Abstract
The isolation of a soluble brain fraction which behaves as an endogenous ouabain-like substance, termed endobain E, has been described. Endobain E contains two Na+, K+-ATPase inhibitors, one of them identical to ascorbic acid. Neurotransmitter release in the presence of endobain E and ascorbic acid was studied in non-depolarizing (0 mM KCl) and depolarizing (40 mM KCl) conditions. Synaptosomes were isolated from cerebral cortex of male Wistar rats by differential centrifugation and Percoll gradient. Synaptosomes were preincubated in HEPES-saline buffer with 1 mM d-[3H]aspartate (15 min at 37°C), centrifuged, washed, incubated in the presence of additions (60 s at 37°C) and spun down; radioactivity in the supernatants was quantified. In the presence of 0.5–5.0 mM ascorbic acid, d-[3H]aspartate release was roughly 135–215% or 110–150%, with or without 40 mM KCl, respectively. The endogenous Na+, K+-ATPase inhibitor endobain E dose-dependently increased neurotransmitter release, with values even higher in the presence of KCl, reaching 11-times control values. In the absence of KCl, addition of 0.5–10.0 mM commercial ouabain enhanced roughly 100% d-[3H]aspartate release; with 40 mM KCl a trend to increase was recorded with the lowest ouabain concentrations to achieve statistically significant difference vs. KCl above 4 mM ouabain. Experiments were performed in the presence of glutamate receptor antagonists. It was observed that MPEP (selective for mGluR5 subtype), failed to decrease endobain E response but reduced 50–60% ouabain effect; LY-367385 (selective for mGluR1 subtype) and dizocilpine (for ionotropic NMDA glutamate receptor) did not reduce endobain E or ouabain effects. These findings lead to suggest that endobain E effect on release is independent of metabotropic or ionotropic glutamate receptors, whereas that of ouabain involves mGluR5 but not mGluR1 receptor subtype. Assays performed at different temperatures indicated that in endobain E effect both exocytosis and transporter reversion are involved. It is concluded that endobain E and ascorbic acid, one of its components, due to their ability to inhibit Na+, K+-ATPase, may well modulate neurotransmitter release at synapses.
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R. D. O’Neill R. A. Grunewald M. Fillenz W. J. Albery (1983) ArticleTitleThe effect of unilateral cortical lesions on the circadian changes in rat striatal ascorbate and homovanillic acid levels measured in vivo using voltammetry Neurosci. Lett 42 105–110 Occurrence Handle1:CAS:528:DyaL2cXksF2itA%3D%3D Occurrence Handle6657142
E. A. Kiyatkin G. V. Rebec (1998) ArticleTitleAscorbate modulates glutamate-induced excitations of striatal neurons Brain Res 812 14–22 Occurrence Handle1:CAS:528:DyaK1cXntVyiu7s%3D Occurrence Handle9813223
R. Dingledine C. J. McBain (1999) Glutamate and aspartate G. J. Siegel B. W. Agranoff R. W. Albers S. K. Fisher M. D. Uhler (Eds) Basic Neurochemistry EditionNumber6 Lippincott-Raven Press Philadelphia
J. Cammack B. Ghasemzadeh R. N. Adams (1991) ArticleTitleThe pharmacological profile of glutamate-evoked ascorbic acid efflux measured by in vivo electrochemistry Brain Res 565 17–22 Occurrence Handle1:CAS:528:DyaK38XjsV2rsw%3D%3D Occurrence Handle1685348
J. Cammack B. Ghasemzadeh R. N. Adams (1992) ArticleTitleElectrochemical monitoring of brain ascorbic acid changes associated with hypoxia, spreading depression, and seizure activity Neurochem. Res 17 23–27 Occurrence Handle1:CAS:528:DyaK38XhtVeks7c%3D Occurrence Handle1347161
R. A. Grunewald (1993) ArticleTitleAscorbic acid in the brain Brain Res. Brain Res. Rev 18 123–133 Occurrence Handle1:STN:280:ByyB383ovFA%3D Occurrence Handle8467348
