Yoshiaki Miyamoto

Neurobehavioral alterations in mice with a targeted deletion of the tumor necrosis factor-α gene: implications for emotional behavior

Tumor necrosis factor-alpha (TNF-alpha) is emerging as an important modulator of the function of the central nervous system. In the present study, we investigated a role of endogenous TNF-alpha in cognitive and emotional function using mice with targeted deletions of the TNF-alpha gene. TNF-alpha-(-/-) mice showed normal diurnal rhythms of spontaneous locomotor activity and cognitive functions. Emotional behavior in the mutant mice, however, was significantly altered, which manifested in the performance in the open-field, elevated plus maze, and forced swimming tests. The altered performance in the elevated plus maze test was significantly alleviated by treatment with diazepam. Postmortem brain analysis of TNF-alpha-(-/-) mice revealed a significant increase in serotonin metabolism in the brain. These findings suggest a role for endogenous TNF-alpha in emotional behavior, which may possibly be related to alterations of serotonine metabolism.

Behavioural adaptations to addictive drugs in mice lacking the NMDA receptor ε1 subunit

N‐methyl‐d‐aspartate (NMDA) receptors, a subtype of glutamate receptors (GluRs) formed by assembly of the GluRζ subunit (called NR1 in rats) with any one of four GluRε subunits (GluRε1–4; NR2A–D), play an important role in excitatory neurotransmission, synaptic plasticity and brain development. Recent pharmacological studies have also indicated a role for NMDA receptors in drug addiction. In the present study, we investigated the behavioural adaptations to addictive drugs such as phencyclidine (PCP), methamphetamine (MAP) and morphine (MOR) in mice lacking the GluRε1 subunit of the NMDA receptor. GluRε1 mutant mice exhibited a malfunction of NMDA receptors, as evidenced by the reduction of [3H]MK‐801 binding in an autoradiographic receptor binding assay. GluRε1 mutant mice showed an attenuation of acute PCP‐ and MAP‐induced hyperlocomotion. The development of sensitization by repeated treatment with PCP and MAP at a low, but not high, dose was also suppressed. The development of MOR‐induced analgesic tolerance and naloxone‐precipitated MOR withdrawal symptoms were attenuated in GluRε1 mutant mice. In the place conditioning test, PCP‐induced place aversion in naive mice and place preference in PCP‐pretreated mice, as well as MOR‐induced place preference, were diminished whereas MAP‐induced place preference was not affected in GluRε1 mutant mice. These findings provide genetic evidence that GluRε1 subunit‐containing NMDA receptors are involved in certain aspects of drug addiction.

The role of tissue plasminogen activator in methamphetamine-related reward and sensitization

In the central nervous system, tissue plasminogen activator (tPA) plays a role in synaptic plasticity and remodeling. Our recent study has suggested that tPA participates in the rewarding effects of morphine by regulating dopamine release. In this study, we investigated the role of tPA in methamphetamine (METH)‐related reward and sensitization. Repeated METH treatment dose‐dependently induced tPA mRNA expression in the frontal cortex, nucleus accumbens, striatum and hippocampus, whereas single METH treatment did not affect tPA mRNA expression in these brain areas. The METH‐induced increase in tPA mRNA expression in the nucleus accumbens was completely inhibited by pre‐treatment with R(+)‐SCH23390 and raclopride, dopamine D1 and D2 receptor antagonists, respectively. In addition, repeated METH treatment increased tPA activity in the nucleus accumbens. There was no difference in METH‐induced hyperlocomotion between wild‐type and tPA‐deficient (tPA–/–) mice. On the other hand, METH‐induced conditioned place preference and behavioral sensitization after repeated METH treatment were significantly reduced in tPA–/– mice compared with wild‐type mice. The defect of behavioral sensitization in tPA–/– mice was reversed by microinjections of exogenous tPA into the nucleus accumbens. Our findings suggest that tPA is involved in the rewarding effects as well as the sensitization of the locomotor‐stimulating effect of METH.

