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Antihistamines are widely used to treat allergy symptoms. First-generation antihistamines have adverse effects on the central nervous system (CNS), such as hypnotic and amnesic effects, whereas second-generation antihistamines have poor brain penetration, and therefore, have fewer CNS-related adverse effects. Memory consists of several phases, including acquisition, consolidation, expression, and extinction. It remains unclear whether these phases are affected by antihistamines. We investigated the effects of diphenhydramine, a first-generation antihistamine, and levocetirizine and olopatadine, second-generation antihistamines, on memory phases. Mice were subjected to fear conditioning on day 1 and tested on day 2. Antihistamines were administered before conditioning, immediately after conditioning, or before the test session. Diphenhydramine (30 mg/kg) decreased freezing time when administered immediately after conditioning or before the test session. These effects were not attributable to a change in locomotor activity. Levocetirizine (0.1, 1, 10 mg/kg) and olopatadine (1, 10, 20 mg/kg) had no effects on conditioned fear. We also examined the effect of diphenhydramine and levocetirizine on the expression of an activity-dependent gene associated with the test session. Diphenhydramine, but not levocetirizine, increased Arc transcription in the central nucleus of the amygdala. These data indicate that diphenhydramine, but not levocetirizine or olopatadine, impairs the consolidation and expression of conditioned fear.

Vagus nerve stimulation (VNS) is an approved treatment for epilepsy and depression and has cognition-enhancing effects in patients with Alzheimer's disease. The hippocampus is widely recognized to be related to epilepsy, depression, and Alzheimer's disease. One possible mechanism of VNS involves its effect on the hippocampus; i.e. it increases the release of noradrenaline in the hippocampus. However, the effect of VNS on synaptic transmission in the hippocampus is unknown. To determine whether VNS modulates neurotransmission in the hippocampus, we examined the effects of VNS on perforant path (PP)-CA3 synaptic transmission electrophysiologically in anaesthetized rats. VNS induces a persistent enhancement of PP-CA3 field excitatory post-synaptic potentials (fEPSPs). Arc, an immediate early gene, was used to identify active brain regions after VNS. The locus coeruleus (LC), which contains the perikarya of noradrenergic projections, harboured more Arc-positive cells, as measured by in-situ hybridization, after 10-min VNS. In addition, electrical lesions of LC neurons or intraventricular administration of the β-adrenergic receptor antagonist timolol prevented the enhancement of PP-CA3 responses by VNS. In conclusion, the protracted increase in PP-CA3 synaptic transmission that is induced by VNS entails activation of the LC and β-adrenergic receptors. Our novel findings suggest that information from the periphery modulates synaptic transmission in the CA3 region of the hippocampus.

The dentate gyrus (DG), a part of the hippocampal formation, is a candidate target of antidepressants and may play a role in the development of depressive syndrome; however, there is no direct neurobiological evidence supporting this theory. Here, we examined whether DG integrity is necessary for the behavioural effects of acute or chronic antidepressant treatment. Microinjection of colchicine into DG severely damaged the granule cells, as confirmed by morphological, electrophysiological, and behavioural analyses. Acute treatment with desipramine and fluoxetine decreased the immobility of saline-treated rats in the forced swimming test, whereas this decrease was inhibited in colchicine-treated rats. Chronic treatment with desipramine and fluoxetine also decreased the immobility of saline-treated rats; however, the extensive DG damage induced by colchicine had no effect on this decrease. In the novelty-suppressed feeding test, chronic treatment with desipramine and fluoxetine decreased the latency to feed in saline-treated rats while, once again, the extensive DG damage caused by colchicine had no effect on this decrease. Thus, we concluded that DG integrity was required for the behavioural effects of acute but not chronic antidepressant treatment; this disparity was not due to the time interval between surgery and behavioural tests. These findings indicate that treatment duration determines the influence of DG integrity on antidepressant effects.

熱性けいれんは乳幼児に頻発するけいれん発作である．この有熱発作には遺伝素因や，発熱および未熟な脳といった各種因子が関与することが示されている．熱性けいれんには単純型と複雑型が存在し，前者は良性とされている．一方で，後者に関しては，将来的に側頭葉てんかんの発症へ関与する可能性が示唆されている．内側側頭葉てんかん患者の海馬では，神経細胞死や，神経細胞形態の異常に基づく異所性神経回路が確認されており，これらは神経細胞群の同期した過剰発射を誘発することでてんかん焦点を形成する．従って，熱性けいれんが細胞・分子レベルで海馬に与える影響および，これがてんかん原性の獲得に関与する可能性を追及することは，てんかんの発症メカニズムを解明する上で重要となる．しかし，ヒトの検体は，主に病状が進行し様々な薬物治療履歴がある成人患者由来であるため，これのみを使用した研究では，上述の目的のためには限界があった．この問題を解決するために，優れたモデル動物を利用して主に以下の4点が精力的に研究されてきた．即ち，(1)高熱状態が熱性けいれんを誘起するメカニズム，(2)熱性けいれんが与える神経解剖学的変化，(3)熱性けいれんが与える神経生理学的変化，そして(4)熱性けいれんが与える記憶・学習に対する影響である．本総説では以上に関して現在までに集積された知見を評価しながら議論する．

