Markus Björklund

Cognitive Changes in Mice Following Moderate MPTP Exposure

We evaluated the cognitive effects of two moderate doses (30 mg/kg x 3 every 12 h and 20 mg/kg x 6 every 8 h, i.p.) of 1-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP) in mice. The dose of 30 mg/kg x 3 caused about 60% depletion of striatal dopamine but did not reduce the levels of its metabolites. Mice treated with MPTP did not differ from controls in their motor behavior in the open field. Mice treated with MPTP were comparable to controls in T-maze delayed alternation with fixed delays but were impaired when trials with mixed 20 s and 120 s delays were presented, indicative of a spatial working memory impairment. Dexmedetomidine at 10 microg/kg (s.c.) slightly improved delayed alternation performance in all groups but also slowed initiation of the motor response. Mice treated with MPTP at the dose of 30 mg/kg x 3 were less sensitive to this adverse effect of dexmedetomidine. The impairment in spatial working memory after MPTP exposure in mice parallels the findings in monkeys, but the deficit is much less severe.

Modulation of rat neocortical high-voltage spindle activity by 5-HT1/5-HT2 receptor subtype specific drugs

To investigate the role of serotonin (5-hydroxytryptamine; 5-HT) receptors in the modulation of rat thalamocortical oscillations, we studied the effects of 5-HT1/5-HT2 receptor subtype specific drugs on neocortical high-voltage spindle activity in adult male rats. A 5-HT1A receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) (0.03, 0.1, 0.3 and 1.0 mg/kg s.c.), had no effect on neocortical high-voltage spindle activity. Furthermore, a mixed 5-HT1/5-HT2 receptor antagonist, methysergide (1.0, 5.0 and 15.0 mg/kg i.p.), had no effect, whereas a non-specific mixed 5-HT1/5-HT2 receptor antagonist, methiothepin (0.2, 1.0 and 5.0 mg/kg i.p.), significantly increased neocortical high-voltage spindles. Of the 5-HT2 receptor antagonists, ritanserin (0.1, 1.0 and 5.0 mg/kg s.c.) had no effect, whereas ketanserin (1.0, 5.0 and 20.0 mg/kg s.c.) increased neocortical high-voltage spindles, but only at the highest dose used. A 5-HT2 receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.5, 1.0 and 2.0 mg/kg s.c.), at the two highest doses significantly decreased neocortical high-voltage spindle activity, and this effect was blocked by the 5-HT2 receptor antagonists, ketanserin (1.0, 5.0 and 20.0 mg/kg s.c.) and ritanserin (1.0 and 5.0 mg/kg s.c.), as well as by methiothepin (0.2, 1.0 and 5.0 mg/kg i.p.) and methysergide (1.0, 5.0 and 15.0 mg/kg i.p.). Furthermore, unilateral intrathalamic infusions, but not intrahippocampal control infusions, of DOI (10 and 50 micrograms/1.0 microliters/rat) decreased neocortical high-voltage spindle activity and systemic administration of ketanserin (20.0 mg/kg s.c.) completely blocked this effect. The present results suggest that (1) the serotonergic system modulates rat thalamocortical oscillations as measured by neocortical high-voltage spindle activity, (2) activation of 5-HT2 receptors, possibly located in the thalamus, with a specific 5-HT2 receptor agonist, DOI, causes a reduction in rat neocortical high-voltage spindle activity.

Lack of Relationship Between Thalamic Oscillations and Attention in Rats: Differential Modulation by an Alpha-2 Antagonist