Age-related expression of σ1 receptors and antidepressant efficacy of a selective agonist in the senescence-accelerated (SAM) mouse

The σ1 receptor is a unique intracellular receptor whose activation results in an efficient modulation of several neurotransmitter responses. Its role as a target for the rapid nongenomic effects of neuro(active)steroids and the age‐related diminutions in steroid levels suggested that targeting the σ1 receptor might allow alleviation of age‐related neuronal dysfunctions. We examined here the expression and behavioral efficacy of σ1 receptors in the senescence‐accelerated (SAM) mouse model. The σ1 receptor mRNA expression was measured by using comparative RT‐PCR in the olfactory bulb, hippocampus, hypothalamus, cortex, or cerebellum of senescence‐prone SAMP/8 and senescence‐resistant SAMR/1 control animals. No difference was observed between substrains in 6‐, 9‐, and 12‐month‐old (m.o.) mice. The σ1 protein expression was analyzed by using immunohistochemical techniques. Labeling was intense in the olfactory bulb, hippocampus, hypothalamus, and midbrain of both SAMR/1 and SAMP/8 mice, and the distribution appeared unchanged in 6‐, 9‐, and 12‐m.o. animals. The receptors in vivo availability was examined by using in vivo [3H](+)‐SKF‐10,047 binding. No age‐related difference was observed in the olfactory bulb, hippocampus, hypothalamus, cortex, cerebellum, and brainstem of 6‐ or 12‐m.o. SAMR/1 or SAMP/8 mice. The antidepressant efficacy of the selective agonist igmesine was examined in the forced‐swimming test. The compound decreased significantly the immobility duration at 60 mg/kg in 6‐ and 12‐m.o. SAMR/1 and in 6‐m.o. SAMP/8 mice. In 12‐m.o. SAMP/8 mice, the drug efficacy was facilitated; a significant effect was measured at 30 mg/kg. Decreased neurosteroid levels, particularly of progesterone, were seen in 12‐m.o. SAMP/8 mice that might explain the enhanced efficacy of igmesine. Preserved σ1 receptor expression and enhanced behavioral efficacy of σ1 agonists were measured in SAM animals, confirming the therapeutic opportunies for selective ligands against age‐related mood disorders.

Phencyclidine-induced discriminative stimulus is mediated via phencyclidine binding sites on the N-methyl-D-aspartate receptor-ion channel complex, not via sigma1 receptors

The effects of several N-methyl-D-aspartate (NMDA) receptor- and sigma receptor-related compounds on the discriminative stimulus effects of phencyclidine (PCP) were examined in rats trained to discriminate PCP (1.5 mg/kg, i.p.) from saline under a two-lever fixed ratio 20 schedule of food reinforcement. PCP produced a dose-dependent increase in PCP-appropriate responding. A non-competitive NMDA receptor antagonist, dizocilpine (0.2 mg/kg, i.p.) and a putative sigma(1) receptor agonist, (+)-SKF-10047 (10 mg/kg, i.p.) fully substituted for PCP in every rat tested. Neither a competitive NMDA receptor antagonist, CGS-19755 (0.1-3 mg/kg, i.p.), sigma(1) receptor agonist, (+)-pentazocine (10-30 mg/kg, i.p.) nor dextromethorphan (10-20 mg/kg, i.p.) produced PCP-like discriminative stimulus effects. The discriminative stimulus effects of PCP (1.5 mg/kg, i.p.), dizocilpine (0.2 mg/kg, i.p.) and (+)-SKF-10047 (10 mg/kg, i.p.) were significantly attenuated by CGS-19755 (1 mg/kg, i.p.), but not by sigma(1) receptor antagonist BMY-14802 (10 mg/kg, i.p.) and NE-100 (5 mg/kg, i.p.). These results suggest that the discriminative stimulus effects of PCP are predominantly mediated via PCP binding sites on the NMDA receptor-ion channel complex, not via sigma(1) receptors. In addition, the PCP-like discriminative stimulus effects of (+)-SKF-10047 were demonstrated to be mediated via PCP binding sites.

Effects of single and repeated administration of methamphetamine or morphine on neuroglycan C gene expression in the rat brain.