Febrile seizures (FS) are the most common type of convulsive events in infancy and childhood. Genetic and environmental elements have been suggested to contribute to FS. FS can be divided into simple and complex types, the former being benign, whereas it is controversial whether complex FS have an association with the development of temporal lobe epilepsy (TLE) in later life. In the hippocampus of TLE patients, several structural and functional alterations take place that render the region an epileptic foci. Thus, it is important to clarify the cellular and molecular changes in the hippocampus after FS and to determine whether they are epileptogenic. To achieve this goal, human studies are too limited because the sample tissues are only available from adult patients in the advanced and drug-resistant stages of the disease, masking the underlying etiology. These facts have inspired researchers to take advantage of well-established animal models of FS to answer the following questions: 1) How does hyperthermia induce FS? 2) Do FS induce neuroanatomical changes? 3) Do FS induce neurophysiological changes? 4) Do FS affect the behavior in later life? Here we introduce and discuss accumulating reports to answer these questions. Keywords:: childhood seizure, epilepsy, hippocampus, dentate gyrus, granule cell, neurodisease

We describe the effect of high pressure and high temperature on neuronal activity. Increased intracranial pressure is generally a pathological sign observed in intracerebral hemorrhage, brain edema, and brain tumor, yet little is known about how the hyperbaric pressure per se affects neuronal activity. Using a pressure/temperature-changeable perfusion chamber, we carried out functional multineuron calcium imaging to record spontaneous spiking activity simultaneously from about 100 neurons in hippocampal slice cultures. High-pressure conditions (up to 100 mmHg) did not alter the network excitability, whereas high-temperature conditions (up to 40°C) increased synchronized network activity. Thus, neurons are sensitive to feverish conditions, but the acute hyperbaric circumstance itself is unlikely to exert a detrimental effect on neuronal function. Keywords:: pressure, calcium imaging, neuronal network

Although ethanol has been shown to impair acquisition of memory, its effect on consolidated memories is not clear. Recent reports revealed that memory retrieval converted consolidated memory into a labile state and initiated the reconsolidation process. In the present study, we have demonstrated the effect of ethanol on reactivated fear memory. We used contextual fear conditioning where rats were conditioned with mild footshock, re-exposed to the training context for 2 min, immediately injected with ethanol or saline, and finally tested 48 h after re-exposure. Ethanol-treated groups demonstrated longer freezing and the effect lasted for 2 weeks. Reactivation is necessary for this effect. Injection of ethanol itself did not induce a fearful response. Reactivated and ethanol-treated rats exhibited longer freezing than non-reactivated controls, suggesting that ethanol does not inhibit the memory decline but facilitates the fear memory. Two minute re-exposures induced no or little extinction. The effect of ethanol was specific for 2-min reactivation, which induces reconsolidation. Moreover, we found that picrotoxin inhibited the memory enhancement that was produced by ethanol administered just after the reactivation. These studies demonstrate that ethanol enhances reactivated contextual fear memories via activation of GABAA receptors. Neuropsychopharmacology (2008) 33, 2912–2921; doi:10.1038/npp.2008.13; published online 20 February 2008

Oseltamivir, a widely used anti-influenza drug, inhibits virus neuraminidase. A mammalian homologue of this enzyme is expressed in the brain, yet the effect of oseltamivir on central neurons is largely unknown. Patch-clamp recordings ex vivo revealed that oseltamivir enhanced spike synchronization between hippocampal CA3 pyramidal cells. Time-lapse multineuron calcium imaging revealed that oseltamivir and its active metabolite evoked synchronized population bursts that recruited virtually all neurons in the network. This unique, so-far-unknown, event was attenuated by muscarinic receptor antagonist. Thus, oseltamivir is a useful tool for investigating a new aspect of neural circuit operation.Supplementary Fig. and movie: available only at http://dx.doi.org/10.1254/jphs.SC0070467 Keywords:: oseltamivir, functional multineuron calcium imaging, network excitability

Colchicine, a potent microtubule-depolymerizing agent, is well known to selectively kill dentate granule cells in the hippocampal formation in vivo. Using organotypic cultures of rat entorhino-hippocampal slices, we confirmed that in vitro exposure to 1 μM and 10 μM of colchicine reproduced a specific degeneration of the granule cells after 24 h. Similar results were obtained with other types of microtubule-disrupting agents, i.e., nocodazole, vinblastine, and Taxol. Interestingly, the actin-depolymerizing agents cytochalasin D and latrunculin A also elicited selective neurotoxicity in the dentate gyrus without affecting survival of hippocampal pyramidal cells. The selective pattern of degeneration was observable 24 h after a brief treatment with the toxins as short as 5 min, but this delayed neuronal death was unlikely to be a result of excitotoxicity because it was virtually unaffected by glutamate receptor antagonists, tetrodotoxin, or extracellular Ca 2+ -free conditions. The damaged tissues contained a large number of TUNEL-positive neurons and exhibited an increased level in caspase-3-like activity, suggesting that cytoskeleton disruption triggers an apoptosis-like process in dentate granule cells. Thus, this study may provide a basis for understanding the distinctive mechanism that supports granule cell survival.

Although it is well known that the hippocampal CA1 subfield is highly vulnerable to ischemic injury, cellular mechanisms leading to this neuronal degeneration are not fully understood. Using organotypic cultures of rat hippocampal slices, we determined whether phospholipase A2 (PLA2) is activated in response to ischemic conditions (OGD; oxygen and glucose deprivation). The PLA2 activity in the pyramidal cell layer increased immediately following a 35-min exposure to OGD, which was likely to be mediated by selective activation of cytosolic Ca 2+ -dependent PLA2 subtype (cPLA2). This enhancement lasted for at least 24 h. Interestingly, no apparent increase was detected in the dentate gyrus. Twenty-four hours after the OGD exposure, neuronal death was detected mainly in the CA1 region of hippocampal slices. To examine whether the PLA2 activation is causally or protectively involved in the ischemic injury, we investigated the effect of pharmacological blockade of PLA2 on the OGD-induced neuronal death. The PLA2 inhibitor bromophenacyl bromide efficiently prevented the cell death in a concentration-dependent manner. Similar results were obtained for the selective cPLA2 inhibitor AACOCF3. However, the Ca 2+ -independent PLA2 inhibitor bromoenol lactone and the secretory PLA2 inhibitor LY311727 were virtually ineffective. These results suggest that cPLA2 plays a causative role in the neuronal death following OGD exposure. Thus, the present study may provide novel therapeutic targets for the development of neuroprotective agents.

