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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 262 (1969), S. 208-220 
    ISSN: 1432-1912
    Keywords: Splenic Nerves ; Storage Granules ; Noradrenaline Uptake ; ATPase ; Reserpine ; Prenylamine ; NEM ; Milznervengranula ; Noradrenalinaufnahme ; ATPase ; Reserpin ; Prenylamin ; NEM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung 1. Aus postganglionären sympathischen Rindermilznerven wurde durch Gradientenzentrifugation eine spezifische Granulafraktion (III) gewonnen, die 843±149 ng Noradrenalin/mg Protein enthielt. In dieser Fraktion war die Aktivität des mitochondrialen Enzyms Fumarase an der Grenze der Nachweisbarkeit. 2. Nach Inkubation mit 5·10−5 M Noradrenalin, 2,5 mM Mg++ and 5 mM ATP war der Amingehalt der Granula ungefähr doppelt so hoch wie nach Inkubation mit Noradrenalin allein. Dagegen beeinflußten 2,5 mM Ca++ und 5 mM ATP den Noradrenalingehalt nicht. 3. Die Milznervengranulafraktion enthält ATP-spaltende Aktivität, die durch Mg++ (K m=1,92 mM) und Ca++ (K m=1,93 mM) in gleichem Ausmaß gesteigert wird. Kombinierte Zugabe beider Ionen wirkt additiv aktivierend auf die ATP-Spaltung. Die ATPase wird in Anwesenheit von Mg++ durch Na+ und K+ nicht stimuliert und durch 10−4 M g-Strophanthin nicht gehemmt. 4. Die ATPase einer unspezifischen, noradrenalinarmen Fraktion (I) aus Milznerven war durch Ca++ stärker (K m=1,09 mM) als durch Mg++ (K m=3,00 mM) aktivierbar und hatte eine größere maximale Reaktionsgeschwindigkeit als die ATPase der Fraktion III. 5. 6·10−5 M Reserpin bzw. Prenylamin hemmten die Noradrenalinaufnahme und die ATP-Spaltung. 1,8·10−4 M N-äthylmaleimid hatte auf beide Prozesse keinen Einfluß. 6. Eine kausale Beziehung zwischen Amintransport und ATPase-Aktivität in Milznervengranula wird diskutiert.
    Notes: Summary 1. Homogenates of splenic sympathetic nerves from cattle were used to prepare, by density gradient centrifugation, a specific granule fraction (III), which contains 843±149 ng noradrenaline/mg protein. This fraction did not show any appreciable fumarase activity. 2. Incubation of the granule fraction in the presence of noradrenaline (5×10−5 M) resulted in a nearly doubled noradrenaline content if Mg++ (2.5 mM) and ATP (5 mM) were added. The addition of Ca++ (2.5 mM) and ATP (5 mM), on the contrary, did not increase the noradrenaline content of the granules. 3. The ATPase activity of nerve granules can be stimulated to the same degree by Mg++ (K m=1.92) and Ca++ (K m=1.93 mM). A combination of both cations additively activated the hydrolysis of ATP. In the presence of Mg++ this ATPase is neither stimulated by Na+ (30 mM) and K+ (20 mM) nor inhibited by ouabain (10−4 M). 4. The ATPase activity of an unspecific fraction (I) obtained from splenic nerve homogenates could be stimulated to a greater extent by Ca++ (K m=1.09 mM) than by Mg++ (K m=3.00 mM). The ATPase of fraction I showed a maximal velocity of reaction higher than that that of fraction III. 5. The uptake of noradrenaline and the hydrolysis of ATP were inhibited by 6×10−5 M reserpine or prenylamine. N-ethylmaleimide (1.8×10−4 M) did not influence either reactions. 6. A causal relationship between amine transport and ATPase activity in the noradrenaline storing nerve granules is discussed.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 287 (1975), S. 181-190 
    ISSN: 1432-1912
    Keywords: Caudate Nucleus ; Dopamine ; Ouabain ; Reserpine ; Prenylamine ; Uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Homogenates of the caudate nucleus of the pig were submitted to differential centrifugation. The 20 000 g and 80 000 g fractions were isolated and a part of them was osmotically shocked. The highest dopamine content per mg protein was found in the intact 80 000 g fraction. Incubation experiments with the intact and the osmotically shocked fractions at 25°C revealed that the particles of the intact 20 000 g fraction took up dopamine; the influx of the amine was not enhanced by addition of ATP and magnesium to the incubation medium. On the other hand after osmotic shock the uptake of dopamine into the particles of this fraction was greatly enhanced by addition of ATP and magnesium. The uptake of dopamine into the particles of both intact and osmotically shocked 80 000 g fractions was likewise enhanced by ATP and magnesium. The uptake in all fractions was not influenced by ouabain. The influx of dopamine into the particles of the intact 80 000 g fraction was competitively inhibited by reserpine (K i 0.96×10−8 M) and prenylamine (K i 1.74×10−8 M). It is concluded that the intact 20,000 g fraction contains intact synaptosomes; the uptake of dopamine is independent of the presence of ATP and magnesium. The shocked 20 000 g fraction and the 80 000 g fractions contain synaptic vesicles; the uptake of dopamine into these vesicles is enhanced by ATP and magnesium.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 300 (1977), S. 25-30 