A five-choice serial reaction time (5-CSRT) task was used to assess attention in rats. In this behavioral paradigm, the rats are required to spatially discriminate a short visual stimulus that will occur randomly in one of five locations while maintaining a sufficient activity level. The ability of a rat to maintain attention on the task can be measured by counting the choice accuracy (percent correct responses), whereas the probability of premature responses indicates the level of impulsivity. According to previous results [24], rats performing poorly in the task have a lower choice accuracy and make more premature responses than normally behaving individuals, i.e., a clear, inverse correlation was observed between choice accuracy and impulsiveness of rats. Methylphenidate, a psychostimulant that has been shown to alleviate the symptoms in attention deficit-hyperactivity disorder (ADHD), improved the choice accuracy of poor performing rats in this task [24]. The present results show that the correlation between choice accuracy and impulsivity exists also when the rats are tested using a reduced stimulus intensity or curtailed stimulus duration. The results of a pharmacological experiment suggested that atipamezole (30, 300, or 1000 micrograms/kg), a potent and specific alpha-2 antagonist that is known to increase the activity of monoaminergic systems in the brain, did not affect the percent correct responses in poor performers or in controls tested either at the baseline conditions or at a curtailed stimulus duration (which impaired their choice accuracy). At the doses of 300 and 1000 micrograms/kg, however, atipamezole slightly increased the probability of premature responses in all group of rats. The results of an electrophysiological study indicated that the poor choice accuracy or impulsiveness of rats is not related to the amount of cortically recorded spike-wave discharges/high voltage spindle (HVS) activity, which reflect thalamo-cortical oscillation. Atipamezole dose-dependently reduced the incidence and duration of HVSs. The present data, therefore, indicate that (a) alpha-2 antagonist treatment is not superior to methylphenidate treatment when investigated using acute administrations of the agents in poor performers of the 5-CSRT task, and (b) thalamic oscillations are not the reason for the attention deficit of rats in this model of ADHD. The relationship between choice accuracy and impulsivity is discussed.

Spatial working memory improvement by an α2-adrenoceptor agonist dexmedetomidine is not mediated through α2C-adrenoceptor

Abstract 1. Aged α2C-adrenoceptor knockout and wild type mice were used to investigate whether α2C-adrenoceptors are involved in mediating the beneficial effects of α2-adrenoceptor agonist, dexmedetomidine, on spatial working memory. 2. A win-stay task in the radial arm maze was used to dissociate the effects of dexmedetomidine on working vs. reference memory. In addition, the animals were tested in simple response habit learning in the T-maze. 3. Knockout mice made more working memory errors after the change of the baited arm in radial arm maze, but after training reached again as accurate level of performance as wild type controls. Dexmedetomidine 5 and 10 μg/kg alleviated the increase in spatial working memory errors after the change of the baited arm in knockout mice. Knockout and wild type mice performed equally well in T-maze, and dexmedetomidine had no effect on this simple response learning. 4. The present results indicate that α2-adrenoceptor agonists have a selective effect on spatial working memory not only in monkeys but also in mice. Further, this study confirms our earlier finding that the presence of α2C-adrenoceptors is not necessary for the spatial working memory enhancing effect of α2-adrenoceptor agonists.

An indirect cholinesterase inhibitor, metrifonate, increases neocortical EEG arousal in rats

WE investigated the ability of a cholinesterase inhibitor, metrifonate, to desynchronize cortical EEG activity. Metrifonate suppressed immobility-related high voltage spindling activity in young and aged rats at doses of 30 and 60 mg kg−1, p.o., and 10, 30 and 60 mg kg−1, p.o., respectively. The increase in EEG 1–20 Hz amplitude induced by scopolamine (0.2 mg kg−1, i.p.) was fully alleviated by metrifonate (30 and 100 mg kg−1, p.o.) and partially alleviated by a reference cholinesterase inhibitor, THA (3 and 6 mg kg−1, i.p.). Nucleus basalis (NB) lesions induced by quisqualic acid decreased frontal cortical choline acetyltransferase activity by 80% and increased cortical EEG slow waves. Metrifonate and THA did not reverse NB lesion-induced EEG abnormality. We conclude that metrifonate enhances cholinergic desynchronization of cortical EEG waves and that a severe defect of presynaptic NB cholinergic fibres limits the therapeutic effects of metrifonate.

