Calcium entry into nerve termini via voltage gated calcium channels is an essential step in neurotransmission. Consequently, second messenger regulation of calcium channel activity modulates synaptic activity. It has been suggested that calcium channels must physically couple to the release machinery, and a physical interaction between a synaptic protein interaction (synprint) site contained within mammalian presynaptic calcium channels and synaptic proteins such as syntaxin 1, SNAP-25, and synaptotagmin has been demonstrated. Interestingly, synaptic calcium channels in invertebrates lack this region. In invertebrates, synaptic transmission is instead dependent on a presynaptic calcium channel splice variant that can physically associate with the adaptor proteins Mint-1 and CASK. We suggest that in the absence of a synprint region, these proteins may localize calcium channels to the synaptic release machinery. The interactions between synaptic proteins and mammalian N-type calcium channels serves to regulate calcium channel activity directly, as well as indirectly by altering second messenger regulation of the channels. This provides for a feedback mechanism that allows the fine-tuning of calcium channel activity during various steps in neurotransmitter release. This does not occur with invertebrate synaptic calcium channel homologs, suggesting that the regulation of calcium channel activity by synaptic proteins is a mechanism unique to vertebrates.

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It has been recognized that sympathetic neurons release several transmitters but mainly adenosine 5'-triphosphate (ATP), noradrenaline, and neuropeptide Y (NPY). Recently, we reported that periarterial nerve electrical stimulation (PNS) produced biphasic vasoconstrictions consisting of an initial transient, predominantly P2X-purinoceptor-mediated constriction followed by a prolonged, α₁-adrenoceptor-mediated one in canine isolated splenic arteries. In this article, we tried to analyze the effects of several selective key drugs that influence the PNS-induced responses, and we functionally showed sympathetic transmitter releasing mechanisms by pharmacological analysis using purinergic, adrenergic, and NPYergic agonists and antagonists.

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Porcine cerebral arterial strips denuded of the endothelium responded to transmural electrical stimulation (5 Hz for 40 s) with a relaxation, which was abolished by tetrodotoxin and N ^G-nitro-L-arginine, a NO synthase inhibitor. Lowering the temperature of the bathing media from 37°C to 33°C or 25°C potentiated the response to nerve stimulation, but did not affect relaxations induced by NO applied exogenously. Hypoxia suppressed the stimulation-induced relaxation at 37°C, but hypothermia blunted the inhibitory effect of hypoxia in a temperature-dependent manner. It is concluded that hypothermia augments vasodilatation associated with nitroxidergic (nitrergic) nerve activation possibly by increasing the production of NO from L-arginine and, in addition, prevents impairment of NO production by hypoxia. These mechanisms likely explain how hypothermia protects nerve cells against hypoxia. Inhibitions of cyclic GMP phosphodiesterase and of superoxide production by hypoxia do not seem to participate in the action of hypothermia. Mechanisms underlying its protective action remain to be ascertained.

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There is evidence indicating that dysregulation of coordinated interactions of the cortical-limbic circuitry is associated with anxiety and mood disorders. Our previous study has reported that an enhancement of long-term plasticity in the “limbic-cortical” pathway produced by repeated treatments with fluvoxamine may be involved in the clinical effects of a selective serotonin (5-HT) reuptake inhibitor (SSRI). Here we assessed the effects of single and repeated treatments with fluvoxamine on the synaptic transmission and plasticity in the “cortical-limbic” pathway in vivo. The evoked potentials in the basolateral amygdaloid complex (BLA) by stimulation of the medial prefrontal cortex (mPFC) in halothane-anesthetized rats were recorded. Single administration of fluvoxamine (10 and 30 mg/kg, i.p.) enhanced the efficacy of synaptic transmission at the mPFC-BLA synapses dose-dependently. The enhanced synaptic efficacy induced by 30 mg/kg fluvoxamine was suppressed after long-term administration of fluvoxamine (30 mg/kg per day × 21 days, orally). Repeated treatments with fluvoxamine affected short-term, but not long-term, synaptic plasticity in the mPFC-BLA pathway. These findings indicate that the 5-HTergic system contributes to modulation of synaptic changes in this pathway. Our results also suggest that different changes in synaptic properties in cortical-limbic communications induced by repeated treatments with fluvoxamine may be associated with therapeutic effects of SSRI.

