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E Schoffeniels

Publications and source records attributed to E Schoffeniels.

At least 19 recordsLinked to original sources
PubMed1996

Paradoxical sleep deprivation increases the content of glutamate and glutamine in rat cerebral cortex.

We investigated the influence of the sleep/waking cycle, the effects of paradoxical sleep deprivation (PSD) and of the vigilance-promoting drug modafinil on the amino acid contents of rat brain cortex. No significant nycthemeral variations in amino acid levels could be detected. PSD (12-24 hours), using the water tank method, significantly increased the levels of glutamate and glutamine. The increase was still observed after the sleep rebound period. gamma-Aminobutyric acid (GABA) levels did not change significantly during the instrumental sleep deprivation but increased during the rebound period. Control experiments indicate that the increase in glutamate and glutamine levels is due to PSD rather than to the stress associated with the experimental procedure. The increase in glutamate content cannot arise only from transamination reactions, because the levels of other amino acids (such as aspartate) did not decrease. Modafinil treatment did not significantly modify the brain cortex content of any of the amino acids tested.

Animals↗
PubMed1994

[Comparative biochemistry of the triune brain].

The contribution of the phylogenesis to the study of the functional organisation of brain constitutes an essential component in the integration of the current data regarding the knowledge of the central nervous system. The three evolutive structures evidenced by McLean allowed manifestation of a biological diversity particularly expressed in the neommamalian brain. Studying this biological diversity constitutes one of the field of comparative biochemistry. In this article, the authors attempt to characterize evolution through the study of GABA-receptors.

Brain Chemistry↗
PubMed1993

Thiamine triphosphate activates an anion channel of large unit conductance in neuroblastoma cells.

In neuroblastoma cells, the intracellular thiamine triphosphate (TTP) concentration was found to be about 0.5 microM, which is several times above the amount of cultured neurons or glial cells. In inside-out patches, addition of TTP (1 or 10 microM) to the bath activated an anion channel of large unit conductance (350-400 pS) in symmetrical 150 mM NaCl solution. The activation occurred after a delay of about 4 min and was not reversed when TTP was washed out. A possible explanation is that the channel has been irreversibly phosphorylated by TTP. The channel open probability (Po) shows a bell-shaped behavior as a function of pipette potential (Vp). Po is maximal for -25 mV < Vp < 10 mV and steeply decreases outside this potential range. From reversal potentials, permeability ratios of PCl/PNa = 20 and PCl/Pgluconate = 3 were estimated. ATP (5 mM) at the cytoplasmic side of the channel decreased the mean single channel conductance by about 50%, but thiamine derivatives did not affect unit conductance; 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (0.1 mM) increased the flickering of the channel between the open and closed state, finally leading to its closure. Addition of oxythiamine (1 mM), a thiamine antimetabolite, to the pipette filling solution potentiates the time-dependent inactivation of the channel at Vp = -20 mV but had the opposite effect at +30 mV. This finding corresponds to a shift of Po towards more negative resting membrane potentials. These observations agree with our previous results showing a modulation of chloride permeability by thiamine derivatives in membrane vesicles from rat brain.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Thiamin and derivatives as modulators of rat brain chloride channels.

Several membrane fractions were prepared from rat brain by differential and sucrose density gradient centrifugation. Most fractions took up 36Cl- rapidly at a rate linear with time during the first 30-60 s, then the rate progressively slowed down. The lowest rate of uptake was found in the mitochondrial fraction. Oxythiamin partially inhibited 36Cl- uptake in all fractions. In P2 (crude synaptosomal fraction), oxythiamin decreased the initial rate of uptake by 32%, the apparent Ki being 1.5 mM. Thiamin and amprolium were less effective as inhibitors. 4,4'-Diisothiocyanostilbene-2,2'-disulfonic acid (0.1-1 mM) inhibited 36Cl- uptake by 40-50%. In the presence of this compound at a concentration > or = 5 x 10(-4) M, oxythiamin became ineffective. 36Cl- uptake was increased by GABA (0.1 mM) and this effect was antagonized by picrotoxin as expected, but not by oxythiamin. The rate of 36Cl- uptake did not appreciably depend on the external chloride concentration and was unaffected by bumetanide or by replacement of external Na+ by choline. Taken together, these data suggest that the oxythiamin-sensitive 36Cl- influx is essentially diffusional and is not related to the GABA receptor or the Na:K:2Cl co-transport. Partial replacement of external Na+ by K+ or treatment with 0.1 mM veratridine (which should both result in membrane depolarization) increased 36Cl- uptake by 50 and 30% respectively; the inhibitory effect of oxythiamin was enhanced to the same proportion.(ABSTRACT TRUNCATED AT 250 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Metabolism of thiamine triphosphate in rat brain: correlation with chloride permeability.