1. We investigated the possible involvement of phospholipase A(2) (PLA(2)) and its products in long-term potentiation (LTP) in the CA1 neurotransmission of rat hippocampal slices. 2. Inhibitors of Ca(2+)-independent PLA(2) (iPLA(2)) prevented the induction of LTP without affecting the maintenance phase of LTP whereas Ca(2+)-dependent PLA(2) inhibitors were virtually ineffective, which suggests a pivotal role of iPLA(2) in the initiation of LTP. 3. We then investigated the effect of docosahexaenoic acid (DHA) and arachidonic acid (AA) on BEL (bromoenol lactone, an iPLA(2)-inhibitor) -impaired LTP, and found that either DHA or AA abolished the effect of BEL. However, DHA did not restore BEL-attenuated LTP when applied after the tetanus. DHA per se affected neither the induction nor maintenance of LTP. Linoleic acid had no effects, either. 4. These results suggest that DHA is crucial for the induction of LTP and that endogenously released DHA during tetanus is sufficient to trigger the formation of LTP.

In Suncus murinus the ultrapotent capsaicin analogue resiniferatoxin (RTX) induced an emetic response in the dose range 1–1000 μg kg−1, s.c. The latency was inversely related to dose and ranged from 41.2±4.4 min. (1 μg kg−1, s.c.) to 2.7±0.6 min. (1000 μg kg−1, s.c.). The emetic response to RTX (10 or 100 μg kg−1, s.c.) was blocked or markedly reduced by pre-treatment with RTX (100 μg kg−1, s.c.), 8-OH-DPAT (100 μg kg−1, s.c.), morphine (2 mg kg−1, s.c.), neonatal capsaicin (100 mg kg−1, s.c.) and the NK1 receptor antagonist CP-99,994 (10–20 mg kg−1, s.c.) but not by the 5-HT3 receptor antagonist tropisetron (200 μg kg−1, s.c.). RTX (100 μg kg−1, s.c.) induced c-fos-like immunoreactivity in the area postrema and parts of the nucleus tractus solitarius. This pattern is consistent with the proposal that the emetic effect is mediated via one or both of these structures and an involvement of substance P is discussed. RTX (10 and 100 μg kg−1, s.c.) had broad-spectrum antiemetic effects in Suncus as indicated by its ability to block or markedly reduce the emetic response to motion (1 Hz, 4 cm lateral, 10 min.), cisplatin (20 mg kg−1, i.p.), intragastric copper sulphate (40 mg kg−1, p.o.), nicotine (10 mg kg−1, s.c.) and RTX (100 μg kg−1, s.c.) itself. It is proposed that the site of the anti-emetic effect is in the nucleus tractus solitarius and mechanisms involving the modulation of substance P release are discussed. The general utility of Suncus for investigations of vanilloid receptors is reviewed in the light of the exquisite sensitivity of the emetic reflex in this species to resiniferatoxin. British Journal of Pharmacology (2000) 130, 1247–1254; doi:10.1038/sj.bjp.0703428

Tributyltin (TBT), an assumed endocrine-disrupting chemical, is widely known to show harmful effects in invertebrates including the dioecious snails. As for mammals, there are several reports concerning TBT toxicology, but few indications about general pharmacology of TBT. In the present study, we comprehensively examined the pharmacological effects of TBT both in vivo and in vitro. TBT (0.3 or 1.0 mg/kg) attenuated the small intestinal propulsive activity measured by the charcoal method in vivo, and caused concentration-dependent relaxation of isolated guinea-pig ileum in vitro (1.0x10(-8)-3.0x10(-6) M). TBT induced concentration-dependent relaxation of guinea-pig trachea, which was not inhibited by pre-treatment with a beta-adrenoceptor antagonist. TBT caused a concentration-dependent contraction of rat aortae, and also evoked endothelium-dependent relaxation in the presence of an alpha-adrenoceptor antagonist. The relaxation was inhibited by a muscarinic receptor antagonist. TBT reduced the electrically evoked, sympathetic contractile responses of rat vas deferens, which were slightly prevented by an alpha(2)-adrenoceptor antagonist. These results suggest that TBT possesses diverse pharmacological properties in mammalian organs.

Murine neuroblastoma cell line Neuro-2A cells and rat brain astrocytes showed a dose-dependent increase in intracellular Ca2+ in response to bradykinin, when assessed by a single cell image analyzing system. The Ca2+ increase in Neuro-2A cells by bradykinin was also examined by a suspension fluorescent assay using fura-2 loading. The Ca2+ increase in both cases was suppressed by a bradykinin B2 receptor antagonist, Hoe 140, but not by a B1 receptor antagonist, des-Arg-Hoe 140, suggesting that the effect occurred via specific B2 receptor activation. RT-PCR for bradykinin B2 receptor mRNA showed that both Neuro-2A cells and the astrocytes expressed B2 receptor mRNA. Binding of [3H]bradykinin to Neuro-2A cells was assessed, and a specific binding constant of 0.75 nM was determined. Furthermore, the increase in [Ca2+]i by bradykinin could be caused by a release of Ca2+ from storage sites in the endoplasmic reticulum, since thapsigargin and U-73122 attenuated the effect of bradykinin in Neuro-2A as well as in astrocytes. These results indicate that both astrocytes and neuroblastoma Neuro-2A cells stimulated by bradykinin could express a bradykinin B2 receptor-mediated intracellular Ca2+ increase leading to signal transduction.