    ISSN: 1432-1912
    Keywords: Synaptic vesicles ; Uptake ; Dopamine ; Tyramine ; Reserpine ; Filtration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Synaptic vesicles isolated from the caudate nucleus of the pig were preincubated with reserpine, serotonin or tyramine at concentrations which caused a 90% inhibition of the ATP-Mg2+-dependent uptake of 14C-dopamine. The preincubated vesicles were sedimented by centrifugation and resuspended in drug-free buffer. The vesicles were incubated with 14C-dopamine in the presence or in the absence of ATP-Mg2+; at the end of the incubation period they were separated from the incubation medium by centrifugation. The inhibitory effect of reserpine on the ATP-Mg2+-dependent uptake of 14C-dopamine was slightly diminished, that of serotonin reduced while the inhibitory effect of tyramine was abolished. Hence, tyramine seems to be more easily removable than the other substances. In another series of experiments the vesicles were incubated with 3H-tyramine and immediately separated by filtration through membrane filters. Addition of ATP-Mg2+ enhanced the uptake of 3H-tyramine. The Km of the ATP-Mg2+-dependent uptake of 3H-tyramine was 1.4×10−7 M, the Vmax 29.2 pmoles/mg protein/min. At 0°C the uptake of 3H-tyramine in the absence of ATP-Mg2+ was reduced, that in the presence of ATP-Mg2+ abolished. Incubation of the vesicles with 3H-dopamine revealed K m and V max values similar to those previously found when the vesicles were isolated from the incubation medium by centrifugation. Incubation in the presence of reserpine inhibited the ATP-Mg2+-dependent uptake of 3H-tyramine (IC50 1.4×10−8 M) and 3H-dopamine (IC50 4.1×10−8 M). The results demonstrate that tyramine is taken up into the vesicles by a process which is dependent on ATP-Mg2+ and temperature. Failure of previous attempts to prove an ATP-Mg2+-dependent uptake of tyramine when the vesicles were separated by centrifugation seems to be due to release of the accumulated tyramine during the centrifugation procedure.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 282 (1974), S. 1-8 
    ISSN: 1432-1912
    Keywords: Hypothalamus ; Release ; Noradrenaline ; Dopamine ; Amphetamine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The posterior hypothalamus of cats immobilized with gallamine was superfused through a push-pull cannula with artificial cerebrospinal fluid. Addition of3H-tyrosine into the superfusing fluid led to synthesis of3H-catecholamines which were released spontaneously. Separation of the3H-catecholamines by column chromatography or their acetylation and separation by paper chromatography revealed that both3H-noradrenaline and3H-dopamine were released. In most experiments3H-noradrenaline represented about 10 to 25% of total3H-catecholamines. Superfusion of the hypothalamus with amphetamine (1×10−5 M) enhanced the release of total3H-catecholamines, the release of3H-noradrenaline being relatively more enhanced than that of3H-dopamine. Determination of the readioactive compounds in the hypothalamus at the end of the experiments showed that total3H-catecholamines represented 3% of3H-tyrosine. About 15% of the total3H-catecholamines were due to3H-noradrenaline and 85% to3H-dopamine.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 347 (1993), S. 28-33 
    ISSN: 1432-1912
    Keywords: Ultradian rhythm ; Hypothalamus ; Pushpull cannula ; Dopamine ; Noradrenaline ; Adrenaline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary To investigate the patterns of catecholamine release in the brain, the hypothalamus of conscious, freely moving rats was superfused through a push-pull cannula with artificial cerebrospinal fluid and the catecholamines dopamine, noradrenaline and adrenaline were determined in the superfusate radioenzymatically. Superfusate was continuously collected in time periods of 20 min for at least 20h. Dopamine, noradrenaline and adrenaline release rates fluctuated according to an ultradian rhythm with a frequency of 1 cycle/92 min (dopamine and noradrenaline) or 99 min (adrenaline). Additionally, the three catecholamines were released according to an ultradian rhythm with the following frequencies: noradrenaline and adrenaline 1 cycle/ 12 h, dopamine 1 cycle/8 h. The release rates of dopamine and adrenaline were similar during light and dark periods, while the release rate of noradrenaline in the dark period was slightly lower than that during the light period. It is concluded that in the hypothalamus of the conscious rat the release rates of dopamine, noradrenaline and adrenaline fluctuate according to two ultradian rhythms with different frequencies.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 350 (1994), S. 339-345 
