Nitric oxide (NO) release by glutamate and NMDA in the dorsal horn of the spinal cord: an in vivo electrochemical approach in the rat

Glutamate acts as a neurotransmitter of primary afferent messages in the spinal cord. Through glutamatergic mechanisms nitric oxide (NO) is also a potential intermediary in the transmission of sensory messages, particularly nociceptive, at the spinal level. The aim of the present study was, by using electrochemical monitoring of NO, to determine if the activation of glutamatergic transmission, particularly through NMDA receptors, could increase NO production within the dorsal horn of the lumbar spinal cord in the rat. 30 micrometers diameter treated carbon fiber electrodes coated with nickel-porphyrine and Nafion(R), and associated with differential normal pulse voltammetry, have been used in vivo to monitor NO within the dorsal horn of the lumbar spinal cord of decerebrated-spinalized rats. A NO-dependent peak of oxidation current (650 mV vs. Ag-AgCl), remaining stable for up to 3 h (+/-5%) could be detected under basal conditions, which indicates that significant amounts of NO are produced continuously. The non-competitive N-methyl-d-aspartate (NMDA) channel blockers, Ketamine (100 mg kg-1 i.p.) and MK-801 (10 mg kg-1 i.p.), decreased the voltammograms to 70+/-5% and 69+/-2% of controls at 120 min, respectively. Glutamate (10 mM), when directly superfused upon the spinal cord (20 min at 50 microliters min-1) induced a rapid and significant increase of the 650 mV peak, with a maximum at around 90 min (148+/-6% of control) followed by a slow decay (138+/-4% of control at 150 min). This increase could be totally reversed or blocked by i.p. injection of 100 mg kg-1 of Ketamine. NMDA (30 mg kg-1 i.p.) induced a long-lasting increase in the peaks (149+/-11% at 90 min and 162+/-20% at 120 min), which was also fully reversed by Ketamine or MK-801. These results provide in vivo direct evidence of a glutamate- and/or NMDA-induced release of NO at the spinal level, and is discussed in relation to the glutamatergic transmission of primary afferent messages.     

Voltammetric studies of Cu-adriblastina complex and its effect on ssDNA-adriblastina interaction at<Emphasis Type="Italic">in situ</Emphasis> mercury film electrode

Adriblastina, a cancerostatic anthracycline antibiotic, causes considerable oxidative damage to DNA molecules. The interaction of this compound with DNA was investigated using Osteryoung square wave stripping voltammetry (OSWSV) and cyclic voltammetry (CV) at an in situ mercury film electrode. It was found that the equilibrium constant of the bonded oxidized form of the drug was 63 times bigger more important than that of the bonded reduced form. Copper forms 1 metal: 2 drug stoichiometry complex which is highly stable compared to ssDNA-drug interaction and consequently inhibited the drug biochemical damaging effects. Copper complex offered sub-nanogram determination of adriblastina in aqueous and urine media.     

Real-time measurements of noradrenaline release in periphery and central nervous system

Noradrenaline (NA) plays important hormonal and neurotransmitter roles in the periphery and central nervous system, respectively. The cells that produce and release NA, namely, adrenal chromaffin cells (ACCs), sympathetic postganglionic neurones and central neurones, show both commonalities as well as profound differences in morphology, physiological function and characteristics of NA secretion. In order to address disorders which have been associated with the dysregulation of NA release, such as essential hypertension, a better understanding of the molecular mechanisms governing and modulating NA release in neurones is urgently required. Due to profound technical challenges, the molecular basis of NA release has been investigated much more thoroughly in ACCs than in neurones. This review discusses suitable approaches for detecting NA secretion in periphery as well as brain tissues. Membrane capacitance and high-resolution electrochemical measurements have proven particularly useful when combined with fluorescence microscopy. ACCs and peripheral and central NAergic neurones are compared regarding their vesicle morphologies, as well as possible locations of release sites, and the trajectory of secreted NA. Further, current views on the properties of single vesicle release events, including proposed release probabilities in these cell types, are presented.     

Interleukin 1alpha alters hippocampal serotonin and norepinephrine release during open-field behavior in Sprague-Dawley animals: differences from the Fawn-Hooded animal model of depression

