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K Starke

Publications and source records attributed to K Starke.

At least 73 records · Page 4Linked to original sources
PubMed1996

Opposite modulation of noradrenaline and ATP release in guinea-pig vas deferens through prejunctional beta-adrenoceptors: evidence for the beta 2 subtype.

Activation of prejunctional beta-adrenoceptors has been suggested to increase the release of noradrenaline but to decrease the neural release of ATP in the guinea-pig vas deferens. Experiments were carried out to determine the subtype of beta-adrenoceptor involved. In [3H]-noradrenaline-preincubated tissues superfused with medium containing prazosin and suramin, isoprenaline (1-100 nM), salbutamol (0.01-1 microM) and terbutaline (0.1-10 microM) increased the overflow of tritium but reduced the overflow of ATP elicited by electrical stimulation (210 pulses/7 Hz). The effects of isoprenaline were blocked by the beta 2-selective antagonist 1-[2,3-(dihydro-7-methyl-1H-inden-4-yl)oxy]-3- [(1-methylethyl)amino]-2-butanol (ICI 118,551; 100 nM). In prazosin- and suramin-free medium, isoprenaline (100 nM) did not change the overflow of ATP elicited by exogenous noradrenaline (10 microM). Isoprenaline (1-100 nM), salbutamol (0.01-1 microM) and terbutaline (0.1-10 microM) reduced the initial twitch contraction elicited by electrical stimulation (210 pulses/7 Hz) in prazosin- and suramin-free medium as well as the isolated purinergic neurogenic contraction obtained by exposure to prazosin. They increased or tended to increase the secondary sustained concentration elicited by electrical stimulation in prazosin- and suramin-free medium as well as the isolated adrenergic neurogenic contraction obtained in the presence of suramin. The inhibition by isoprenaline of the isolated purinergic contraction was attenuated by ICI 118,551 (100 nM) but not by the beta 1-selective antagonist 1-[2-((3-carbamoyl- 4-hydroxy)phenoxy)ethylamino]-3-[4-(1-methyl-4- trifluoromethyl-2-imidazolyl)phenoxy]-2-propanol (CGP 20712A; 100 nM). The results confirm the opposite beta-adrenoceptor-mediated modulation of noradrenaline and neural ATP release in the guinea-pig vas deferens. They show that the prejunctional beta-adrenoceptor is of the beta 2-subtype.

Adenosine Triphosphate↗
PubMed1996

Evaluation of P2-purinoceptor antagonists at two relaxation-mediating P2-purinoceptors in guinea-pig taenia coli.

The guinea-pig taenia coli possesses two relaxation-mediating receptors for nucleotides: a prototypic P2Y-purinoceptor, which is activated by adenosine 5'-O-(2-thio-diphosphate) (ADP beta S), and a separate receptor for alpha, beta-methylene ATP (alpha,beta-MeATP). Effects of several as yet incompletely characterized P2-purinoceptor antagonists at these receptors were examined. The concentration-relaxation curve of ADP beta S was shifted to the right by reactive blue 2, suramin, 8-(3,5-dinitro-phenylenecarbonylimino)-1,3,5-naphthalenetrisulp honic acid (XAMR0721; at 1000 microM only), pyridoxalphosphate-6-azophenyl-2',5'-disulphonic acid (iso-PPADS), pyridoxal 5-phosphate, trypan blue and Evans blue (at 320 microM only). Schild plots for the antagonism of reactive blue 2, suramin, iso-PPADS and pyridoxal 5-phosphate against ADP beta S had slopes < 1. The concentration-relaxation curve of alpha,beta-MeATP was shifted to the right by reactive blue 2, suramin, XAMR0721, iso-PPADS, pyridoxal 5-phosphate and trypan blue but not by Evans blue (320 microM). Schild plots for the antagonism of suramin, XAMR0721 and iso-PPADS against alpha,beta-MeATP had slopes > 1. Only XAMR0721 differed clearly in potency against the two nucleotides: it was considerably more potent against alpha,beta-MeATP than against ADP beta S. 2-Methylthio ATP (MeSATP; 1 microM) and ATP (100 microM) were degraded by pieces of taenia coli. All antagonists except trypan blue attenuated the degradation of either or one of the two nucleotides. The selective effect of XAMR0721 against alpha,beta-MeATP confirms the existence of two relaxation-mediating P2-purinoceptors in guinea-pig taenia coli. Comparison of the apparent affinities of the antagonists for the two taenia coli receptors with affinities for the P2X-purinoceptor of the rat vas deferens shows that reactive blue 2, suramin, iso-PPADS, pyridoxal 5-phosphate and trypan blue have little selectivity for any of the three receptors. XAMR0721, which has been shown to possess relatively high affinity for the P2Y-purinoceptor in turkey erythrocytes, was very weak at the P2Y-receptor of the taenia, thus supporting the existence of pharmacologic P2Y-receptor subtypes. Evans blue, with little effect in the taenia coli but a marked effect in the rat vas defrens, is the most selective P2X- (versus P2Y-) purinoceptor antagonists presently known, although its effect on the degradation of nucleotides must be kept in mind.

