Yasushi Mizuki

Psychological stress increases dopamine turnover selectively in mesoprefrontal dopamine neurons of rats: reversal by diazepam.

The effects of psychological stress on catecholamine and indoleamine metabolism were examined in various brain regions of rats. Psychologically stressed rats were exposed to emotional responses of foot-shocked rats, but were themselves prevented from receiving foot-shock. Psychological stress for 30 min resulted in significant increases of both 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels in the medial prefrontal cortex (MPFC), but not in other dopamine (DA) terminal fields. The levels of noradrenaline (NA), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were unaffected in all brain regions examined after 30 min of psychological stress. A small but significant increase of DOPAC levels in the ventral tegmental area (VTA) was observed after a shorter (10 min) duration of stress. Moreover, an increase of DOPAC levels in the MPFC 30 min after psychological stress was attenuated by diazepam (5 mg/kg), and this attenuating effect was antagonized by Ro 15-1788 (15 mg/kg). These results suggest that mesoprefrontal DA neurons are selectively activated by psychological stress, and that the activation of the A10 cell body site (VTA) may precede that of the terminal field (MPFC). Moreover, diazepam was found to possess an inhibitory effect on the activation of mesoprefrontal DA neurons induced by psychological stress, and this effect may be partly mediated by benzodiazepine (BZD) receptors and implicated in the specific anxiolytic action of BZDs.

Differential responses to mental stress in high and low anxious normal humans assessed by frontal midline theta activity.

The distinct EEG theta rhythm from the frontal midline area observed during performance of mental tasks has been called Fm theta. In the present study, plasma catecholamine responses to mental stress were investigated using male students with (n = 12) and without (n = 12) Fm theta. The subjects were requested to complete the trait anxiety scale of STAI, and control blood samples were obtained. 65 min later, their EEGs were recorded during performance of an arithmetic addition task for 5 min. The state anxiety scores of STAI were obtained twice before and after the EEG recording. Blood samples were drawn three times during the state anxiety test and the EEG recording. The Fm theta appearance group showed low trait anxiety and a decrease of state anxiety after the mental task; however, the Fm theta non-appearance group exhibited high trait anxiety and no changes of state anxiety before and after the mental task. The concentrations of DA, HVA, NA and MHPG in the Fm theta appearance group showed lower levels at all times compared to those in the non-appearance group. In the Fm theta appearance group, an increase of DA turnover was observed by the addition of mental task. On the other hand, the Fm theta non-appearance group showed an increase of NA turnover.

Differential effects of noradrenergic drugs on anxiety and arousal in healthy volunteers with high and low anxiety

1. The appearance of frontal midline theta activity (Fm theta), the distinct EEG theta rhythm in the frontal midline area during performance of a mental task, indicates relief from anxiety in humans. 2. The authors investigated the effects of clonidine and yohimbine on anxiety and arousal in 24 male university students with (Fm theta group, n = 12) and without (non-Fm theta group, n = 12) Fm theta. Subjects received placebo, 0.15 mg clonidine and 15 mg yohimbine in a double-blind crossover design. 3. Blood samples were obtained, state-trait anxiety inventory (STAI) scores were determined, and EEGs were recorded before and during the performance of an arithmetic addition task. The test was repeated twice: before and 1 hr after drug administration. 4. Clonidine reduced the 3-methoxy-4-hydroxyphenylglycol (MHPG) concentration in both groups; yohimbine caused an increase in both groups. In the Fm theta group, clonidine reduced the appearance time of Fm theta and the number of task performance but did not alter the state anxiety scores; yohimbine had no effects on Fm theta or the state anxiety but increased the task performance. In the non-Fm theta group, clonidine increased the Fm theta amount and reduced the state anxiety score but did not affect task performance, while yohimbine reduced Fm theta but increased the state anxiety, the task performance and the number of errors. 5. These results suggest that changes in noradrenaline (NA) activity affect both anxiety and arousal levels in high-anxiety humans, but predominantly affect only the arousal level in low-anxiety humans.

Appearance of frontal midline theta rhythm and personality traits

Abstract: The distinct theta rhythm in the frontal midline area during a performance of mental tasks has been called Fmθ. However, Fmθ shows individual differences in its appearance. The relationship between the appearance of Fmθ and the subject personality was investigated in the present study. Forty male university students performed an arithmetic addition test for 5 min daily during three consecutive days and their EEGs were recorded during the test. They also completed the Maudsley Personality Inventory (MPI) and the Taylor Manifest Anxiety Scale (MAS) after the EEG recording on the last day. The subjects who obtained low scores on the anxiety scale of MAS, high scores on the extraversion scale of MPI and low scores on the neuroticism scale of MPI showed larger amounts of Fmθ. On the other hand, those subjects who showed high scores on the anxiety scale, low scores on the extraversion scale and high scores on the neuroticism scale showed smaller amounts of Fmθ or none. These results suggest that the appearance of Fmθ is closely related to the subject personality traits.