A new 5-HT3 receptor ligand, CP2289, was synthesized and pharmacologically tested. Although CP2289 inhibited the Bezold-Jarisch reflex, it contracted the excised ileal muscle of mice, rats and guinea pigs. This response may reflect a partial agonist character of CP2289 in the gut. In vivo antiemetic and gastric emptying tests gave similar results.

Conditioned fear is extinguished if a conditioned animal receives the conditioned stimulus without an unconditioned stimulus. The extinguished fear response can be reinstated after the animal experiences a mild unconditioned stimulus. Although extensive studies on the neuronal circuitry and neurochemical mechanisms leading to fear acquisition and extinction have been carried out, few studies have focused on reinstatement. In this study, we investigated the effects of N-methyl-D-aspartic acid receptor (NMDAR) antagonists, protein synthesis inhibitors, cannabinoid receptor type 1 (CB1R) antagonists, and benzodiazepine on reinstatement of conditioned fear in mice. An intraperitoneal injection of the NMDAR antagonist MK-801 or the protein synthesis inhibitor anisomycin before the reminder shock attenuated fear reinstatement tested the next day. However, anisomycin had no effect on fear reinstatement tested 2 h after the reminder shock. CB1R antagonists, SR141716, and a benzodiazepine, diazepam, had no effect on fear reinstatement. These results suggested that NMDAR and protein synthesis-dependent plasticity contributed toward the reinstatement of conditioned fear and that protein synthesis was involved in consolidation of reinstated fear.

This paper is the first to describe aspects of the mechanics of retching in the insectivore Suncus murinus (house musk shrew) and in an animal of such a small size (approximately 50 g). In anaesthetised animals using the novel stimulus of mechanical stimulation of the upper gastrointestinal tract as the provocative stimulus the frequency of retching was found to be about 4 retches/s, a much higher frequency than in other species (dog, cat, ferret). These studies show that quantification of retching in Suncus cannot be undertaken using direct observation. The temporal pattern of the emetic response was characterised in conscious Suncus using motion (1 Hz, 5 min) and nicotine (20 mg/kg s.c.). The ultrapotent capsaicin analogue resiniferatoxin (100 micrograms/kg s.c.) was discovered to be highly emetic and comparative studies showed that nicotine and resiniferatoxin induced the most intense responses with episodes (retches and a vomit) occurring every 10-15 s. The retching response to mechanical stimulation in the anaesthetised Suncus was not blocked by a 5-HT3 receptor antagonist (granisetron, 1-5 mg/kg s.c.), a tachykinin NK1 receptor antagonist (CP-99,994 20 mg/kg s.c. dihydrochloride salt (9+) -(2S,3S)-3-(2-methoxybenzylamino)-2-phenylpiperidine) or morphine (2 mg/kg s.c.) but was blocked by the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT 100 micrograms/kg s.c.). Suncus appears to be a suitable animal in which to study the pharmacology of the emetic response to mechanical stimulation of the gut. The results are discussed in the light of studies of the pharmacology of emesis in other species.

Neurons can form a synaptic network in culture and show spontaneous oscillation of intracellular Ca2+ concentration ([Ca2+]i). In the present study, spontaneous oscillation of [Ca2+]i was characterized in cultured hippocampal neurons. The oscillation was blocked completely by tetrodotoxin, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and nicardipine, while DL-2-amino-5-phosphonovaleric acid (APV) showed only a partial depression of the increase in [Ca2+]i. These results suggest that the oscillation in [Ca2+]i is mainly mediated by non-N-methyl-D-aspartate (NMDA) type glutamatergic transmission. The oscillation of [Ca2+]i may be a good model for analyzing glutamatergic transmission and synapse formation.

In this review, I have attempted to provide an overview of the pathways by which cytotoxic drugs induce emesis. The mechanisms of serotonin 5-HT3 antagonists and new antiemetics are also discussed. Old data especially from the experiments employing area postrema ablation must be re-evaluated because it is likely that the operation has damaged other important nucleus in the brain stem. Therefore, the concept of the chemoreceptor trigger zone and "vomiting center" proposed by Borison et al. in 1950s is questionable. Nausea and vomiting caused by cytotoxic drugs have been a serious problem in anti-cancer therapy and prompted lots of scientists to find the mechanism and to develop antiemetics. Effectiveness of 5-HT3 antagonists were shown in late 1980s, and now they are clinically available. I have investigated their mechanisms using Suncus murinus and proposed that the pathways by which cisplatin, one of most emetogenic drugs, induces emesis are as follows: 1) cisplatin is converted to an active metabolite(s), 2) the metabolite(s) somehow produces oxygen free radicals in the enterochromaffin cells, 3) the free radicals release serotonin, 4) the released serotonin stimulates 5-HT3 receptors located on the vagus afferents, 5) impulses are transmitted to the brain stem, or emetic pattern generator and initiate emetic reflex. Therefore, scavengers of free radicals and antioxidants can be a new type of antiemetic drug.

Effects of oral administration of cryomilled powder of the soft-shelled turtle (SST powder) on forced-exercise performance and stress-induced reduction in sexual and learning behavior were studied in mice. In the fatigue-preventive experiment, the SSt powder was administered orally to mice which were forced to climb a descending rope for 60 min a day. The treatment significantly improved the daily performance of the mice. In the stress recovery experiment, male mice were hanged from the tail and forced to swim using their fore paws for 30 to 60 min everyday. Stress significantly reduced sexual behavior and memory retrieval (step down test). Daily oral administration of the SST powder significantly attenuated this decline in sexual behavior. However, the effects on memory retrieval were not statistically significant. These results suggest that administration of SST powder attenuates fatigue and accelerates recovery from stress in mice.

