Masakatsu Takahashi

Facilitation of memory retrieval by pretest morphine mediated by μ but not δ and κ opioid receptors

Mice were trained to avoid electric shock (0.6 mA) in a step-through type passive avoidance learning task, retention being measured 24 h after the training trial. Morphine 10 mg/kg administered 30 min before the test trial (pretest) facilitated memory retrieval, and the effect was completely antagonized by 1 mg/kg naloxone, a selective μ-opioid receptor antagonist. On the other hand, pretest administration of 0.01–10 mg/kg DTLET, a selective δ-opioid receptor agonist, did not produce the same effect as morphine. Nor-binaltorphimine, a κ-opioid receptor antagonist, did not antagonize the effect of pretest morphine, at doses of 1 and 2 mg/kg. These results suggest that the facilitation of memory retrieval by pretest morphine is mediated through μ- but not δ- or κ-opioid receptors.

Retro-nociceptin methylester, a peptide with analgesic and memory-enhancing activity.

Retro-nociceptin methylester (retro-Noc-ME), which has an oppositely directed structure to that of nociceptin, showed weak affinity for nociceptin receptor and antagonized nociceptin-induced inhibition of contraction in a guinea pig ileum (GPI) assay. The peptide induced analgesia after intracerebroventricular (i.c.v.) administration at a dose of 100 nmol per mouse. Analgesia was not blocked by the opioid antagonist naloxone, which suggests that the analgesia is not mediated by opioid receptor. Furthermore, analgesia caused by retro-Noc-ME was not attenuated after repeated administration, that is, there was an absence of tolerance. The peptide improved learning ability after i.c.v. administration in a step-through experiment in mice.

The effect of ginseng extract on locomotor sensitization and conditioned place preference induced by methamphetamine and cocaine in mice

Repeated i.p. injections of 2 mg/kg methamphetamine (MA) or 20 mg/kg cocaine at 48-h intervals induced reverse tolerance to their ambulation-enhancing effects (behavioral sensitization). Furthermore, the reappearance of the sensitized state was observed at the time of readministration of MA or cocaine even after a 30-day discontinuation of drug administration. A concomitant injection of ginseng extract (GE), 200 mg/kg, i.p., suppressed the development of reverse tolerance and the reappearance of sensitization to MA and cocaine. Conditioned place preference to MA (1, 2, and 4 mg/kg, i.p.) and cocaine (1, 4, 10, and 20 mg/kg, i.p.), was completely blocked by GE, 200 mg/kg, i.p. combined treatment with MA of cocaine. Meanwhile, spontaneous motor activity and place preference were not affected by GE alone. These results provide evidence that GE may be useful clinically for the prevention of adverse actions of MA and cocaine.

Anti-analgesic and anti-amnesic effect of complement C3a

In the present study, we found that complement C3a exerted central effects after intracerebroventricular administration in mice. At doses of 3 and 10 pmol/mouse, the peptide showed an antagonistic effect on analgesia induced by morphine and U-50488H, known to be mu- and kappa-opioid receptor agonists, respectively. Moreover, complement C3a improved scopolamine- and ischemia-induced amnesia at a dose of 10 pmol/mouse. Anti-analgesia was not observed by C3a des-Arg at 10 pmol/mouse. The present findings suggest that complement C3a may act as a peptide with anti-opioid activity in the central nervous system.

FS stress induces long-lasting memory facilitation: Involvement of cholinergic pathways

We tested in vivo the hypothesis that foot-shock (FS) stress-induced prolongation of latencies in the one-trial step-through passive avoidance learning task in mice occurred through a long-term facilitation process. Whereas behavioral responses in control mice lasted for 24 h, decreasing progressively in the subsequent days, FS-stress exposure for 15 min before training (pretraining), immediately after training (posttraining), or 15 min before the test (pretest) resulted in a profound and sustained enhancement of test latencies that lasted for at least 96 h. These facilitating effects disappeared when FS exposure was delivered with a 2- or 3-h difference with respect to the training trial. Scopolamine (Scop) (1 mg/kg, intraperitoneally) 30 min before the training session caused impairment of test latencies in control and pretest stressed animals, but failed to affect both pre- and posttraining FS stress-induced enhancement. Our working hypothesis is that FS stress may increase the levels of acetylcholine in the presynaptic terminal or the firing rate of cholinergic input. Animals pretreated with FS stress daily for 1 or 4 days followed by the acute schedule described above showed no enhancements of test latencies. Pretraining Scop impaired test latencies in pre- and posttraining and pretest stressed animals, suggesting that unpredictability is a critical factor in activating behavioral long-term facilitation.