The mode of action of bromocriptine following pretreatment with reserpine and α-methyl-p-tyrosine in rats

The ability of bromocriptine (BRC), a selective dopamine D-2 receptor agonist, to induce yawning responses was studied in rats pretreated with reserpine and α-methyl-p-tyrosine (α-MPT). BRC (1–20 mg/kg IP) evoked yawning responses, which were pronounced at 2.5 mg/kg and characterized by the head moving downward. Higher doses of BRC (5–20 mg/kg) dose-dependently delayed the onset and peak time of yawning. A low dose of the selective D-1 dopamine receptor agonist SK&F38393 did not induce yawning but enhanced the BRC-induced response. Pretreatment with reserpine (1 and 5 mg/kg SC), α-MPT (100 and 300 mg/kg IP) and reserpine (1 mg/kg) plus α-MPT (100 mg/kg) was able to significantly reduce BRC-induced yawning. The inhibitory effects were prevented by a low dose of SK&F38393 (0.5 mg/kg IP). In particular, combined treatment with reserpine (5 mg/kg) and BRC (10 and 20 mg/kg) elicited upright fighting and jumping behaviors which were inhibited by haloperidol (1 mg/kg IP), a non-selective D-1 and D-2 receptor antagonist, SCH23390 (0.05 mg/kg SC), a selective D-1 receptor antagonist, or sulpiride (20 mg/kg IP), a potent D-2 receptor antagonist, and were potentiated by SK&F38393 (0.5 mg/kg). SCH23390 (0.05 mg/kg) decreased BRC-induced yawning and the apomorphine (low doses)-induced potentiation of BRC yawning, and prevented the apomorphine (high doses)-induced reduction of BRC yawning. SCH23390 also inhibited apomorphine-induced stereotypy and BRC-induced potentiation of apomorphine stereotypy. Furthermore, haloperidol (0.02 and 1.0 mg/kg IP), sulpiride (20 mg/kg IP) or scopolamine (0.5 mg/kg IP) inhibited BRC-induced yawning, but prazosin (1.0 and 3.0 mg/kg IP), an α-1 receptor antagonist, did not affect this behavior. These results suggest that BRC-induced yawning may be mediated via presynaptic dopaminergic neuron activity and that BRC, in addition to the stimulation of dopamine D-2 receptors, appears to require endogenous dopamine or receptor activation by another dopamine agonist (D-1 agonist) for the induction of yawning, stereotypy and upright fighting responses. The ability of dopamine agonists to induce these behaviors seems to depend apon the potency and ratio of D-2 versus D-1 receptor activity.

Dopaminergic and Cholinergic Interaction in Cataleptic Responses in Mice

The cataleptogenic effects of haloperidol, a dopamine D2 receptor antagonist; SCH23390, a D1 receptor antagonist; physostigmine, a cholinesterase inhibitor; and pilocarpine, a muscarinic M1 receptor agonist, were challenged by pretreatment of mice with SKF38393, a dopamine D1 receptor agonist; apomorphine, a dopamine D1/D2 receptor agonist (mainly D2 receptor); pirenzepine, a muscarinic M1 receptor antagonist; and scopolamine, a muscarinic M1/M2 receptor antagonist. The effect of physostigmine and pilocarpine on haloperidol and SCH23390 cataleptic responses was also examined. Each of the challenging agents blocked one or more of the cataleptogenic agents, but only scopolamine blocked all four. Pirenzepine blocked cataleptic responses induced by SCH23390 and pilocarpine, but not those by haloperidol and physostigmine. The results of this study suggest that the action of physostigmine (endogenous acetylcholine) on M2 receptors might be more potent than that on muscarinic M1 receptors. A further interesting observation was that the haloperidol-induced catalepsy was enhanced by physostigmine pretreatment, but not by pilocarpine pretreatment, whereas the SCH23390-induced catalepsy showed the opposite spectrum of enhancement by the two cholinergic agonists. We conclude that, although the four cataleptogenic agents act via the dopaminergic-cholinergic systems, their pharmacological differences may be due largely to the different receptor subtypes that are involved in the mediation of catalepsy produced by each agent. Thus, dopamine receptors not only influence the cholinergic muscarinic receptors, but muscarinic M1 and M2 receptors also might mediate dopamine D1 and D2 receptor responses, respectively. The results suggest that there are, at the least, relationships between muscarinic M1 receptors and dopaminergic D1 receptors, and between muscarinic M2 receptors and dopaminergic D2 receptors. Dopamine D1 and D2 receptors may interact in a synergistic fashion on dopaminergic systems, but act independently of each other in influencing other system such as cholinergic neurons.