1. The effects of (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOl), a 5-hydroxytryptamine 5-HT2A/5-HT2C receptor agonist, on motion- and cisplatin-induced emesis were studied in Suncus murinus. Subcutaneous injection of DOl, 30 min prior to the emetic stimuli, dose-dependently blocked the emesis induced by motion sickness and cisplatin (20 mg kg-1, i.p.) with estimated ID50 values of 640 and 780 micrograms kg-1, respectively. 2. alpha-Methyl-5-hydroxytryptamine (alpha-Me-5-HT), a peripheral 5-HT2A/5-HT2C receptor agonist, had no effect on motion- and cisplatin-induced emesis. 3. The antiemetic effects of DOl on motion- and cisplatin-induced emesis were attenuated by preadministration of ketanserin, a selective 5-HT2A receptor antagonist. 4. The present results suggest an inhibitory role for central 5-HT2 receptors in the emetic reflex mechanism and that a 5-HT2 receptor agonist may be a useful tool to investigate the involvement of 5-HT receptors in the emetic reflex.

Cannabinoid agonists impair hippocampus-dependent learning and memory. Using mouse hippocampal slice preparations, we examined the effect of anandamide, an endogenous cannabinoid, on sharp wave–ripple (SW-R) complexes, which are believed to mediate memory consolidation during slow-wave sleep or behavioral immobility. Anandamide reduced the frequency of SW-Rs recorded from the CA3 region, and this effect was abolished by AM251, a cannabinoid CB1–receptor antagonist. We further addressed the action of anandamide using a functional multineuron calcium imaging technique. Anandamide reduced the firing rate of hippocampal neurons as well as disrupted the temporal coordination of their firings during SW-R. Keywords:: hippocampus, local field potential, memory consolidation, endocannabinoid, type 1 cannabinoid (CB1) receptor

To clarify the role of cholinergic neurons in the amygdala on learning and memory, scopolamine was injected topically into the bilateral amygdala of mice, and the ability to perform two types of passive avoidance tasks (step-through and step-down) was investigated. On the first day mice performed the learning trial and on the second day their retention was tested. Scopolamine (0.5 or 1 microgram/site) was injected bilaterally into the amygdala 30 min before or immediately after the learning trial or 30 min before the testing trial. Scopolamine impaired the performance ability of the mice dose-dependently only when it was injected 30 min before the learning trial. The results coincided well with the effect of scopolamine injected intraperitoneally. Taken together, these results suggest that the site of action for scopolamine to cause anterograde amnesia is the amygdala, and that cholinergic neurons projecting to the amygdala play an important role in memory acquisition in the two passive avoidance tasks.

This paper reports that vasopressin is emetogenic in the house musk shrew Suncus murinus. Either intravenous or intracerebroventricular administration of vasopressin caused vomiting within a few minutes. The ED50 of intravenous vasopressin was as high as 4.67 microg/kg, whereas intracerebroventricularly injected vasopressin was effective at a low dose of 20 ng/brain. The emetogenic target of vasopressin may therefore be present in the central nervous system. We propose the Suncus as a useful animal for investigation of vasopressin-mediated emesis, including motion sickness.

Cerebral ischemia causes the depletion of oxygen and nutrition from brain tissues, and when persistent, results in irreversible damage to the cell function and survival. The cellular response to ischemic conditions and its mechanisms have been investigated widely in in vivo and in vitro experimental models, yet no study has addressed the response of a whole neuronal network to energy deprivation with the single-cell resolution. Observations at the level of network are necessary, because the activity of individual neurons is nonlinearly integrated through a network and thereby gives rise to unexpectedly complex dynamics. Here we used functional multineuron calcium imaging (fMCI), an optical recording technique with high temporal and spatial resolution, to visualize the activity of neuron populations in hippocampus CA1 region under ischemia-like conditions ex vivo. We found that, although neurons responded to oxygen and glucose deprivation with an increase in the event frequency, they maintained an asynchronous network state. This is in contrast with other well known pathological states, in which the network hyperexcitability is usually accompanied by an increase in synchrony. We suggest that under ischemic conditions, at least to some time point, the neuronal network maintains the excitatory and inhibitory balance as a whole, whether actively or as a consequence of the cellular response to energy deprivation.

The participation of free radicals in cisplatin-induced emesis was investigated in the house musk shrew, Suncus murinus. Thiobarbituric acid (TBA) values, which indicate the degree of lipid peroxidation, in brain, liver and small intestine were increased significantly 60 min after the treatment with cisplatin (20 mg/kg, i.p.). Moreover, cisplatin (20 mg/kg, i.p.)-induced emesis was prevented by intraperitoneal injection of N-(2-mercaptopropionyl)glycine (MPG), a radical scavenging agent, with ID50 value of 130 mg/kg. However, MPG did not block the emesis induced by copper sulfate (40 mg/kg, p.o.), veratrine (0.5 mg/kg, s.c.) or serotonin (10 mg/kg, i.p.). We also investigated the effects of superoxide dismutase conjugated to polyethylene glycol and catalase, but the number of vomiting episodes and latency did not change significantly when these agents were intraperitoneally injected 30 min prior to or 20 min after the administration of cisplatin. MPG did not affect the antitumor effect of cisplatin tested in vitro. These results suggest that free radicals mediate emesis caused by cisplatin and that radical scavengers may become a new class of prophylactic drug against cancer-chemotherapeutic drug-induced emesis.