The rearrangement of neural networks associated with the behavioural sensitization and tolerance induced by psychostimulants is poorly understood. We have investigated the effects of repeated administration of methamphetamine (chronic MAP), which induces behavioural sensitization, or morphine (chronic morphine), which induces tolerance to its antinociceptive effect, on the mRNA levels of neural network-related genes in the rat brain. A gene of special interest was that for neuroglycan C (NGC), a neural tissue-specific transmembrane chondroitin sulphate proteoglycan. Single MAP (acute MAP) administration significantly decreased NGC mRNA levels in the frontal cortex, ventral tegmental area (VTA), and amygdala compared to vehicle-treated groups. Repeated MAP (chronic MAP) administration significantly increased NGC mRNA levels in the frontal cortex, nucleus accumbens (NAc), striatum, hippocampus, VTA, and amygdala compared to acute MAP treatment. Single morphine (acute morphine) administration significantly increased NGC mRNA levels in the NAc, striatum, hippocampus, VTA, and amygdala compared to vehicle-treated groups. Chronic morphine administration significantly decreased NGC mRNA levels in the NAc, striatum, VTA, and amygdala compared to acute treatment. In addition, the NGC protein level in the NAc was increased after chronic MAP and acute morphine treatment. Dopamine and opioid receptor antagonists attenuated the effect of MAP and morphine respectively on NGC mRNA levels. These results suggest that the sensitization to MAP is associated with up-regulation of NGC gene expression, while the tolerance to the morphine-induced analgesic effect is associated with the down-regulation of NGC gene expression.

Involvement of nitric oxide in phencyclidine-induced place aversion and preference in mice

The present study investigated the involvement of nitric oxide (NO) in phencyclidine (PCP)-induced place aversion and preference in the place conditioning paradigm. PCP-induced place aversion in naive mice was dose-dependently attenuated by administration of N(G)-nitro-L-arginine methyl ester (L-NAME), a NO synthase (NOS) inhibitor, during the conditioning. The NOS activity and dopamine (DA) turnover in the hippocampus in mice showing PCP-induced place aversion were decreased, such changes being restored by administration of L-NAME during the conditioning. On the other hand, PCP-induced place preference in mice pretreated with PCP for 28 days was not attenuated by administration of L-NAME during the conditioning. Although NOS activity was not changed, the DA turnover in the cerebral cortex was increased in mice showing PCP-induced place preference. In mice pretreated with L-NAME and PCP for 28 days before the place conditioning paradigm, PCP neither induced place preference, nor changed the NOS activity or DA turnover. These results suggest that NO is involved in the acquisition of PCP-induced aversive effects, and in the development of PCP-induced preferred effects. Further, the functional change of the DAergic neuronal system mediated by NO in the hippocampus and cerebral cortex may be necessary for the expression of aversive effects and development of preferred effects, respectively, induced by PCP.

Involvement of the serotonergic neuronal system in phencyclidine-induced place aversion in rats

The possible involvement of the serotonergic neuronal system in aversive motivation produced by phencyclidine [1-(1-phenylcyclohexyl)piperidine; PCP] was investigated using a place-conditioning paradigm in rats. PCP (4 mg/kg, i.p.) produced place aversion in this task as reported previously (Kitaichi K, Noda Y, Hasegawa T, Furukawa H, Nabeshima T. Acute phencyclidine induces aversion, but repeated phencyclidine induces preference in the place conditioning test in rats. Eur J Pharmacol 1996;318:7-9). The blockade of serotonin2A (5-HT2A) receptors using the antagonist ritanserin (3 and 10 mg/kg, p.o.) significantly attenuated this aversive property of PCP whereas lesions of serotonergic neurons using 5,7-dihydroxytryptamine (5,7-DHT, 100 microg/animal, i.c.v.) failed to affect it. Repeated PCP treatment (10 mg/kg, i.p. for 14 days), which is enough to diminish the stereotyped 5-HT2A receptor-mediated head-twitch behavior, also decreased the place aversion. These results suggest that the serotonergic neuronal system, specifically the 5-HT2A receptor, may play a critical role in producing PCP-induced place aversion.