Alpha2C-adrenoceptor mediated regulation of cortical EEG arousal

Alpha2-adrenergic drugs modulate cortical arousal and EEG. However, the role of individual alpha2-adrenoceptor (alpha(2)-AR) subtypes in these functions is not clear. We investigated the role of alpha(2C)-ARs in the modulation of baseline cortical EEG activity and EEG responses to the alpha(2)-AR selective agonist, dexmedetomidine (3-300 microg/kg, s.c.), and antagonist, atipamezole (3-1000 microg/kg, s.c.), by using alpha(2C)-AR knockout (KO) and wildtype (WT) mice. The overall amplitude (1-30 Hz) was not significantly altered in KO mice although the activity of theta band (4-8 Hz) was increased in these mice. The main finding was that dexmedetomidine (30-300 microg/kg) more effectively slowed and atipamezole (30-1000 microg/kg) less effectively increased cortical EEG arousal in KO mice compared to WT controls. Importantly, autoradiographical results showed no compensatory increase in other alpha(2)-AR subtypes in cortical, thalamic or other brain structures of KO mice. Furthermore, there were no differences between the genotypes in the levels of hippocampal choline acetyltransferase, monoamines or their metabolites. Altered baseline cortical EEG activity and EEG responses to alpha(2)-AR selective drugs in KO mice indicate that alpha(2C)-ARs are involved in regulation of cortical arousal. These results suggest that alpha(2C)-ARs may antagonize the sedative effect of alpha(2)-AR agonists mediated by activation of alpha(2A)-ARs.

Suppression of neocortical high-voltage spindles by nicotinic acetylcholine and 5-HT2 receptor stimulation.

To investigate the roles of the nicotinic acetylcholine receptor and the serotonin (5-hydroxytryptamine; 5-HT) subtype 2 receptor in the modulation of rat thalamocortical oscillations, the effects of systemic (s.c.) administration of nicotine, a nicotinic acetylcholine receptor agonist, and 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), a 5-HT2 receptor agonist, on neocortical high-voltage spindle activity occurring during quiet waking-immobility behavior in aged (28 months of age) and adult (7 months of age) rats were studied. Nicotine 0.1 and 0.3 mg/kg alleviated the age-related increase of neocortical high-voltage spindles, whereas in adult rats only nicotine 0.3 mg/kg was effective. DOI 0.3, 1.0 and 2.0 mg/kg suppressed high-voltage spindles in both aged and adult rats. In aged rats, a combination of subthreshold doses of nicotine (0.03 mg/kg) and DOI (0.1 mg/kg) decreased neocortical high-voltage spindles, whereas in adult rats two different subthreshold dose combinations (nicotine 0.03 or 0.1 mg/kg+DOI 0.1 mg/kg) had no effect. p-Chlorophenylalanine (400 mg/kg/day i.p. for 3 consecutive days) treatment decreased brain serotonin concentration (> 80% reduction), but did not affect high-voltage spindles. However, in both aged and adult rats, p-chlorophenylalanine treatment blocked the decrease in high-voltage spindle activity produced by DOI 0.3 mg/kg, though not the decrease produced by higher doses of DOI (1.0 and 2.0 mg/kg). It is important that, in adult rats, p-chlorophenylalanine treatment was able to abolish the decrease in high-voltage spindle activity seen after a relatively high dose of nicotine (0.3 mg/kg). The results suggest that nicotinic acetylcholine and 5-HT2 receptors may act in concert to suppress neocortical high-voltage spindling in rats, and that intact brain serotonergic systems may be important for some of the therapeutic effects of nicotine.

Activation of acetylcholine receptors and 5-HT2 receptors have additive effects in the suppression of neocortical high-voltage spindles in aged rats.

Abstract We investigated if activation of the muscarinic or nicotinic acetylcholine receptors and serotonin (5-hydroxytryptamine; 5-HT) subtype 2 receptors would have additive or synergistic effects on the suppression of thalamocortically generated rhythmic neocortical high-voltage spindles (HVSs) in aged rats. The 5-HT2 receptor antagonist, ketanserin, at a moderate dose (5 mg/kg) prevented the ability of a muscarinic acetylcholine receptor agonist, (oxotremorine 0.1 mg/kg), and a nicotinic acetylcholine receptor agonist (nicotine 0.1 mg/kg), to decrease HVSs. At a higher dose (20 mg/kg), ketanserin completely blocked the decrease in HVSs produced by moderate doses of muscarinic acetylcholine receptor agonists (pilocarpine 1 mg/kg and oxotremorine 0.1 mg/kg), and by a high dose of nicotine (0.3 mg/kg), though not that produced by high doses of pilocarpine (3 mg/kg) and oxotremorine (0.9 mg/kg). The ability of a 5-HT2 receptor agonist, (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.1–1.0 mg/kg), to suppress HVSs was non-significantly modulated by the nicotinic acetylcholine receptor antagonist, mecamylamine (1–15 mg/kg), and the muscarinic acetylcholine receptor antagonist, scopolamine (0.03–0.3 mg/kg). The effects of the drugs on behavioral activity could be separated from their effects on HVSs. The results suggest that activation of the muscarinic or nicotinic acetylcholine receptors plus 5-HT2 receptors has additive effects in the suppression of thalamocortical oscillations in aged rats.