    ISSN: 1432-1912
    Keywords: Locus coeruleus ; Dopamine ; Noradrenaline ; Adrenaline ; Veratridine ; Tetrodotoxin ; Push-pull cannula ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To investigate the release of endogenous dopamine, noradrenaline and adrenaline in the locus coeruleus, this brain area was superfused with artificial cerebrospinal fluid (CSF) through push-pull cannulae and the release of catecholamines was determined in the superfusate radioenzymatically. Collection of superfusates in time periods of 10 min revealed that release rates of the three catecholamines fluctuated, thus pointing to the existence of ultradian rhythms with following mean periods (minutes per cycle): noradrenaline 52±4, dopamine 37±2, adrenaline 36±2. The rhythm frequency of noradrenaline was significantly lower than the frequencies of dopamine and adrenaline. When the locus coeruleus was superfused with neuroactive drugs, superfusates were collected in time periods of 3 min. Superfusion with tetrodotoxin (1 μmol 1−1) for 12 min elicited a prompt and sustained decrease (−70%) in the release rates of dopamine and adrenaline. The release rate of noradrenaline was also reduced, although to a lesser extent (−40%). Superfusion with veratridine (50 μmol 1−1) led to an immediate and very pronounced enhancement in the release rates of dopamine, noradrenaline and adrenaline. The veratridine-induced increase in catecholamine outflow was decreased strongly by simultaneous superfusion with tetrodotoxin. The findings suggest that the release of endogenous catecholamines in the locus coeruleus fluctuates according to ultradian rhythms. Changes in the release on superfusion with veratridine and tetrodotoxin demonstrate the neuronal origin of the three catecholamines. The observed differences in the release characteristics between noradrenaline on the one hand and dopamine and adrenaline on the other might indicate that noradrenaline is partly released from somatodendritic sites of the noradrenergic cell bodies in the locus coeruleus.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 287 (1975), S. 191-204 
    ISSN: 1432-1912
    Keywords: Synaptic Vesicles ; Dopamine ; GABA ; Serotonin ; Histamine ; ATP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Dopamine-containing vesicles were isolated from the caudate nucleus of the pig by differential centrifugation and incubated with 14C-serotonin (14C-5-HT), 14C-gamma-aminobutyric acid (14C-GABA), 14C-dopamine or 14C-histamine. At 25°C the uptake of 14C-5-HT and 14C-GABA was enhanced by ATP and magnesium. The Km for the uptake of 14C-5-HT in the presence of ATP and magnesium was 0.67×10−6 M, that of 14C-GABA 1.33×10−4 M. 14C-Histamine was also taken up into the vesicles but its uptake was not influenced by ATP and magnesim. The ATP-magnesium-dependent uptake of 14C-5-HT and 14C-GABA was abolished at 0°C. At 37°C the accumulation of 14C-GABA in the presence of ATP and magnesium reached a steady state after 20 min, while the accumulation of 14C-5-HT reached a maximum after 2.5 min of incubation and then gradually declined. Osmotic lysis of the vesicles followed by sonication abolished the enhanced uptake of 14C-GABA, 14C-5-HT and 14C-dopamine in the presence of ATP and magnesium. Determination of the endogenous contents of the vesicles revealed 14.6 pmoles per mg protein for 5-HT and 22.7 nmoles per mg protein for GABA. It is concluded that the caudate nucles preparation used in this study contains not only dopamine-storing vesicles able to take up biogenic amines but also GABA-containing vesicles which take up GABA by an ATP-magnesium-dependent process.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 339 (1989), S. 298-301 
    ISSN: 1432-1912