Adrenergic alpha-Agonists↗
PubMed1996

P2-purinoceptors on postganglionic sympathetic neurones.

1. Postganglionic sympathetic neurones possess both excitatory and inhibitory P2-purinoceptors. 2. The mechanisms of action of excitatory P2-purinoceptors have recently been studied on cultured sympathetic neurones of the rat. The receptors mediate fast increases in intracellular Ca2+ levels and a release of noradrenaline. They are likely to belong to the neuronal types of P2X-purinoceptors and to be located on the sympathetic nerve cell bodies or their dendrites. 3. Inhibitory P2-purinoceptors have been shown to operate at sympathetic axon terminals in isolated tissues. Adenine nucleotides decreased the stimulation-evoked release of noradrenaline by activation of these receptors. The receptors are likely to belong to the group of G-protein-coupled P2Y-purinoceptors. They mediate a negative feedback in which co-transmitter ATP inhibits subsequent sympathetic transmitter release.

Action Potentials↗
PubMed1996

The presynaptic alpha-2 autoreceptors in pig brain cortex are alpha-2A.

The presynaptic alpha-2 autoreceptors in pig brain cortex were subclassified in terms of alpha-2A, alpha-2B, alpha-2C and alpha-2D to test the hypothesis that alpha-2 autoreceptors belong predominantly to the alpha-2A/D pair of orthologous alpha-2 adrenoceptors. Slices of brain cortex were preincubated with [3H]norepinephrine and then superfused and stimulated electrically. pKd values of 13 alpha-2 adrenoceptor antagonists (including the partial agonist oxymetazoline) against the alpha-2 agoinst 5-bromo-6-(2-imidazolin-2-ylamino)quinoxaline (UK 14,304) were determined. The stimulation periods used (six pulses at 100 Hz) did not lead to alpha-2 autoinhibition, as shown by the failure of all but one of the alpha-2 antagonists to increase the stimulation-evoked overflow of tritium. UK 14,304 caused a concentration-dependent decrease of the evoked overflow of tritium, with an EC50 value of 0.90 nM and a maximal inhibition of 95.2%. All antagonists shifted the concentration-inhibition curve of UK 14,304 to the right in a parallel manner. Antagonist pKd values were calculated from the shifts. The pKd values at the presynaptic alpha-2 autoreceptors in pig brain cortex correlated excellently with pKd values at previously subclassified alpha-2A sites but did not correlate significantly or correlated much less well with pKd values at alpha-2B, alpha-2C and alpha-2D sites. Also, ratios of Kd values of the antagonists at the presynaptic alpha-2 autoreceptors in pig brain cortex agreed well with ratios at previously subclassified alpha-2A sites but not with those at previously subclassified alpha-2B-D sites. It is concluded that the alpha-2 autoreceptors in pig brain cortex are alpha-2A, in accordance with the hypothesis mentioned.

Adrenergic alpha-Agonists↗
PubMed1995

Involvement of peripheral presynaptic inhibition in the reduction of sympathetic tone by moxonidine, rilmenidine and UK 14304.

We studied the possibility that presynaptic inhibition of transmitter release from postganglionic sympathetic neurons contributes to the overall reduction of sympathetic tone produced by moxonidine, rilmenidine and 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline tartrate (UK 14304). In pithed rabbits without electric stimulation, moxonidine, rilmenidine and UK 14304 caused a long-lasting, > 10 min, increase in arterial pressure. Heart rate was not changed. In pithed rabbits in which sympathetic tone was created by electric stimulation through the pithing rod (2 Hz), the same doses of moxonidine, rilmenidine and UK 14304 caused only a brief, < 10 min, blood pressure rise. Heart rate was decreased, as were the plasma concentrations of noradrenaline and adrenaline. Dose-response curves for the effects on the plasma noradrenaline concentration (stimulated pithed rabbits) were compared with previously obtained dose-response curves for depression of renal sympathetic nerve activity (conscious rabbits). For each drug, the curve describing peripheral presynaptic inhibition and the curve describing central sympathoinhibition were very similar. Both the power and the dose dependence of the peripheral inhibitory effect support its contribution to the overall decrease in sympathetic tone produced by clonidine-like drugs in intact animals. The peripheral effect in all likelihood consists in activation of presynaptic alpha 2-autoreceptors. The agreement of the dose-response curves for the peripheral and for the central effect supports the view that the central effect, like the peripheral one, is mediated through alpha 2-adrenoceptors.

