 mg/kg. Physostigmine decreased high voltage spindles and slow waves at 0.12 and 0.36 mg/kg. Based on the present results we propose that agonists possessing muscarinic M1 receptor activity are effective in decreasing high voltage spindles and scopolamine-induced slow wave activity, but agonists showing predominant muscarinic M2 receptor activity may be less effective in decreasing high voltage spindles and slow waves. Furthermore, tetrahydroaminoacridine decreased high voltage spindles at doses lower than those required to decrease scopolamine-induced slow waves. Physostigmine decreased high voltage spindles and slow waves over the same dose range. This result may indicate that non-cholinergic mechanisms are involved in the tetrahydroaminoacridine-induced decrease in high voltage spindles.

Alzheimer Disease↗

An inverted U-shaped curve for heptylphysostigmine on radial maze performance in rats: comparison with other cholinesterase inhibitors.

The potential of heptylphysostigmine tartrate (pyrrolo [2,3b] indol-5-ol, 3,3a,8,8a-hexahydro-1,3a,8-trimethylheptylcarbamate [ester, (3aS-cis)]) (MF201), a new second-generation cholinesterase inhibitor, to antagonize scopolamine-induced amnesia in rats was assessed in an 8-arm radial maze. Upon completing the training session, the rats were orally administered increasing doses of MF201 (2, 3, 4, 6 and 8 mg/kg) 60 min prior to a s.c. injection of scopolamine (0.25 mg/kg). 9-Amino-1,2,3,4-tetrahydroamino-acridine hydrochloride hydrate (tacrine) (0.25, 0.37, 0.5, 1 and 2 mg/kg), 1-benzil-4-[(5,6-dimethoxy-1-indanon)-2-yl]-methyl piperidine (E2020) (0.125, 0.18, 0.25 and 0.5 mg/kg) and physostigmine (0.15, 0.25, 0.5 and 1 mg/kg) were orally administered and rats were tested in the same task. As previously described, scopolamine induced an impairment in radial maze performance, measured in terms of total number of errors, total time taken to complete the task and the percentage of amnesic animals. The reversal of scopolamine-induced impairment was characterized by the presence of an inverted U-shaped dose-response curve. A significant antagonistic effect was achieved with a dose (mg/kg) of 0.25 for E2020, 0.5 for tacrine and physostigmine and 3, 4 and 6 for MF201, the latter manifesting a broader spectrum of activity (3-6 mg/kg). While the maximal active doses restored the scopolamine-induced modified pattern of arm entry, they were ineffective in reducing hypermotility, suggesting the drugs have a specific effect on cognitive function.

Amnesia↗

Concurrent blockade of beta-adrenergic and muscarinic receptors disrupts working memory but not reference memory in rats.

In order to clarify the interactions between monoaminergic and cholinergic systems in working and reference memory functions, the effects of administration of the alpha, beta-adrenergic, D1-, D2-dopaminergic or serotonergic receptor antagonist together with the muscarinic receptor antagonist scopolamine on this behavior were examined using a three-panel runway task. Both in working and reference memory tasks, the number of errors (attempts to pass through two incorrect panels of the three panel-gates at four choice points) was significantly increased by 0.32 mg/kg scopolamine, but not by the doses up to 0.18 mg/kg. The beta-adrenergic antagonist propranolol at 10 mg/kg had no effect on the number of working memory errors. Combined administration of 10 mg/kg propranolol and scopolamine at 0.1 and 0.18 mg/kg significantly increased the number of working memory errors. However, in a reference memory task, propranolol at 10 mg/kg did not affect the number of errors, whether administered alone or together with 0.1 mg/kg scopolamine. Other monoaminergic receptor antagonists, including the alpha-adrenergic antagonist phentolamine (3.2 and 10 mg/kg), D1-antagonist SCH23390 (0.032 and 0.056 mg/kg). D2-antagonist sulpiride (100 mg/kg) and serotonin antagonist cinanserin (10 and 32 mg/kg) had no significant effect on working or reference memory errors, whether they were administered independently or in combination with scopolamine at 0.1 mg/kg. These results suggest that beta-adrenergic/muscarinic interactions play an important role in mediating processes involved in working memory performance of rats.