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It is well established that spasm of an arterial and venous graft conduit may occur during harvesting or after coronary artery bypass grafting (CABG). The antivasoconstrictor effect of levcromakalim and rilmakalim, K⁺ channel openers (KCOs), was studied in isolated human internal mammary artery (HIMA) and human saphenous vein (HSV) prepared for CABG. HIMA and HSV rings were contracted by electrical field stimulation (EFS, 20 Hz ) or with exogenous noradrenaline (NA). Levcromakalim induced a concentration-dependent and equipotent inhibition of contraction of HIMA and HSV preconstricted by EFS and exogenoulsy applied NA, while rilmakalim produced a stronger inhibition of EFS- than NA-evoked contractions. Glibenclamide, a selective ATP-sensitive K⁺ channel (K_ATP channel) blocker, significantly antagonized levcromakalim-induced inhibition of EFS- and NA-evoked contractions, as well as rilmakalim-induced inhibiton of EFS-evoked contractions on HIMA and HSV. However, glibenclamide failed to antagonize rilmakalim-induced inhibition of NA-evoked contractions. The results suggest that the antivasoconstrictor effect of levcromakalim occurs postsynapticaly by the opening K_ATP channels in the vascular smooth muscle cells. They also suggest that the effect of rilmakalim on EFS-evoked contractions involves K_ATP channels located pre-synaptically. However, the mechanism by which rilmakalim inhibits NA-evoked contraction seems to be K_ATP channel independent and warrants further elucidation.

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Alzheimer’s disease (AD) is characterized by the accumulation of extracellular amyloid-β (Aβ) fibrils with microglia. Recently, there has been great interest in the microglial phagocytosis of Aβ, because the microglial pathway is considered to be one of the Aβ clearance pathways in the brain parenchyma. However, the mechanism of microglial phagocytosis of Aβ is not fully understood and, thus, was investigated in this study. At one minute after exposure to Aβ(1 – 42) (Aβ42), Aβ immunoreactivity was detected at the cell surface of microglia. After 1 h, marked immunoreactivity was observed in the cytosolic vesicles. At 12 h, delayed phagocytosis of fibrillar Aβ42 was also observed with the formation of a large phagocytic cup. The microglial cell shape rapidly changed to an ameboid form during the process of phagocytosis. Although neither neural Wiskott-Aldrich syndrome protein (N-WASP) nor WASP interacting SH3 protein (WISH) immunoreactivity was co-localized with filamentous actin (F-actin) distribution, both WASP family verprolin-homologous protein (WAVE) and Rac1 immunoreactivity was co-localized with F-actin in the lamellipodia of phogocytic microglia. These results suggest that WAVE and Rac1 participate in the phagocytosis of Aβ42 by microglia.

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Although it is well known that histamine induces gastric mucosal lesions in laboratory animals, the fundamental mechanisms remain unclear. In order to further analyze the vascular mechanisms underlying histamine-induced lesions, a new model was developed in the glandular stomach via administration of histamine (40 mg/kg, s.c.) twice to rats with partial gastric vascular occlusion (ligated left gastric artery and vein) also subjected to pylorus ligation. Both antagonists of histamine H₂-receptors (roxatidine and famotidine) and H₁-receptors (epinastine and tripelennamine) significantly inhibited lesion formation at doses that did not inhibit acid secretion. Combined treatment of tripelennamine and famotidine synergistically inhibited lesion formation. Nitro L-arginine methyl ester inhibited lesion development; inhibition was reversed by concomitantly administered L-arginine. Indomethacin, diclofenac, and SC-560 (a selective COX-1 inhibitor), but not rofecoxib (a selective COX-2 inhibitor), significantly inhibited lesion formation. In addition, sodium bicarbonate, pirenzepine, S-0509 (a gastrin/CCK₂ inhibitor), omeprazole, sucralfate, and a prostaglandin analog also significantly inhibited lesion formation. It was concluded that the mechanism by which histamine induces gastric lesions in rats with partial gastric vascular occlusion appears to involve extensive vasodilation resulting from histamine acting on microvasculature histamine H₁- and H₂-receptors, generation of endogenous nitric oxide and prostaglandins, with the presence of gastric acid.

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It has been known that rotenone and 1-methyl-4-phenylpyridinium ion (MPP⁺, a metabolite of MPTP), which inhibit mitochondrial complex I, are useful tools for parkinsonian models in vertebrates such as primates and rodents. Planarian, an invertebrate flatworm, has a high potential for regeneration, and dopamine plays a key role in its behavior. In the present study, we examined a cloned planarian, the GI strain from Dugesia japonica. Planarians that were treated with rotenone or MPTP underwent autolysis and individual death in a concentration- and time-dependent manner. In addition, these effects induced by rotenone or MPTP were inhibited by several antiparkinsonian drugs and caspase inhibitors. These results suggest that the degeneration of planarian dopaminergic system induced by rotenone or MPTP may be mediated through caspase-like activation.