Effects of aniracetam on whole-cell calcium currents were studied in acutely isolated neuronal cells from postnatal rat ventromedial hypothalamus. There were three types of inward calcium currents, one low-threshold transient current and two high-threshold sustained currents. The nicardipine sensitive L-type current was activated at -20 mV or more depolarized potentials, and the omega-conotoxin sensitive N-type current was recorded at more positive potentials than the L-type. Aniracetam inhibited the N-type current in a dose-dependent manner without affecting the other two types of calcium currents. The effect appeared soon after the addition and lasted for several minutes during washing. Since the N-type current is thought to regulate the release of transmitters, the inhibitory effect may contribute to the nootropic property of aniracetam by modifying the neurotransmission.

In this review, I have attempted to provide an overview of the pathways by which cytotoxic drugs induce emesis. The mechanisms of serotonin 5-HT3 antagonists and new antiemetics are also discussed. Old data especially from the experiments employing area postrema ablation must be re-evaluated because it is likely that the operation has damaged other important nucleus in the brain stem. Therefore, the concept of the chemoreceptor trigger zone and "vomiting center" proposed by Borison et al. in 1950s is questionable. Nausea and vomiting caused by cytotoxic drugs have been a serious problem in anti-cancer therapy and prompted lots of scientists to find the mechanism and to develop antiemetics. Effectiveness of 5-HT3 antagonists were shown in late 1980s, and now they are clinically available. I have investigated their mechanisms using Suncus murinus and proposed that the pathways by which cisplatin, one of most emetogenic drugs, induces emesis are as follows: 1) cisplatin is converted to an active metabolite(s), 2) the metabolite(s) somehow produces oxygen free radicals in the enterochromaffin cells, 3) the free radicals release serotonin, 4) the released serotonin stimulates 5-HT3 receptors located on the vagus afferents, 5) impulses are transmitted to the brain stem, or emetic pattern generator and initiate emetic reflex. Therefore, scavengers of free radicals and antioxidants can be a new type of antiemetic drug.

The effects of orally administered soft-shelled turtle powder on experimental liver injury induced by carbon tetrachloride were studied. Activities of transaminases and lactate dehydrogenase, content of albumin in serum, total protein, triglyceride and hydroxyproline in liver and histopathological assessment were used as indices of liver injury. Daily oral administration of 100 and 500 mg/kg soft-shelled turtle powder for 6 weeks did not prevent the acute injury induced by a subsequent single injection of carbon tetrachloride. However, administration of soft-shelled turtle powder for 6 weeks during chronic treatment with carbon tetrachloride significantly and dose-dependently ameliorated the liver injury. The increases in glutamic-pyruvic transaminase, glutamic-oxaloacetic transaminase and lactate dehydrogenase in serum were attenuated, and there was a noticeable improvement of serum album and liver total protein content in rats treated with the powder. In addition, histopathological observation demonstrated a decrease in fatty degeneration in the liver of rats treated with the powder. None of the indices of liver injury was significantly affected by 6-week oral administration of soft-shelled turtle powder in non-carbon tetrachloride-treated rats. The results suggest that repeated administrations of soft-shelled turtle powder have a therapeutic effect on chronic liver injury, by improving liver function and protein synthesis.

The effects of chronic oral administration of cryomilled powder of the soft-shelled turtle (SST powder) on blood pressure, serum catecholamine contents and vascular reactivity of spontaneously hypertensive rats (SHR) and normotensive Wistar Kyoto rats (WKY) were studied. The SST powder was administered orally 20–500 mg/kg/day for 9 weeks. The treatment slowly but significantly reduced the blood pressure increase in SHR. However, the SST powder did not affect the blood pressure in WKY. Heart rate and wet heart weight were not affected by the SST powder treatment. Serum content of catecholamine and aldosterone and the plasma renin activity were also unaffected by the chronic administration of the SST powder. The increase in vascular resistance of isolated mesenteric vascular beds from SHR was significantly reduced by the chronic treatment of SST powder. Vascular reactivity to norepinephrine was also smaller in the SST powder-treated group. The treatment did not affect vascular resistance or reactivity in WKY. These results suggest that chronic oral administration of the SST powder reduces the development of hypertension by decreasing the peripheral vascular resistance and reactivity. The SST powder may be useful for the treatment of hypertension.

Although primary cultures of neurons are essential methods for cell biological and pharmacological researches, many animals must be sacrificed for each experiment. Here we introduce a novel system to cryopreserve hippocampal granule cells (GCs) prepared from postnatal rats. Being thawed after as long as 60 days of cryopreservation, GCs expressed the mature neuronal marker MAP-2 and elongated single tau-1-positive axons and multiple tau-1-negative dendrites. These properties closely resembled intact GCs in primary cultures, providing the advantage of being able to repeatedly prepare stable cultures with a single sacrifice of animals. Keywords:: dentate gyrus, primary culture, Prox1

Controlling axon and dendrite elongation is critical in developing precise neural circuits. Using isolated cultures of dentate granule neurons, we succeeded in simultaneously monitoring the behaviors of axonal and dendritic outgrowth. Our previous study shows that cAMP contributes differentially to Sema3F-induced responses of axons and dendrites, but we report here that the cGMP modulation does not have such a striking axo-dendritic difference. Treatment with Sema3F induced collapse of about 90% growth cones, and pretreatment with 1 muM LY83583, an inhibitor of soluble guanylyl cyclase, partially alleviated the collapse of both axons and dendrites. Thus, unlike cAMP, cGMP modulates axonal and dendritic extension in a similar manner.