Adrenergic beta-Antagonists↗

Detection of cholinergic mediation of behavior in 7-, 9-, and 12-day old rat pups.

A training procedure that permitted infant rats (7, 9, and 12 days old) to acquire a T maze discrimination to escape footshock (Experimental 1) was used to study the effects of the cholinergic antagonist, scopolamine hydrobromide (0, 0.2 mg/kg and 0.8 mg/kg) (Experiment 2). These results indicated that (1) Scopolamine had no effect on T maze choice behavior, unlike previous results for 15- and 23-day old rats, but (2) scopolamine did increase latency to choice in the discrimination task at all ages, an effect previously seen in 15-day old animals, but opposite that seen after scopolamine administration to 23-day old animals or adults. This "paradoxical" effect of scopolamine on response latency is present approximately two weeks prior to earlier estimates of the time course of maturation of the cholinergic system.

Age Factors↗

Brain localization of cholinergic influence on male sex behavior in rats: antagonists.

The muscarinic receptor antagonist scopolamine was microinjected into either the preoptic area or the lateral ventricle, preceding sexual behavior tests. In Experiment 1 unilateral ventricular injections of scopolamine delayed the initiation of copulation, while unilateral preoptic injections had no effect. In Experiment 2 bilateral injections into the preoptic area produced dose-related decreases in the percentages of animals intromitting and ejaculating. In Experiment 3 scopolamine, injected alone into the preoptic area, again decreased the percentages of animals mounting, intromitting, and ejaculating. The muscarinic agonist oxotremorine, injected alone into the preoptic area, decreased ejaculatory threshold (i.e., decreased the number of intromissions preceding ejaculation) as previously reported. Concurrent oxotremorine and scopolamine injections into the preoptic area were not different from vehicle; thus, scopolamine blocked oxotremorine's effect. These data suggest that some cholinergic activation of the preoptic area is critical for normal copulation, since bilateral blockade of muscarinic receptors there dramatically decreased the number of animals copulating. However, increased cholinergic activity there only reduced ejaculation threshold.

Animals↗

Vacuous jaw movements induced by acute reserpine and low-dose apomorphine: possible model of parkinsonian tremor.

Coadministration of the monoamine-depleting agent reserpine with a low dose of apomorphine has been shown to result in high levels of vacuous jaw movements in rats. Two experiments were conducted to study the pharmacologic and motoric characteristics of the vacuous jaw movements induced by 5.0 mg/kg reserpine plus 0.1 mg/kg apomorphine. The first experiment was undertaken to determine whether the vacuous jaw movements induced by reserpine plus apomorphine could be reduced by coadministration of the muscarinic antagonist scopolamine. Injections of scopolamine produced a dose-related decrease in vacuous jaw movements induced by reserpine plus apomorphine, with the two highest doses (0.5 and 1.0 mg/kg scopolamine) producing significant differences relative to the control group that received reserpine plus apomorphine. In the second experiment, a slow-motion videotape system was used to study the temporal characteristics of the vacuous jaw movements induced by reserpine and apomorphine, and to study the effects of 1.0 mg/kg scopolamine on these movements. Most of the vacuous jaw movements shown by rats treated with reserpine and apomorphine occurred in rapid bursts of jaw movement. Analysis of the interresponse times (i.e., time between each jaw movement) showed that most of the jaw movements had a local frequency in the range of 2.86-6.67 Hz. Cotreatment with scopolamine significantly affected several measures of jaw movements. Thus, the vacuous jaw movements induced by reserpine plus apomorphine can be reversed by anticholinergic treatment, and these movements tend to occur as periodic oscillations of the lower jaw with a frequency of 3-7 Hz. Vacuous jaw movements in rats share some characteristics with parkinsonian symptoms and may represent an animal model of parkinsonian tremor.

Animals↗

Cholinergic receptor antagonists impair formation of intermediate-term memory in the chick.