Our results show that a net synthesis of thiamine triphosphate (TTP) can be demonstrated in vitro using rat brain extracts. The total homogenate was preincubated with thiamine or its diphosphate derivative (TDP), centrifuged, and washed twice. With TDP (1 mM) as substrate, a 10-fold increase in TTP content was observed in this fraction (nuclear fraction, membrane vesicles). A smaller, but significant, increase was observed in the P2 fraction (mitochondrial/synaptosomal fraction). In view of the low TTP content of our fractions, it was carefully assessed that authentic TTP was being formed. Incorporation of radioactivity from [beta-32P]TDP and [gamma-32P]ATP in TTP suggests that these two compounds are its precursors. Furthermore, TTP synthesis was inhibited by ADP and relatively low concentrations of Zn2+. These results suggest that TTP synthesis is catalyzed by an ATP:TDP transphosphorylase rather than by the cytoplasmic adenylate kinase that may be present in the vesicles. After osmotic lysis of the vesicles at alkaline pH, TTP was recovered in protein-bound form. Concomitantly, a soluble thiamine triphosphatase, with alkaline pH optimum, was also released from the vesicles. No net synthesis could be obtained in the cytosolic fraction or in detergent-solubilized systems. Like TTP synthesis, chloride permeability of the vesicles was increased when the homogenate had been incubated with thiamine and particularly with TDP. Our results suggest a regulatory role of TTP on chloride permeability, but the target remains to be characterized.

Animals↗
PubMed1991

Solubilization of thiamine triphosphatase from the electric organ of Electrophorus electricus.

The membrane-associated, anion-regulated thiamine triphosphatase from Electrophorus electricus electric organ can be solubilized by various neutral detergents. Polyoxyethylene ethers are the most effective. Anionic detergents readily inactivate the enzyme. A 6.4-fold increase in specific activity is obtained by successive treatment of crude membranes with octanoyl-N-methylglucamide, which solubilized other proteins, and Lubrol-PX with releases 60% of the thiamine triphosphatase (TTPase) activity. Solubilization by Lubrol-PX rapidly modifies kinetic parameters. The Km, Vmax and pH optimum are decreased. However, the solubilized TTPase may be kept at 0 degrees C for many hours without further change in specific activity. At 35 degrees C, the half-life is still 83 min at pH 5.0, but denaturation becomes rapid at pH greater than or equal to 7. Solubilization modifies anion effects on TTPase activity. The activating effect of nitrate is nearly lost, while inhibition by sulfate is no longer time-dependent.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1991

Determination of thiamin and its phosphate esters in cultured neurons and astrocytes using an ion-pair reversed-phase high-performance liquid chromatographic method.

A sensitive method, based on fluorescence detection, for the determination of thiamin derivatives after precolumn derivatization is described. The separation is achieved on a PRP-1 column using ion-pair reversed-phase HPLC. This method is especially well adapted to the detection of thiamin triphosphate in complex mixtures such as tissue extracts. The detection limit for TTP is 50 fmol. The contents of thiamin derivatives were determined in primary cultures of rat cerebellar granule neurons and cerebral astrocytes. The amount of TTP is about five times higher in neurons than in astrocytes. Thus in rat brain TTP seems to be essentially associated with neurons and the intracellular concentration is estimated to be about 0.2 microM. Our results suggest the existence, in nerve cells, of specific regulatory mechanisms not related to the blood-brain barrier and responsible for the maintenance of thiamin homeostasis in brain.

Animals↗
PubMed1990

Regulation of ion uptake in membrane vesicles from rat brain by thiamine compounds.

We examined the effects of thiamine derivatives on ion uptake in rat brain membrane vesicles. Thiamine triphosphate (1 mM) and pyrithiamine (0.1 mM) increase chloride uptake. Preincubation of crude homogenate with thiamine or pyrithiamine increases chloride uptake while oxythiamine has the reverse effect. Thiamine and oxythiamine also affect 22Na+ and 86Rb+ uptake in the same way as for 36Cl- but to a lesser extent. Thiamine-dependent 36Cl- uptake is activated by sodium bicarbonate (10 mM) and partially inhibited by bumetanide (0.1 mM) and 2,4-dinitrophenol (0.1 mM). Preincubation with thiamine increases the thiamine triphosphate content of the vesicles. The hypothesis that TTP is the activator of a particular chloride uptake mechanism is discussed.

Animals↗
PubMed1990

Molecular aspects of human brain sodium channel.

The sodium channel content of human brain was measured by tritiated tetrodotoxin specific binding. After solubilization, the sodium channel was submitted to chromatography on diethylaminoethyl(cellulose) Sephadex, hydroxylapatite and wheat germ agglutinin sepharose. An increase of tritiated tetrodotoxin binding specific activity was subsequently observed. Eluted sodium channels from wheat germ agglutinin sepharose were overlaid on a sucrose gradient. Electrophoretical analysis of the material obtained after the sedimentation step revealed two co-purified peptides, alpha (Mr = 275,000 mol. wt) and beta (Mr = 30,000-36,000 mol. wt.). Alpha showed an exceptionally high free electrophoretic mobility, which is a common feature for all sodium channels previously described. However, the high denaturation rate of the solubilized tetrodotoxin receptor site 1 did not allow tetrodotoxin receptor quantification by the tritiated toxin binding in sucrose fractions. Sodium channel effective reconstitution in liposomes was demonstrated: (1) 22Na+ influx in proteoliposomes was sensitive to sodium channel-specific neurotoxins: (2) reconstituted proteins showed a cation selectivity similar to that previously described for animal sodium channels. The sodium channel preparation obtained after four chromatographic steps shows two peptides on the electrophoretic analysis. Reconstituted sodium channels displayed some physiological properties found in intact conducting membranes.