It is difficult to comprehend the entorhino-hippocampal information processing using acute transverse hippocampal slice, because the dorsally inclined connections of the entorhino-hippocampal projections can be damaged easily. Therefore, we investigated the spatial-temporal propagation in organotypic cultures of the hippocampus attaching to the entorhinal cortex using a real-time optical recording system with a voltage-sensitive dye and suitability as an in vitro model. Real-time imaging demonstrated that the stimulation of the perforant pathway induced excitatory propagation in trisynaptic pathway of the hippocampus and sequentially in the layer V from the medial to the lateral entorhinal cortex. The horizontal propagation from the lateral to the medial site was also seen after the stimulation of the lateral entorhinal cortex. The analysis of the entorhino-hippocampal organotypic culture would contribute to understanding of the mechanism of learning and memory.

The prophylactic effect of serotonin uptake inhibitors, imipramine and fluoxetine, against motion sickness was investigated in Suncus murinus. Imipramine (s.c.) and fluoxetine (i.p.) inhibited motion-induced emesis dose dependently with ID50 values of 1.7 and 26 mg/kg, respectively. The results suggest that increasing the concentration of serotonin in the synaptic cleft can prevent motion-induced emesis and that serotonin uptake inhibitors are effective as anti-motion sickness drugs.

Amyloid beta-protein (Abeta), a putative pathogenic endotoxin involved in Alzheimer's disease, induces redistribution of glutamate transporters in astrocytes and promotes their pump activity. Because the transporters are assumed to protect neurons against excitotoxicity by removing extracellular glutamate, we hypothesized that Abeta alters the vulnerability of neurons to glutamate. Cerebrocortical neuron-astroglial co-cultures were exposed to glutamate, the concentration of which was selected so that only 20% of the neurons exhibited degeneration. When cultures were pre-treated with Abeta, exposure to the same "mild" glutamate concentration failed to damage neurons. The Abeta-induced protection was abolished by a glial glutamate transporter inhibitor. Thus, Abeta can alleviate excitotoxicity through glutamate transporter activity. The present results may challenge prevailing concepts that Abeta-induced neuron loss causes Alzheimer's dementia and also provide practical insights into neuro-glial interactions in glutamate toxicity.

The nucleus of solitary tract (NTS) contains diverse neural circuits responsible for basic vital functions. We examined the effect of serotonin (5-HT) on hyperpolarization-activated current (I(h)) in neurons acutely isolated from caudal, medial and rostral parts of the NTS. Caudal and medial NTS neurons showed a large amplitude of I(h) compared with rostral neurons. In these neurons, perfusion with 5-HT potentiated Ih amplitude in a concentration-dependent manner. The effect of 5-HT was blocked by NAN-190, a 5-HT1A receptor antagonist. Thus, 5-HT1A receptors may regulate I(h) channel activity in caudal and medial NTS neurons.

It is well demonstrated that acute or chronic stress leads to reduction of learning ability. Lipopolysaccharide (LPS), a component of the outer membrane of gram-negative bacteria, induces profound physiological and behavioral changes, including fever, decrease in food motivation, and decrease in social behavior. These changes might be interpreted as an acute stress reaction to the LPS. In the present study, therefore, we investigated the effects of LPS (400-800 microg/kg, i.p.) on spatial learning performances using C57BL/6J male mice. In the Morris water-maze task, spatial learning performances were examined in six trials of training for two consecutive days. LPS-treated mice took a longer time to reach the hidden platform than control mice (F(1,60)=4.80801, P0.05 at 600 microg/kg). In addition, injection of LPS decreased the percent of correct choices in the Y-maze test (P0.05 at 800 microg/kg). LPS, however, did not alter the body weight, grip tone, motor activity or swimming speed. Taken together, these results indicate that LPS treatment specifically impaired spatial learning performances.

The effects of ferric chloride and deferoxamine, an iron chelator, on cisplatin-induced emesis were studied in Suncus murinus . Pretreatment with ferric chloride (FeCl 3 , 16–64 mg/kg) increased the number of 20 mg/kg cisplatin-induced vomiting episodes. Deferoxamine (256 mg/kg) significantly reduced the number of vomiting episodes and prolonged the latency. These results further support the involvement of free radicals in the generation of cisplatin-induced emesis.

Rationale: Opiate-dependent subjects experi- ence severe depression as one of the subjective symp- toms during withdrawal. No experimental work, how- ever, has focused on the ability of opiate-withdrawal to produce depression-like behavior in dependent animal. Objectives. We therefore investigated whether withdraw- al from chronic morphine treatment affects immobility in forced swimming test in rats. Methods: Morphine was administered in a dose escalation fashion using doses ranging from 20 to 140 mg/kg twice daily for 14 days, followed by 1-6 days of withdrawal, and their duration of immobility was assessed. Results: After the last mor- phine treatment, an increase in immobility occurred late on day 3 and persisted to at least day 6 of withdrawal without any change in ambulatory activity. Conclusions: The results suggest that the morphine withdrawal re- sulted in prolonged enhancement of depression-like be- havior in drug-dependent laboratory animals.

We investigated the effect of tributyltin (TBT), an endocrine-disrupting chemical, on the morphology and viability of cultured rat cortical astrocytes. Cultured astrocytes exhibited smooth and planiform morphology under normal conditions. Following exposure to TBT, however, they showed rapid morphological changes that are characterized by asteriated cell bodies and process formation in a time- and concentration-dependent manner. Higher concentrations of TBT produced progressive cell death of the astrocytes. In serum-free medium, TBT at a concentration as low as 200 nM induced the stellation. Pharmacological studies revealed that the morphological changes were alleviated by application of diverse free radical scavengers or antioxidants such as catalase, superoxide dismutase, Trolox, ascorbic acid and N-acetyl-L-cysteine, suggesting that TBT-induced stellation is caused by oxidative stress involving free radicals, particularly reactive oxygen species. Furthermore, we found that the astrocyte stellation was abolished by treatment with inhibitors of phospholipase C, mitogen-activated protein kinase kinase or tyrosine phosphatase. The data suggest that TBT causes the stellation through intracellular signaling cascades rather than its non-specific toxicity. These findings provide an important insight for reconciling the problems in assumed aversive actions of this environmental pollutant for mammals.