Several experiments examined the effects of cholinergic receptor antagonists on formation of memory in the chick. Scopolamine produced amnesia in chicks trained on a one-trial peck avoidance task in a dose-dependent manner. Pretraining injection of scopolamine produced amnesia that developed between 15 and 30 min after training, suggesting that scopolamine interferes with intermediate-term memory (ITM), previously described to be active during this time (Patterson, Alvarado, Warner, Bennett, & Rosenzweig, 1986). Pretraining injection of scopolamine or ouabain, an inhibitor of ATPase activity shown previously to inhibit formation of ITM, produced identical time courses of amnesia development, supporting the hypothesis that scopolamine interferes with ITM. Pirenzepine, an inhibitor of M1 muscarinic receptors, was effective in producing amnesia, whereas gallamine, an M2 receptor inhibitor, did not produce amnesia. These results suggest that M1, but not M2, receptors are involved in memory formation in the chick.

Animals↗

D2 dopaminergic regulation of striatal preproenkephalin mRNA levels is mediated at least in part through cholinergic interneurons.

The effect of administration of the muscarinic antagonist scopolamine on the increase in striatal preproenkephalin (PPE) mRNA following a 6-hydroxydopamine (6-OHDA) lesion or chronic D2 dopamine (DA) antagonist treatment was examined by dot-blot hybridization. Administration of scopolamine dose-dependently attenuated the 6-OHDA lesion-induced increase in striatal PPE mRNA. Administration of the D2 DA antagonist eticlopride to naive rats increased striatal PPE mRNA in a dose- and time-dependent fashion. Chronic coadministration of scopolamine attenuated the eticlopride-induced increase in striatal PPE mRNA. Chronic administration of scopolamine alone did not alter striatal PPE mRNA levels. In contrast, chronic administration of eticlopride, scopolamine or the two combined decreased striatal preprotachykinin (PPT) mRNA to the same extent, suggesting that there was no direct interaction between D2 dopaminergic and cholinergic mechanisms in the regulation of striatal PPT mRNA. These data indicate that DA differentially regulates striatal PPE and PPT mRNA and suggest that dopaminergic regulation of striatal PPE mRNA is mediated in part through D2 DA effects on striatal cholinergic neurons.

Acetylcholine↗

Muscarinic antagonists attenuate neurotensin-stimulated accumbens and striatal dopamine metabolism.

The effect of scopolamine and atropine upon the increase in extracellular 3,4-dihydroxyphenylacetic acid induced by central injection of neurotensin was examined in the nucleus accumbens and the striatum of anaesthetized rats using in vivo differential pulse voltammetry with carbon fibre electrodes. Scopolamine (1 and 3 mg/kg, i.p.) and atropine (20 micrograms, i.c.v.) did not alter the 3,4-dihydroxyphenylacetic acid level in the nucleus accumbens or the striatum, measured for 60 min after administration. Neurotensin (10 micrograms, i.c.v.) increased the 3,4-dihydroxyphenylacetic acid peak height in both regions. Pretreatment with scopolamine (1 mg/kg) 15 min before neurotensin injection blocked the increase in extracellular 3,4-dihydroxyphenylacetic acid in the striatum but not in the nucleus accumbens whilst scopolamine (3 mg/kg) partially attenuated the effect of neurotensin in the nucleus accumbens and blocked the increase in 3,4-dihydroxyphenylacetic acid in the striatum. Atropine partially attenuated the effect produced by neurotensin in the nucleus accumbens and blocked the increase in 3,4-dihydroxyphenylacetic acid induced by the peptide in the striatum. However, the increase in extracellular 3,4-dihydroxyphenylacetic acid induced by haloperidol (1 mg/kg, s.c.) was not altered by scopolamine (1 mg/kg) or atropine. Also, the increase in dopamine metabolism in the nucleus accumbens and the striatum after centrally injected haloperidol (10 micrograms, i.c.v.) was not altered by atropine (20 micrograms, i.c.v.). Together, the results demonstrate a functional interaction between muscarinic antagonists and neurotensin on in vivo dopamine metabolism in the nucleus accumbens and the striatum but with a greater effect in the latter region.

3,4-Dihydroxyphenylacetic Acid↗