Detection of two biogenic amine neurotransmitters, serotonin (5-HT) and norepinephrine (NE) within the CA1 region of the hippocampus (HPC) of behaving male laboratory animals (Rattus norvegicus), was performed with miniature carbon sensors (BRODERICK PROBES) and in vivo semidifferential microvoltammetry after acute administration of the soluble immune factor, human recombinant, interleukin (IL) 1alpha (10 and 100 ng/kg i.p.). Two animal models were compared, i.e., (a) the Sprague-Dawley (SD) model, a strain neither biochemically nor immune-challenged and (b) the Fawn-Hooded (FH) model, a biochemically (5-HT-deficient) and immune-challenged animal. Open-field behaviors, locomotion (ambulations) and stereotypy (fine movements of sniffing and grooming) were monitored with infrared photobeams while 5-HT and NE were selectively and separately detected within seconds in real time. Subchronic studies were performed in the same animals 24 h later at which time no further drug was administered. Results from acute treatment studies showed that IL-1alpha altered HPC monoamines and behavior viz-a-viz habituation values (baseline) in the SD strain differently from those in the FH strain as follows: (1) although 5-HT release was significantly increased within CA1 region of HPC in both SD and FH strains (P<.0001), the extent of the HPC 5-HT increase in the 5-HT-deficient FH strain was significantly less than that of the SD strain at both doses (P<.0001). The subchronic studies showed that 5-HT release within the HPC in the SD strain significantly increased (135%) over drug treatment values (P<.001), whereas HPC 5-HT release in the FH strain remained the same as that seen in the acute drug treatment studies; the difference between strains for the subchronic study was also statistically significant (P<.01). (2) IL-1alpha significantly decreased HPC NE release in the SD strain (P<.0004) while IL-1alpha decreased HPC NE release in the FH strain only at the 10-ng/kg dose (P<.001); at the 100-ng/kg dose in the FH strain, NE rebounded towards baseline and increased 15% above baseline reaching statistical significance (P<.05). Subchronic studies in the SD strain showed a further decreased NE signal to 38% below baseline (P<.0001), whereas subchronic studies in the FH strain showed a significant increase in NE release (P<.02). The difference between strains in the subchronic NE studies was significant (P<.001). (3) Ambulations were increased after IL-1alpha administration in acute studies in both the SD and the FH strains, but the increase did not reach statistical significance, whereas in the subchronic studies, both strains exhibited significant increases as revealed by post hoc analyses (P<.05). There was a statistically significant difference between strains in acute studies (P<.001), whereas no significant differences between models were seen in ambulation behavior in subchronic studies. (4) Fine movements increased over baseline after IL-1alpha administration in both animal models in acute studies, however, results did not reach statistical significance, likely due to the episodic effect of IL-1alpha on movement behavior in both the SD and the FH strains. However, the SD strain showed a significant increase in fine movement behavior during the subchronic studies (P<.02). Significant differences in fine movements between animal models were not observed either acutely or in subchronic studies. In summary, the data show that immune modulation by IL-1alpha affects HPC neurochemistry and behavior in SD versus FH animal models differently and/or to different degrees. The data show that while the FH animal model is subsensitive to 5-HT agonists, 5-HT function can be stimulated. Comparison of genetically diverse animal models provides a reliable means to identify and discern cytokine-induced depressive versus stressor properties. Selective sensor technology provides a powerful tool as movement behavior is monitored and interpreted as a function of monoamine neurotransmission.     

GABAergic modulation in the substantia nigra of the striatal dopamine release and of the locomotor activity in rats exposed to helium pressure

Helium-oxygen pressure induces in rodents an increase of both locomotor and motor activity (LMA) and of the striatal dopamine release, which could result from a decrease of GABA transmission in the substantia nigra. The effects of the GABA(A) receptor agonist muscimol and of the GABA(B) receptor agonist baclofen on the striatal dopamine release were measured using differential pulse voltammetry. Behavioural studies were performed in freely moving rats using actimetry. Whatever the drug used under helium pressure, bilateral administration in the substantia nigra pars reticulata (SNr) or in the substantia nigra pars compacta (SNc) counteracted the evoked dopamine release. However, only the baclofen reduced the LMA, while the muscimol administration in the SNr, but not in the SNc, increased it. These results indicate that different subtypes of GABA receptors would be involved in the control of the DA release and in the occurrence of LMA under helium pressure.     

Absence of reinforcement with low dose intravenous ethanol self-administration in rats

Male hooded rats were implanted with intravenous cannulas and housed in operant chambers supplied with 2 levers and enclosed in sound-attenuating cubicles. In Experiment 1, seven rats received a 1.0 mg/kg infusion of ethanol for each press on the previously determined non-preferred lever. The other lever served to count "activity lever presses." An additional 7 rats served as controls and were treated identically except that each press on the non-preferred lever led to an infusion of saline, isovolumetric to the ethanol infused in the experimental subjects. The rats were tested under these conditions of continuous reinforcement for 9 days. Throughout this period, self-infusions and "activity lever presses" did not differ between the groups, suggesting that ethanol was not reinforcing at a dose of 1.0 mg/kg. These results were replicated, and extended to other low doses of ethanol in Experiment 2. Here, we employed a design where depression of either lever, under conditions of continuous reinforcement, led to the infusion of a solution. Fifteen rats were randomly assigned to one of three groups (5 rats/group). In one group, depression of the previously determined non-preferred lever led to an infusion of 16.0 mg/kg of ethanol, while depression of the other lever led to an infusion of isocaloric glucose. For the other two groups, depression of the non-preferred level led to an infusion of 4.0 and 1.0 mg/kg ethanol respectively, and depression of the other lever led to a glucose infusion. The animals were tested for 9 days, and in each case, ethanol self-infusions did not differ significantly from glucose self-infusions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Voltammetric detection of the release of 5-hydroxyindole compounds throughout the sleep-waking cycle of the rat