The effects of cholinergic drugs on rat neocortical high-voltage spindles in ketanserin-treated rats.

To investigate the roles of the cholinergic system and 5-HT2 receptors in the modulation of thalamocortical oscillations, we studied the effects of systemic (s.c.) administration of anticholinesterases (physostigmine, tetrahydroaminoacridine) and muscarinic acetylcholine receptor agonists (pilocarpine, oxotremorine) on spontaneous thalamically generated rhythmic neocortical high-voltage spindles in adult rats pretreated with either saline or ketanserin, a 5-HT2 receptor antagonist. Ketanserin at 20.0 mg/kg increased the number of high-voltage spindles. In saline-treated rats, tetrahydroaminoacridine 3.0 and 9.0 mg/kg was able to decrease high-voltage spindles, whereas in ketanserin 20.0 mg/kg-treated rats only the highest dose of tetrahydroaminoacridine (9.0 mg/kg) decreased high-voltage spindles. Both doses of physostigmine, 0.12 and 0.36 mg/kg, decreased high-voltage spindles in both saline and ketanserin 20.0 mg/kg-treated rats. Lower doses of tetrahydroaminoacridine (1.0 mg/kg) and physostigmine (0.06 mg/kg) were ineffective in both saline- and ketanserin 20.0 mg/kg-treated rats. Pilocarpine 3.0 mg/kg and oxotremorine 0.1 and 0.9 mg/kg decreased high-voltage spindles in saline-treated rats. However, in rats treated with ketanserin 20.0 mg/kg, only the lower doses of pilocarpine (0.3 and 1.0 mg/kg) and oxotremorine (0.03 mg/kg) were able to decrease the high-voltage spindles. The results suggest that activation of the cholinergic system and activation of 5-HT2 receptors have additive effects in the suppression of thalamocortical oscillations and related neocortical high-voltage spindles in rats, thus maintaining effective information processing in thalamocortical networks.

Effects of combined nimodipine and metrifonate on rat cognition and cortical EEG

The present study investigated if short-term treatment with an L-type Ca2+-channel inhibitor, nimodipine, can stimulate cognitive functioning and cortical electroencephalograph (EEG) arousal, and potentiate the effect of a cholinesterase inhibitor, metrifonate. Pretraining administration of nimodipine (3, 10 and 30 mg/kg, p.o.) had no effect on water maze and passive avoidance behavior of young neurologically intact controls, or water maze and passive avoidance performance failure induced by scopolamine pretreatment (i.p.; 0.4 mg/kg during the water maze and 2.0 mg/kg during the passive avoidance study), medial septal lesioning, or aging. Furthermore, nimodipine (3, 10 and 30 mg/kg, p.o.) had no effect on the improvement by metrifonate (10 mg/kg, p.o.) of the water maze and passive avoidance failure induced by scopolamine pretreatment or medial septal lesioning, nor did it affect the potential of metrifonate (30 mg/kg. p.o.) to improve the water maze or passive avoidance behavior of aged rats. Finally, nimodipine (3, 10 and 30 mg/kg, p.o.) had no effect on spontaneously occurring thalamically generated neocortical high-voltage spindles or spectral EEG activity of young controls, nor did it alleviate the spectral EEG abnormality induced by scopolamine (0.2 mg/kg, i.p.) administration. Also, the combination of nimodipine 3 or 10 mg/kg and a subthreshold dose of metrifonate 10 mg/kg could not suppress high-voltage spindles or scopolamine treatment-induced spectral EEG activity abnormalities. According to the present results, short-term treatment with nimodipine does not stimulate cognitive functions or increase cortical EEG arousal, and does not block or potentiate the propensity of metrifonate to improve cognitive performance of rats.