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Involvement of the dopamine D₁ and D₂ receptors in the ventral tegmental area (VTA) in the micturition reflex was investigated using female Sprague Dawley rats under urethane anesthesia. Cystometrograms during continuous infusion of warmed saline into the bladder were recorded. When intervals of bladder contraction became constant, the excitatory amino acid DL-homocysteic acid (DLH) was microinjected into the VTA and changes in the cystometric parameters were observed. The selective D₁ antagonist SCH23390 (0.3 mg/kg) or D₂ antagonist eticlopride (0.1 mg/kg) was subcutaneously injected (s.c.) 15 min prior to the DLH treatment. A low dose of DLH (3 μg) facilitated the micturition reflex, whereas a high dose of DLH (30 μg) inhibited the micturition reflex. However, a middle dose of DLH (10 μg) did not show any effect. The facilitated micturition reflex induced by a low dose of DLH was inhibited by the selective dopamine D₂ antagonist eticlopride, but unaltered by SCH23390, a selective dopamine D₁ antagonist. In contrast, the inhibitory effect induced by a high dose of DLH on the micturition reflex was suppressed by SCH23390 but not eticlopride. These results suggested that the facilitated micturition reflex might be mediated via the dopamine D₂ receptors, while the inhibitory effects on the micturition reflex was mediated via the dopamine D₁ receptors.

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We investigated the acute effects of thiopental anesthesia (4 mg/100 g, i.v.) on plasma glucose, insulin, triacylglycerol, and prolactin levels in rats treated with bromocriptine (BR) (0.4 mg/100 g body wt, i.p., for two weeks). Thiopental anesthesia induced a rapid increase in plasma insulin that was more pronounced in the animals treated with BR (116%, P<0.05). Thiopental anesthesia also produced a 55% decreased in plasma prolactin levels (P<0.01) in control fed rats, and a 22% reduction in plasma triacylglycerol (P<0.05) in both controls and BR-treated rats. We conclude that BR may constitute and additional sympatholytic factor in animals submitted to thiopental anesthesia.

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The present study examines the effect of the cognition enhancer (1R)-1-benzo[b]thiophen-5-yl-2-[2-(diethylamino)ethoxy]ethan-1-ol hydrochloride (T-588) on neuronal injury induced by serum deprivation or amyloid-β protein (Aβ). T-588 protected partially against neuronal injury induced by serum deprivation or Aβ in cultured cortical neurons. T-588 did not affect the phosphorylation of extracellular signal-regulated kinase (ERK) in cortical neurons and SH-SY5Y cells. These results suggest that T-588 has a protective effect in neuronal injury models and the effect is not mediated by an ERK signal pathway.

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The effects of different concentrations of N^G-nitro-L-arginine methyl ester (L-NAME) (0.3, 1, 3, and 10 mM), a non-selective inhibitor of NOS, on the effect of aminophylline on the isometric contraction of the isolated rat hemidiaphragm were investigated. The muscle contractions were induced by direct subtetanic electrical stimulation. Aminophylline (0.36 – 3.60 mM) produced a typical concentration-dependent increase in both parameters of the isometric contraction: tension developed (Td) and the maximum rate of rise of tension (dT/dt max). The second series of additions of aminophylline produced a more pronounced effect. L-NAME (0.3, 1, 3, and 10 mM, 30 min of incubation without stimulation) itself did not change Td and dT/dt max. However, L-NAME (1, 3, and 10 mM) produced a statistically significant potentiation of the effect of aminophylline on Td and dT/dt max.

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Manganese(III)tetrakis(1-methyl-4-pyridyl)porphyrin (MnTMPyP), which has been known as a cell permeable superoxide dismutase mimetic, induced concentration-dependent contraction in rat carotid artery acting directly on smooth muscle. The contractile action was more prominent in the preparation from stroke-prone spontaneously hypertensive rats (SHRSP) compared with that from Wistar Kyoto rats (WKY). It was abolished by the removal of extracellular Ca²⁺ or the application of verapamil. These results suggest that the MnTMPyP-induced contraction is brought about by Ca²⁺ influx through voltage-dependent Ca²⁺ channels (VDCC) and that the difference in VDCC is the cause of the difference in MnTMPyP action between preparations from WKY and SHRSP.

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Flavonoids are widely recognized as naturally occurring antioxidants. Naringin (NG) is one of the flavonoid components in citrus fruits such as grapefruit. Hydrogen peroxide (H₂O₂) causes cytotoxicity through oxidative stress and apoptosis. In this paper, we examined the effects of NG on H₂O₂-induced cytotoxicity and apoptosis in mouse leukemia P388 cells. Cytotoxicity was determined by mitochondrial activity (MTT assay). Apoptosis and DNA damage were analyzed by measuring chromatin condensation and Comet assay (alkaline single cell gel electrophoresis), respectively. H₂O₂-induced cytotoxicity was significantly attenuated by NG or the reduced form of glutathione (GSH), a typical intracellular antioxidant. NG suppressed chromatin condensation and DNA damage induced by H₂O₂. These results indicate that NG from natural products is a useful drug having antioxidant and anti-apoptopic properties.

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