Dependency on the brain function of arginine vasopressin system of the development to and recovery from analgesic tolerance to morphine

Concomitant intracerebroventricular (i.c.v.) injection of anti-arginine vasopressin (AVP) antiserum dose-dependently suppressed the development of analgesic tolerance to daily morphine, 10 mg/kg, s.c., in mice. This suppressive effect of the antiserum was reduced by incubating the antiserum with AVP in vitro, before i.c.v. injection, suggesting that the antiserum inactivates brain AVP to result in the suppression of the development of tolerance in vivo. Similar to the antiserum, both AVP V1 and V2 antagonists given i.c.v., 10 ng and 20 ng/mouse, respectively, suppressed the development of morphine tolerance. Meanwhile, the administration of antiserum dose-dependently recovered morphine analgesia in morphine-tolerant mice and a complete recovery of analgesia was observed at the highest dose of antiserum following the second injection, and the effect of antiserum was maintained for 3 days after its withdrawal. Likewise, 10-100 ng/mouse of AVP V1 receptor antagonist given i.c.v. recovered morphine analgesia partially but significantly in a dose-dependent manner; however, AVP V2 receptor antagonist at the same doses partially recovered analgesic effect but the effect was neither significant nor dose-dependent. These findings suggest that the tolerance developed to morphine can be reversible when disturbing the function of brain AVP, but in addition to the different mechanisms of antiserum, V1 and V2 receptor antagonists, the V1 receptor-mediated mechanism may be more closely concerned in this phenomenon.

Achiral and chiral quantification of methamphetamine and amphetamine in human urine by semi-micro column high-performance liquid chromatography and fluorescence detection.

In this paper, miniaturized achiral and chiral high-performance liquid chromatographic procedures for the determination of methamphetamine and amphetamine in human urine are described. After a simple pretreatment of human urine (i.e., 10 microL of urine or diluted urine were acidified and dried-up under N2 at room temperature) and fluorescence derivatization with 4-(4,5-diphenyl-1H-imidazol-2-yl)-benzoyl chloride under mild conditions (pH 9.0, 10 min at room temperature), the derivatives were isocratically separated on a semi-micro ODS column with Tris-HCl buffer (0.1 M, pH 7.0): acetonitrile (45 + 55 v/v) at a flow rate of 0.2 mL/min or their enantiomers were separated on a semi-micro OD-RH column with sodium hexafluorophosphate (0.3 M aq.): acetonitrile (44 + 56 v/v) at a flow rate of 0.1 mL/min as the mobile phase. Wide-ranged calibration curves were obtained with detection limits for the achiral and chiral analyses in the atto and femtomol levels, respectively, per injected volume. Satisfactory within- and between-day reproducibility data were obtained with both the methods with the highest relative standard deviation being 9.6%. The methods were applied to the determination of methamphetamine and amphetamine in human urine samples and the concentrations determined by the two methods were well correlated (r = 0.994).

Demonstration of a human urinary trypsin inhibitor (urinastatin)-like substance in the murine brain

A human urinary trypsin inhibitor (urinastatin)-like immunoreactive substance with trypsin-inhibitory activities was found in the murine brain. Regional levels of this urinastatin (UT)-like substance in the brain were within 5 ng/mg protein and were expressed in a descending order as follows: cerebral cortex not equal to hippocampus > hypothalamus > mesencephalon not equal to corpus striatum >> medulla oblongata > cerebellum. Mechanical lesioning of the cerebral cortex or hippocampus with a needle induced a rapid and intense appearance of a UT-like immunoreactive substance in the neuronal cells of injured sites. Conditioned fear-stress induction incited a reversible increase in the level of UT-like immunoreactive substance in the hippocampus. These results suggest that the UT-like immunoreactive substance is produced by neurons in response to brain injury and fear-stress stimuli.

Attempts to Classify Dependence-Liable Drugs by Using a Simple Drug-Discrimination Test in Mice

1. In a simple discrimination test using a two-compartment shuttle box with mice, we examined the action properties of dependence-liable drugs. In mice trained to discriminate morphine from saline, neither methamphetamine (MAP) nor cocaine (COCA) was generalized to the discriminative stimulus effects of morphine. 2. Similarly, in mice trained to discriminate MAP from saline, COCA, which is known to have neuronal mechanisms in common with MAP, was generalized to the stimulus effects of MAP, but morphine was not. 3. Dihydroetorphine (DHE), which has receptor mechanisms in common with morphine, was generalized to the discriminative stimulus effects of morphine, whereas it was not generalized to the effects of MAP. Thus, the present discrimination test might be useful for the first screening of compounds with unknown neuronal mechanisms, particularly for classification into groups having separate neuropharmacological mechanisms in common.

Lack of the development of morphine tolerance in experimental amnesia: role of arginine vasopressin

The development of tolerance to morphine analgesia in amnesic model mice and the role of arginine vasopressin (AVP) in the underlying mechanism was examined. Hypoxia, brain ischemia, scopolamine and electroconvulsive shock (ECS) manipulation caused amnesia in the step-through type passive avoidance learning test performed at 24 h after the training trial. The amnesic state lasted for at least 3 days and recovered to naive control level on the 20th day after each manipulation. In all amnesic groups, radioimmunoassayable AVP content in hypothalamus was decreased, in particular, the reduction was significant in hypoxia and ischemic induced amnesic animals, then recovered to the control level by 20 days after each treatment. Daily morphine, 10 mg/kg, s.c. easily resulted in the development of tolerance to the analgesic effect in control animals; however, such treatment failed to develop tolerance in amnesic model animals, leaving the analgesic effect unchanged to the control levels. Daily pretreatment with i.c.v. AVP, dose-dependently reinstated the development of tolerance in amnesic model mice. When morphine injection was started from 20 days after the amnesia inducing treatment, tolerance developed as in a similar pattern as in control animals. Thus, amnesic model mice are deficient in brain AVP levels, and consequently, a certain level of AVP in the hypothalamus is required for maintaining the normal function such as the development of tolerance to morphine and the recovery from amnesia.