    Keywords: Dopamine ; Noradrenaline ; Adrenaline ; Push-pull cannula ; Rhythmic release ; Nucleus tractus solitarii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The release of endogenous dopamine, noradrenaline and adrenaline was studied in the nucleus tractus solitarii (subnucleus medialis) of anaesthetized cats. Two push-pull cannulae were inserted into the right and left nuclei tractus solitarii which were simultaneously superfused with artificial cerebrospinal fluid. The release of the neurotransmitters was determined in the superfusates which were continuously collected in 10 min or 2.5 min time periods for 6 h or 2 h, respectively. Collection of the superfusates in time periods of 10 min showed that the release rates of endogenous dopamine, noradrenaline and adrenaline varied rhythmically according to an ultradian rhythm, the time interval between 2 adjacent phases of high release rate being approximately 1 h. Some of the phases of high release rates of the three endogenous catecholamines timely coincided with each other. Collection of the perfusates in time periods of 2.5 min revealed the existence of additional oscillations with a frequency of about 1 cycle/10 min for dopamine, noradrenaline and adrenaline. The findings suggest that catecholamines are released in the nucleus tractus solitarii according to an ultradian rhythm with low (1 cycle per h) and high (1 cycle per 10 min) frequencies, which might reflect the activity of catecholaminergic neurons in this brain structure.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 258 (1967), S. 251-265 
    ISSN: 1432-1912
    Keywords: Stellate ganglion ; Noradrenaline ; Release ; Uptake ; Reserpine ; ATPase ; Ganglion stellatum ; Noradrenalin ; Freisetzung ; Aufnahme ; Reserpin ; ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung 1. Noradrenalin speichernde Granula vom Ganglion stellatum des Rindes wurden durch Differentialzentrifugieren isoliert und bei 22°C 45 min lang inkubiert. 2. Die spontane Noradrenalin-Freisetzung aus Ganglien-Granula ist etwa gleich groß wie die aus isolierten Herz- und Nerven-Granula. Sie läßt sich durch Reserpin (0,06 μMol/ml) sowie durch kleine Prenylamin-Konzentrationen (0,03 μMol/ml) hemmen, während große Prenylamin-Konzentrationen (1,2 μMol/ml) eine nahezu vollständige Noradrenalin-Verarmung der Granula verursachen. Bei der Inkubation mit ATP-Magnesium (je 3 μMol/ml) wird wahrscheinlich die Wiederaufnahme des spontan freigesetzten Noradrenalins so stark aktiviert, daß der Noradrenalin-Gehalt der Granula praktisch nicht abnimmt. 3. Isolierte Ganglien-Granula nehmen sowohl durch passive Diffusion als auch durch einen von ATP und Magnesium abhängigen Mechanismus Adrenalin auf. Die durch ATP-Magnesium bedingte Aminaufnahme läßt sich durch Reserpin hemmen. 4. Die über einem Dichtegradienten getrennten Granula stellen eine homogene, mitochondrienfreie Population dar, wie aus elektronenmikroskopischen Untersuchungen sowie aus der Bestimmung der Fumarase-Aktivität der Granula hervorgeht. Sie besitzen eine ATPase-Aktivität, die durch Reserpin und Prenylamin gehemmt wird. 5. Die Ergebnisse sprechen für eine kausale Beziehung zwischen Amin-Transport durch die Membran der sympathischen Ganglien-Granula und ihrer ATPase-Aktivität.
    Notes: Summary 1. The noradrenaline storing granules of stellate ganglia from cattle were isolated by differential centrifugation and incubated for 45 min at 22°C. 2. Noradrenaline is released spontaneously from ganglion granules to about the same extent as from heart and splenic nerve granules. Reserpine (0.06 μmoles/ml) and small concentrations of prenylamine (0.03 μmoles/ml) inhibit the spontaneous release whereas large concentrations of prenylamine (1.2 μmoles/ml) produce an almost complete depletion of noradrenaline. In the presence of ATP-Magnesium (3 μmoles/ml each) the spontaneous noradrenaline release from ganglion granules is inhibited very likely by stimulation of the reuptake mechanism for noradrenaline. 3. Adrenaline is taken up into the isolated ganglion granules by passive diffusion and by an ATP-Magnesium dependent mechanism. Only the latter one is inhibited by reserpine. 4. In order to obtain a preparation free of mitochondria ganglion granules isolated by differential centrifugation were further fractionated over a sucrose density gradient. Electron microscopic studies as well as determinations of the fumarase activity show that the obtained ganglion granules are free of mitochondria and contain practically no fumarase activity. On the other hand they do possess an ATPase activity which is inhibited by reserpine and prenylamine. 5. It is concluded that there exists a causal relationship between the transport of amines through the membrane of the ganglion granules and their ATPase activity.
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