Electrical signals in rat hippocampal slices were optically monitored using a voltage-sensitive dye to determine whether extracellular ATP exhibits direct effects through its own receptors or indirect effects after its hydrolysis to adenosine. The dentate gyrus was stimulated and electrical signals in the CA1 and the CA3 region were analyzed. The signals were divided into two components: a transient component peaking within 10 ms (fast component) and a subsequent sustained component (slow component). ATP (10 to 100 μM) inhibited both the fast and the slow components in the CA1 region by about 30% and 70%, respectively. ADP, AMP and adenosine also inhibited the fast and the slow components. The inhibition by ATP was antagonized by aminophylline and other adenosine receptor antagonists, and by α,β-methylene ADP, an inhibitor of 5′-nucleotidases. These results suggest that extracellular ATP inhibits neuronal electrical signals in hippocampal slices after its metabolism to adenosine.

The emetic (retching and vomiting) reflex is an important component of the body's defence system against accidentally ingested toxins and emesis is also a common symptom of disease and a side-effect of a number of pharmacological therapies. The development of the reflex has been the subject of few systematic studies. The aim of this study was to characterise the development of the emetic reflex in Suncus murinus (the house musk shrew) using emetic stimuli acting via three different afferent pathways: motion via the vestibular system, pyrogallol via abdominal vagal afferents and resiniferatoxin (a capsaicin analog) via the brainstem. The emetic reflex was not present to any stimulus prior to postnatal day 10 but the onset of the response to motion lagged behind that to the other stimuli in not being present until postnatal day 15. Body weight was not a determinant of the presence of the reflex. It is proposed that the delayed presence of the emetic reflex in Suncus makes it an ideal species in which to investigate factors regulating its development.

Neurotrophic effects in vitro have been generally related to promotion of differentiation, maturation and survival, but little is known about the effect on neuronal circuit formation. The organotypic culture system would be an available technique to investigate neuronal circuit formation and neuronal cell-cell interactions. As we reported previously, an optical recording system is a useful technique to comprehend neuronal activities and circuit from multi-points simultaneously. In this study, we investigated whether continuous application of basic fibroblast growth factor (bFGF or FGF-2) and brain-derived neurotrophic factor (BDNF) inhibited neuronal cell death induced by serum-deprivation in organotypic culture using propidium iodide staining, and we analyzed effects of bFGF and BDNF on the formation of neuronal circuits using the optical recording system. Continuous application of bFGF or BDNF significantly protected the slices from neuronal death. Optical recording also demonstrated that addition of 10 ng/ml bFGF or 50 ng/ml BDNF enhanced optical signals in all hippocampal areas significantly. These data strongly suggest that bFGF and BDNF promote the formation of neuronal circuits as well as survival and that optical recording of organotypic hippocampal slices would be a useful technique that enables us to analyze neuronal circuit formation easily.

In the CA1 region of rat hippocampal slices, H 2 O 2 (0.294–2.94 mM) caused initial augmentation, and subsequent long-lasting depression, of population spikes and excitatory postsynaptic potentials. The effect of H 2 O 2 may not be mediated by its degradation product, hydroxyl radicals, because an iron chelator deferoxamine did not block the effect. A catalase inhibitor 3-amino-1,2,4-triazole only modestly attenuated the initial augmentation, suggesting that the effect of H 2 O 2 is not attributable to catalase-dependent O 2 generation, either. An N -methyl- d -aspartate receptor antagonist dl -2-amino-5-phosphonovaleric acid had no influence on the effect of H 2 O 2 , whereas a γ -aminobutyric acid type A receptor channel blocker picrotoxin attenuated long-lasting depression, indicating that γ -aminobutyric acid-mediated inhibition is altered during the depression phase. The initial augmentation but not subsequent depression was attenuated by a phospholipase A 2 /C inhibitor 4-bromophenacyl bromide, suggesting the involvement of lipid signaling molecule(s) in the enhancement of excitatory synaptic transmission. These results suggest that H 2 O 2 regulates hippocampal synaptic transmission via multiple mechanisms.

In order to elucidate possible male/female differences in emesis, the effects of various emetogenic drugs (cisplatin, copper sulfate, veratrine, nicotine, serotonin) and motion stimulus were compared between male and female Suncus murinus. Cisplatin (IP), nicotine (SC), veratrine (SC) and copper sulfate (PO) induced dose-dependent emesis in either sex, and there was no apparent difference in estimated ED50 values. However, male animals tended to be more susceptible to serotonin-induced emesis. The ID50 values for tropisetron, a 5-HT3 receptor antagonist, to block serotonin-induced emesis were also similar between male and female animals. However, tropisetron was less effective against cisplatin-induced emesis in females. Therefore, cisplatin may release more serotonin to induce emesis in females. Reciprocal shaking (horizontal oscillation 40 mm, frequency 0.5 to 2.0 Hz, duration 5 min) induced more frequent emesis in male animals, and the latency to the first vomit was shorter in males than in females. These results suggest that there is substantial sex-dependent difference in the emetic responses and male animals are in general more susceptible. These results are discussed in the light of similar studies in man.