Summary In the present work, differential pulse voltammetry (DPV) measurements of the extracellular fraction of 5-hydroxyindole compounds were performed in rats under long-term chronic conditions. In the nucleus Raphe Dorsalis (n.RD), the voltammetric signal measured at +300 mv (peak 3) disappeared completely 70 to 90 min after injection of Clorgyline (10 mg/kg), a monoamine oxidase inhibitor type A (MAOI-A); the signal measured in such conditions is thus dependent upon extracellular 5-hydroxyindoleacetic acid (5-HIAA peak 3). Deprenyl, an MAOI type B, at the same dose, induced only a slight increase in peak 3 height; according to the fact that MAO-B is selectively located in the 5-HT neurons and since their inhibition does not decrease 5-HIAA peak 3 nor the endogenous 5-HIAA content as measured with High Performance Liquid Chromatography (HPLC), 5-HIAA measured with DPV in the extracellular fluid of untreated animals might come from 5HT released and metabolized by MAO-A outside the 5-HT neurons. In animals implanted for measurements of both voltammetric and polygraphic parameters, the 5-HIAA peak 3 measured mainly in the anterior and ventral part of the n.RD exhibited large increases in its height during slow-wave sleep (SWS: +39%) and paradoxical sleep (PS=+71%) as compared to the waking state (W=100%); these variations could reflect the dendritic release of 5-HT. In the Caudate nucleus (n.Cd) the same voltammetric signal presented reverse fluctuations, i.e. an increase during W and a decrease during SWS and PS. Intracerebroventricular administration of Corticotropin-Like Intermediate lobe Peptide (CLIP, 10 ng/2 l) induced an increase in PS duration (+51%) preceded and accompanied by an increase in the n.RD 5-HIAA peak 3 height (+50%).     

L—半胱氨酸在金电极上的电化学行为

目的 研究L－半胱氨酸在金电极上的电化学行为,为测定L－半膛氨酸的物理化学及参数及定性、定量分析提供依据。方法 在磷酸盐缓冲体系中以循环伏安法对L－半胱氨酸进行电化学表征,探讨其在金电极上的电化学反应机理,以及介质浓度、pH值等各种因素对峰电流的影响。结果 在pH值小于7时,除在0．96V处有一氧化峰外,在－0．75V左右发现一新的还原峰,氧化峰和还原峰峰电流的大小与本体溶液中半膛 氨酸的浓度成正比,且峰电流与扫速的平方根成正比。结论 0．96V处的氧化峰和－0．75V处的还原峰具有一定的相关性,L－半胱氨酸在金 电极上的吸附及其电化学行为是受扩散控制的。     

Cation effects on the voltammetric behavior of α-Keggin-type [SiMo12O40]4− and [PMo12O40]3− complexes in CH3COCH3 and CH3CN

The effect of small cations such as H⁺, Li⁺ and Na⁺ on the voltammetric behavior of α-Keggin-type [SiMo₁₂O₄₀]^4? and [PMo₁₂O₄₀]^3? complexes was investigated in CH₃COCH₃ and CH₃CN. For the [SiMo₁₂O₄₀]^4? complex, the presence of Li⁺ or Na⁺ caused the one-electron waves to be converted into a two-electron wave at ca. 0.3 V more positive than the first one-electron wave. In the presence of Li⁺ or Na⁺, the [PMo₁₂O₄₀]^3? complex underwent a two-electron reduction at the same potential as the original first one-electron wave in CH₃COCH₃, whereas it exhibited only successive one-electron waves in CH₃CN. The addition of a trace amount of H⁺ produced new two-electron waves at more positive potentials. These findings give a clue to the understanding of the reactivity of polyoxometalates as redox catalysts.     

Decrease of extracellular catecholamine content in the vicinity of cortical penicillin-induced epileptogenic focus: voltammetric study in the rat

Differential pulse voltammetry with carbonfibre microelectrodes was used in chloralhydrate-anaesthetized rats to test the influence of the penicillin-G-Na (PNC)-induced (topical application, approximately 2000 IU) epileptic activity on the catecholamine content (catechol-oxidative current, CA.OC) in the parietal cortex. In the experimental group (n = 4) after PNC a nonlinear CA.OC lowering was observed; this decrease during the first 10 min was faster than in the control group (n = 4). Significantly different values were observed from the 4th min after application. The best fit for this experimental curve gave the logarithmic function (f(t) = a + b.ln(t), a =105.8, b= -10.6) with regression coefficient r = 0.98. From the 12th min after PNC application until the end of the experiments (54th min) CA.OC values ranged from 78% to 84% of the control group.