Adrenergic alpha-2 Receptor Agonists↗
PubMed1995

Involvement of alpha 2-adrenoceptors in the cardiovascular effects of moxonidine.

The central sympathoinhibition caused by moxonidine has been explained by activation of alpha 2-adrenoceptors on the one hand, and by an action at imidazoline I1 receptors on the other hand. In order to examine these possibilities, effects of moxonidine were compared with those of 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14304), an alpha 2-adrenoceptor agonist with very low affinity for I1 receptors, in conscious rabbits. The interaction with yohimbine, an alpha 2-adrenoceptor antagonist with very low affinity for imidazoline I1 receptors, was also studied. Moxonidine 3-100 micrograms kg-1 and UK 14304 1-30 micrograms kg-1 i.v. elicited similar effects: they decreased arterial blood pressure after a transient increase, decreased renal sympathetic nerve activity (recorded with chronically implanted electrodes), decreased heart rate and decreased the plasma noradrenaline concentration. Yohimbine given i.v. antagonized the effects of moxonidine and of UK 14304 in a similar manner. Yohimbine injected into the cisterna magna (i.c.) prevented the hypotension but did not change the decrease in plasma noradrenaline and heart rate, again in the case of both moxonidine and UK 14304. The agreement of the effect patterns of moxonidine and UK 14304, and the similar antagonism of yohimbine against either drug, demonstrate involvement of alpha 2-adrenoceptors in their central sympathoinhibitory action. The resistance of the bradycardia and the plasma noradrenaline fall against yohimbine i.c. indicates a contribution of presynaptic alpha 2-adrenergic inhibition of transmitter release from postganglionic sympathetic neurons to the overall reduction of sympathetic tone.

Adrenergic alpha-2 Receptor Agonists↗
PubMed1995

Kainate receptors are involved in the glutamate-induced indirect, purinergic inhibition of [3H]-noradrenaline release in rabbit brain cortex.

Activation of ionotropic but not metabotropic glutamate receptors causes an indirect inhibition of the release of noradrenaline in slices of rabbit brain cortex. The inhibition is mediated by adenosine which activates presynaptic adenosine A1-receptors. The present study characterizes the ionotropic receptor types through which glutamate itself produces this indirect inhibition. Rabbit brain cortex slices were preincubated with [3H]-noradrenaline, superfused with medium containing desipramine (1 microM) and stimulated electrically by trains of 6 pulses at 100 Hz. Glutamate (100-3000 micro M) reduced the electrically evoked overflow of tritium by up to 58%. The effect did not differ 20 min and 60 min after addition of glutamate. Adenosine deaminase (1 U ml-1) as well as 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 30 microM) and D-gamma-glutamylamino-methanesulfonate (GAMS; 30 micro M), both of which block kainate receptors, attenuated the glutamate-induced inhibition. The NMDA receptor antagonist 2-amino-5-phosphonopentanoate (AP5; 100 micro M) and the AMPA receptor antagonist 6-nitro-7-sulfamoylbenzo(f)-quinoxaline-2,3-dione (NBQX; 30 micro M) did not change the effect of glutamate. Given alone, CNQX and GAMS, but not AP5 and NBQX, slightly increased the evoked overflow of tritium; the increases were abolished in the presence of adenosine deaminase. The results indicate that activation of kainate but not NMDA and AMPA receptors is involved in the indirect, adenosine-mediated inhibition by exogenous glutamate of the release of noradrenaline in rabbit brain cortex slices. Moreover, as shown by the increase caused by CNQX and GAMS, endogenous excitatory amino acids inhibit the release of noradrenaline through the kainate receptor-adenosine mechanism and thus contribute to the purinergic inhibitory control of noradrenaline release in the brain.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

Two relaxation-mediating P2-purinoceptors in guinea-pig taenia caeci.