M. Miele M. G. Boutelle M. Fillenz (1994) ArticleTitleThe physiologically induced release of ascorbate in rat brain is dependent on impulse traffic, calcium influx and glutamate uptake Neuroscience 62 87–91 Occurrence Handle1:CAS:528:DyaK2MXot1yjsQ%3D%3D Occurrence Handle7816214
G. Rodríguez de Lores Arnaiz Particle M. Antonelli de Gómez de Lima (1986) ArticleTitlePartial characterization of an endogenous factor which modulates the effect of catecholamines on synaptosomal Na+, K+-ATPase Neurochem. Res 11 933–947 Occurrence Handle3018605
G. Rodríguez de Lores Arnaiz Particle (1992) ArticleTitleIn search of synaptosomal Na+, K+-ATPase regulators Mol. Neurobiol 6 359–375 Occurrence Handle1363048
G. Rodríguez de Lores Arnaiz Particle (1993) ArticleTitleAn endogenous factor which interacts with synaptosomal membrane Na+, K+-ATPase activation by K+ Neurochem. Res 18 655–661 Occurrence Handle8389989
G. Rodríguez de Lores Arnaiz Particle C. Peña (1995) ArticleTitleCharacterization of synaptosomal membrane Na+, K+-ATPase inhibitors Neurochem. Int 27 319–327 Occurrence Handle8845732
C. Peña G. Rodríguez de Lores Arnaiz Particle (1997) ArticleTitleDifferential properties between an endogenous brain Na+, K+-ATPase inhibitor and ouabain Neurochem. Res 22 379–383 Occurrence Handle9130247
G. Rodríguez de Lores Arnaiz Particle A. Reinés T. Herbin C. Peña (1998) ArticleTitleNa+, K+-ATPase interaction with a brain endogenous inhibitor (endobain E) Neurochem. Int 33 425–433 Occurrence Handle9874093
M. Vatta C. Peña B. Fernández G. Rodríguez de Lores Arnaiz Particle (1999) ArticleTitleA brain Na+, K+-ATPase inhibitor (endobain E) enhances norepinephrine release in rat hypothalamus Neuroscience 90 573–579 Occurrence Handle1:CAS:528:DyaK1MXitFGju7c%3D Occurrence Handle10215160
M. A Calviño C. Peña G. Rodríguez de Lores Arnaiz Particle (2001) ArticleTitleAn endogenous Na+, K+-ATPase inhibitor enhances phosphoinositide hydrolysis in neonatal but not in adult rat brain cortex Neurochem. Res 26 1253–1259 Occurrence Handle11874208
G. Rodríguez de Lores Arnaiz Particle (2000) ArticleTitleHow many endobains are there? Neurochem. Res 25 1421–1430 Occurrence Handle11059812
M. A Calviño C. Peña G. Rodríguez de Lores Arnaiz Particle (2002) ArticleTitleMetabotropic glutamate receptor involvement in phosphoinositide hydrolysis stimulation by an endogenous Na+, K+-ATPase inhibitor and ouabain in neonatal rat brain Dev. Brain Res 138 167–175
G. Rodríguez de Lores Arnaiz Particle T. Herbin C. Peña (2003) ArticleTitleA comparative study between a brain Na+, K+-ATPase inhibitor (endobain E) and ascorbic acid Neurochem. Res 28 903–910 Occurrence Handle12718444
A. Nagy A. V. Delgado-Escueta (1984) ArticleTitleRapid preparation of synaptosomes from mammalian brain using nontoxic isoosmotic gradient material (Percoll) J. Neurochem 43 1114–1123 Occurrence Handle1:CAS:528:DyaL2cXlvVOgtr4%3D Occurrence Handle6088694
P. R. Dunkley J. W. Heath S. M. Harrison P. E. Jarvie P. J. Glenfield J. A. P. Rostas (1988) ArticleTitleA rapid percoll gradient procedure for isolation of synaptosomes directly from an S1 fraction: homogeneity and morphology of subcellular fractions Brain Res 441 59–71 Occurrence Handle1:CAS:528:DyaL1cXhsVKmt7k%3D Occurrence Handle2834006
M. A. Lynch K. L. Voss (1990) ArticleTitleArachidonic acid increases inositol phospholipid metabolism and glutamate release in synaptosomes prepared from hippocampal tissue J. Neurochem 55 215–221 Occurrence Handle1:CAS:528:DyaK3cXksFejs74%3D Occurrence Handle1972388
O. H. Lowry N. J. Rosebrough A. L. Farr R. J. Randall (1951) ArticleTitleProtein measurement with the Folin phenol reagent J. Biol. Chem 193 265–275 Occurrence Handle1:CAS:528:DyaG38XhsVyrsw%3D%3D Occurrence Handle14907713