Development of tolerance and reverse tolerance to haloperidol- and SCH23390-induced cataleptic effects during withdrawal periods after long-term treatment

The development of tolerance and reverse tolerance and reverse tolerance to the cataleptic effects of selective D1 antagonist, SCH23390, and the mainly D2 antagonist, haloperidol, was investigated in mice that had been chronically treated (7 or 30 days) with haloperidol (1 mg/kg SC), SCH23390 (0.5 mg/kg SC), or saline (5 ml/kg SC). In control animals, SCH23390 (0.1-1.0 mg/kg IP) and haloperidol (0.1-1.0 mg/kg IP) produced cataleptic responses in a dose-dependent manner, although the responses had different time course profiles. SCH23390 catalepsy had a rapid onset but a short duration, whereas haloperidol catalepsy had a slower onset and longer duration. This could be due to differences in lipid solubility of the drugs, or at least pertly to an action of the drugs on different neuronal pathways. The cataleptic effects of SCH23390 (0.3 mg/kg IP) and haloperidol (0.3 mg/kg IP) were significantly reduced in mice when given 24 h, but not 72 h, after the last dose of a 7 day-pretreatment course (short-term treatment) of SCH23390. However, after long-term treatment (30 days) with SCH23390, a challenge dose of SCH23390 exhibited reverse tolerance (i.e., increased catalepsy) when given 7-21 days, but not 1-3 days, after the last injection of the SCH23390 pretreatment course. In contrast, haloperidol catalepsy was not affected by long-term SCH23390 treatment. However, after the last dose of long-term haloperidol treatment both SCH23390 and haloperidol exhibited tolerance to their cataleptic effects at 1-3 days, a normal response at 7 days, and an exaggerated response (reverse tolerance) at 15-21 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of Dopamine Antagonists on Changes in Spontaneous EEG and Locomotor Activity in Ketamine-Treated Rats

We investigated the effects of dopamine antagonists on spontaneous cortical and hippocampal electroencephalographic (EEG) changes, and on hyperlocomotion in ketamine-treated rats. Ketamine (20-60 mg/kg IP) synchronized cortical EEG and desynchronized hippocampal EEG in a dose-dependent manner indicating that the drug induced a dissociation between the cortical and hippocampal EEG. These EEG changes were accompanied by an increase in spontaneous locomotor activity, which involved lack of focused direction, stereotypy, irritability and other abnormalities. Dopamine antagonists, such as haloperidol (0.3-1 mg/kg IP), and nemonapride (0.3-1 mg/kg IP), reversed the dissociation between the cortical and hippocampal EEG in ketamine (60 mg/kg IP)-treated rats. Ketamine-induced hyperlocomotion was also decreased by administration of haloperidol (0.3 and 1 mg/kg IP) or nemonapride (0.1-1 mg/kg IP). Thus, it was found that dopamine antagonists reversed the EEG alterations and behavioural changes in ketamine-treated rats.

A Physiological Marker for Assessing Anxiety Level in Humans: Frontal Midline Theta Activity

Abstract: The distinct theta rhythm in the frontal midline area during performance of mental tasks has been designated as Fmθ. Sixteen male university students who failed to show any appearance of Fmθ in 3 consecutive days were given diazepam 5 mg, amobarbital 80 mg, methylphenidate 15 mg and placebo, in a double‐blind, crossover design. Scores were made on the state anxiety scale of STAI; EEGs were recorded before and during performance of an arithmetic addition. The test was repeated twice: before and one hr after drug administration. Fmθ appeared following the drug administration even in those who had never shown the appearance of Fmθ, and the appearance time of Fmθ increased in the following order: diazepam > amobarbital > placebo > methylphenidate. The scores of STAI decreased in the same order. The speed of performed tasks was increased by methylphenidate and placebo, but decreased by amobarbital and diazepam. These results suggest that relief from anxiety might be reflected in the appearance of Fmθ and that Fmθ might be a useful tool to measure the anxiety level in humans.

The role of adenosinergic, GABAergic and benzodiazepine systems in hyperemotionality and ulcer formation in stressed rats

The effects of benzodiazepines, GABA and adenosine on distress-induced hyperemotionality and gastric lesion formation were investigated in rats. Hyperemotionality such as struggling, vocalization and defecation evoked immediately after immobilization stress were attenuated by diazepam, adenosine or adenosine plus diazepam. Conversely, pretreatment with these drugs produced rapid and potent exacerbation of gastric lesions observed after 12 h of stress. The potent adenosine A1-receptor agonist N6-cyclohexyl adenosine (CHA) markedly inhibited the distress-evoked hyperemotional behaviors and potentiated the ulceration. γ-Aminobutyric acid (GABA), muscimol, a GABA receptor agonist, and aminooxyacetic acid (AOAA), a GABA deaminase inhibitor, attenuated both stress-induced hyperemotionality and ulceration. The inhibitory effects of diazepam and GABA on hyperemotionality were reversed, respectively, by Ro15-1788, a benzodiazepine receptor antagonist, and bicuculline, a GABA receptor antagonist. The stimulatory effect of CHA on stress ulceration was potentiated by bicuculline but was not affected by Ro15-1788 or by picrotoxin, a chloride channel inhibitor. These results suggest that the mechanism involved in gastric lesion formation induced by immobilization stress may be different from that in hyperemotional behavior, and that the activation of GABAergic neurons may act as a central modulating factor in the hyperemotionality and ulceration induced by immobilization stress.