Brain↗
PubMed1989

Phosphorylated thiamine derivatives and cortical activity in the baboon Papio papio: effect of intermittent light stimulation.

The effect of intermittent light stimulation (ILS) on the distribution of thiamine derivatives in three brain areas (occipital, motor, and premotor) was compared in photosensitive and nonphotosensitive baboons. ILS induces paroxysmal discharges in the motor and premotor areas of photosensitive animals only. In baboons submitted to ILS, thiamine triphosphate (TTP) decreases in both photosensitive and nonphotosensitive animals; thiamine monophosphate (TMP) increases in photosensitive animals, which present ILS-induced paroxysmal discharges, whereas it is unaffected in nonphotosensitive animals. The variations are the most significant in the occipital (visual) cortex. A consumption of TTP may result from electrical activity induced by light stimulation in the occipital area. No correlation between ILS-induced paroxysmal activity and a decrease in TTP contents was found. However, photosensitive animals are affected differently from nonphotosensitive animals, as their content of TMP in the cerebral cortex increases on stimulation. However, as long as the exact role of thiamine compounds in relation to membrane excitability in the nervous system remains unknown, it is impossible to conclude whether the differences observed in the metabolism of thiamine compounds are the cause or the consequence of the photosensitivity in the baboon Papio papio.

Animals↗
PubMed1989

Solubilization of sodium channel from human brain.

[3H]Tetrodotoxin binds to a single class of receptor sites in homogenates of human brain with a KD of 9.1 nM at 0 degree C and a maximal binding capacity of 5.9 pmol/mg of protein. This tetrodotoxin receptor has been solubilized, and several parameters influencing the efficiency of this critical step have been studied. Treatment of brain membranes with 2% (wt/vol) Nonidet P-40 solubilizes up to 38% of the tetrodotoxin receptor sites. The duration of this solubilization step must not exceed 15 min at an optimal pH of 6.8. The binding activity is most stable when exogenous phosphatidylcholine is added to the soluble receptor with a phosphatidylcholine/detergent ratio of 1:5.

Adult↗
PubMed1989

Thiamine triphosphatase in the membranes of the main electric organ of Electrophorus electricus: substrate-enzyme interactions.

The main electric organ of Electrophorus electricus is particularly rich in thiamine triphosphate (TTP). Membrane fractions prepared from this tissue contain a thiamine triphosphatase that is strongly activated by anions and irreversibly inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), an anion transport inhibitor. Kinetic parameters of the enzyme are markedly affected by the conditions of enzyme preparation: In crude membranes, the apparent Km is 1.8 mM and the pH optimum is 6.8, but trypsin treatment of these membranes or their purification on a sucrose gradient decreases both the apparent Km (to 0.2 mM) and the pH optimum (to 5.0). Anions such as NO3- (250 mM) have the opposite effect, i.e., even in purified membranes, the pH optimum is now 7.8 and the Km is 1.1 mM; at pH 7.8, NO3- increases the Vmax 24-fold. TTP protects against inhibition by DIDS, and the KD for TTP could be estimated to be 0.25 mM, a value close to the apparent Km measured in the same purified membrane preparation. Thiamine pyrophosphate (0.1 mM) did not protect against DIDS inhibition. At lower (10(-5)-10(-6) M) substrate concentrations, Lineweaver-Burk plots of thiamine triphosphatase activity markedly deviate from linearity, with the curve being concave downward. This suggests either anticooperative binding or the existence of binding sites with different affinities for TTP. The latter possibility is supported by binding data obtained using [gamma-32P]TTP. Our data suggest the existence of a high-affinity binding site (KD of approximately 0.5 microM) for the Mg-TTP complex.(ABSTRACT TRUNCATED AT 250 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1989

[Molecular aspects of bioelectrogenesis].

The action potential is a dissipative process producing entropy and using free energy. This is well demonstrated by: 1) the evolution of the Na conductance under voltage clamping conditions, 2) the microcalorimetric measurements, 3) the analysis of heat evolution during the conductance changes. The most appropriate explanation must involve an exogenous energy source since the energy dissipated by the ionic flows or even the applied stimulus depolarization are far too small to account for the overall energy balance. Thiamine triphosphate is a likely candidate as specific operating substance. The more so, since it is specifically hydrolyzed by a triphosphatase the activity of which is modulated by various anions. It is therefore suggested that the control of the Cl-permeability, a process requiring the hydrolysis of thiamine triphosphate, is the key to our understanding of the energetics of the action potential.

Action Potentials↗