A. M. Palmer C. T. Reiter (1994) ArticleTitleComparison of the superfused efflux of preaccumulated D-[3H]aspartate and endogenous L-aspartate and L-glutamate from rat cerebrocortical minislices Neurochem. Int 25 441–450 Occurrence Handle1:CAS:528:DyaK2MXit1yjurw%3D Occurrence Handle7849572
A. C. Rego M. S. Santos C. R. Oliveira (1996) ArticleTitleOxidative stress, hypoxia, and ischemia-like conditions increase the release of endogenous amino acids by distinct mechanisms in cultured retinal cells J. Neurochem 66 2506–2516 Occurrence Handle1:CAS:528:DyaK28XjtFCku70%3D Occurrence Handle8632176
M. Fillenz (1995) ArticleTitlePhysiological release of excitatory amino acids Behav. Brain Res 71 51–67 Occurrence Handle1:CAS:528:DyaK28XhtFKlsbk%3D Occurrence Handle8747174
M. Reid N. Toms J. Bedingfield P. Roberts (1999) ArticleTitleGroup I mGlu receptors potentiate synaptosomal [3H]glutamate release independently of exogenously applied arachidonic acid Neuropharmacology 38 477–485 Occurrence Handle1:CAS:528:DyaK1MXitlGks78%3D Occurrence Handle10221751
L. S Thomas D. E. Jane J. R. Harris M. J. Croucher (2000) ArticleTitleMetabotropic glutamate autoreceptors of the mGlu5 subtype positively modulate neuronal glutamate release in the rat forebrain in vitro Neuropharmacology 39 1554–1566 Occurrence Handle1:CAS:528:DC%2BD3cXjvFGqtbw%3D Occurrence Handle10854900
E. S. Vizi (1978) ArticleTitleNa+, K+-activated adenosine triphosphatase as a trigger in transmitter release Neuroscience 3 367–384 Occurrence Handle1:CAS:528:DyaE1cXlt1Knt7c%3D Occurrence Handle28496
E. S. Vizi (1998) ArticleTitleDifferent temperature dependence of carrier-mediated (cytoplasmic) and stimulus-evoked (exocytotic) release of transmitter: a simple method to separate the two types of release Neurochem. Int 33 359–366 Occurrence Handle1:CAS:528:DyaK1cXnsFelt78%3D Occurrence Handle9840227
Z. Gerevich L. Tretter V. Adam-Vizi M. Barayani J. P. Kiss T. Zelles E. S. Vizi (2001) ArticleTitleAnalysis of high intracellular [Na+]-induced release of [3H]noradrenaline in rat hippocampal slices Neuroscience 104 761–768 Occurrence Handle1:CAS:528:DC%2BD3MXkvVegs7o%3D Occurrence Handle11440807
E. S. Vizi B. Sperlágh (1999) ArticleTitleSeparation of carrier mediated and vesicular release of GABA from rat brain slices Neurochem. Int 34 407–413 Occurrence Handle1:CAS:528:DyaK1MXktFantLo%3D Occurrence Handle10397369
J. Smythies (1999) ArticleTitleThe neurochemical basis of learning and neurocomputation: the redox theory Behav. Brain Res 99 1–6 Occurrence Handle1:CAS:528:DyaK1MXnvVWhsg%3D%3D Occurrence Handle10512566
J. Smythies (1999) ArticleTitleRedox mechanisms at the glutamate synapse and their significance: a review Eur. J. Pharmacol 370 1–7 Occurrence Handle1:CAS:528:DyaK1MXis1aktL0%3D Occurrence Handle10323273
W. L. Caudill J. C. Bigelow R. M. Wightman (1985) ArticleTitleComparison of release of endogenous dopamine and gamma-aminobutyric acid from rat caudate synaptosomes Neurochem. Res 10 319–331 Occurrence Handle1:CAS:528:DyaL2MXhvFymsLw%3D Occurrence Handle4000390
V. Castagne M. Rougemont M. Cuenod K. Q. Do (2004) ArticleTitleLow brain glutathione and ascorbic acid associated with dopamine uptake inhibition during rat’s development induce long-term cognitive deficit: relevance to schizophrenia Neurobiol. Dis 15 93–105 Occurrence Handle1:CAS:528:DC%2BD2cXntVOhsQ%3D%3D Occurrence Handle14751774
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Bersier, M.G., Miksztowicz, V., Peña, C. et al. Modulation of Aspartate Release by Ascorbic Acid and Endobain E, an Endogenous Na+, K+-ATPase Inhibitor. Neurochem Res 30, 479–486 (2005). https://doi.org/10.1007/s11064-005-2684-2
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DOI: https://doi.org/10.1007/s11064-005-2684-2