2-Methylthio ATP, noradrenaline, adenosine 5'-O-(2-thiodiphosphate) (ADP beta S), alpha, beta-methylene ATP, ATP and adenosine caused relaxation of guinea-pig isolated taenia caeci precontracted by carbachol, with potency decreasing in the order indicated. 4,4'-Diisothiocyanatostilbene-2,2'-disulphonate (DIDS) 10, 32 and 100 microM shifted the concentration-response curve of alpha, beta-methylene ATP increasingly to the right, by the factor 141 at DIDS 100 microM. Concentration-response curves of the other agonists were not shifted to the right by DIDS 100 microM, except for the curve of ADP beta S which was shifted by the factor 2.4. The relaxation produced by 2-methylthio ATP faded rapidly. When the fade was complete, further addition of 2-methylthio ATP or ATP did not elicit relaxation, whereas the relaxant effect of alpha, beta-methylene ATP was unchanged. The results indicate that there are at least two relaxation-mediating P2-purinoceptors in guinea-pig taenia caeci, the P2Y-purinoceptor which is relatively insensitive to DIDS and a distinct receptor for alpha, beta-methylene ATP which is very sensitive to DIDS.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1995

Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2A-autoreceptors in rabbit atria and kidney.

The study was devised to classify, by means of antagonist affinities, the presynaptic alpha 2-autoreceptors of rabbit atria and kidney in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists was chosen that was able to discriminate between the four subtypes. Small pieces of the left atrium and slices of the kidney cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. In one series of experiments, tissue pieces were stimulated by relatively long pulse trains (1 or 2 min) leading to alpha 2-autoinhibition. All 11 (atria) or 10 (kidney) antagonists increased the evoked overflow of tritium. pEC30% values (concentrations causing 30% increase) were interpolated from concentration-response curves. In a second series of experiments, tissue pieces were stimulated by brief pulse trains (0.4 s) that did not lead to alpha 2-autoinhibition, and concentration-inhibition curves of the alpha 2-selective agonist 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) were determined. Most of the 11 (atria) or 8 (kidney) antagonists shifted the concentration-inhibition curve of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. pEC30% values correlated with pKd values, both in atria (r = 0.728) and in the kidney (r = 0.930). pEC30% values in atria correlated with pEC30% values in the kidney (r = 0.988) and pKd values in atria correlated with pKd values in kidney (r = 0.923). It is concluded that the alpha 2-autoreceptors in atria and the kidney are the same. Comparison with antagonist affinities for prototypic native alpha 2 binding sites, alpha 2 binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature--indicates that both autoreceptors are alpha 2A. This conclusion is reached with either of the two independent estimates of autoreceptor affinity, pEC30% and pKd. The results are compatible with the hypothesis that at least the majority of alpha 2-autoreceptors belong to the alpha 2A/D branch of the alpha 2-adrenoceptor tree, across mammalian or at least rodent and lagomorph species.

Animals↗
PubMed1995

Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors in mouse brain.

The study was devised to classify, by means of antagonist affinities, the presynaptic alpha 2-autoreceptors in mouse cerebral cortex in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists was chosen that was able to discriminate between the four subtypes. Slices of the cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. The stimulation periods used (4 pulses, 100 Hz) did not lead to alpha 2-autoinhibition as shown by the lack of an increase by rauwolscine of the evoked overflow of tritium. The alpha 2-selective agonists 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) and alpha-methylnoradrenaline reduced the evoked overflow. All 10 antagonists shifted the concentration-inhibition curve of UK 14,304 to the right. Rauwolscine also shifted the concentration-inhibition curve of alpha-methylnoradrenaline to the right. pKd values of the antagonists were calculated from the shifts. The pKd values of rauwolscine against UK 14,304 and alpha-methylnoradrenaline were very similar (8.0 and 7.9, respectively). Comparison with antagonist affinities for prototypic native alpha 2 binding sites, alpha 2 binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature--indicates that the alpha 2-autoreceptors in mouse brain cortex are alpha 2D. This is the first subtype determination of alpha 2-autoreceptors in the mouse. It supports the hypothesis that at least the majority of alpha 2-autoreceptors belong to the alpha 2A/D branch of the alpha 2-adrenoceptor tree.

Adrenergic alpha-Agonists↗
PubMed1995

Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors in guinea-pig atria and brain.

The study was devised to classify, by means of antagonist and agonist affinities, the presynaptic alpha 2-autoreceptors in guinea-pig heart atria and brain cortex in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D. A set of antagonists and agonists was chosen that was able to discriminate between the four subtypes. Small pieces of the atria and slices of the brain cortex were preincubated with 3H-noradrenaline and then superfused and stimulated electrically. In one series of experiments (atria only), tissue pieces were stimulated by relatively long pulse trains (1 min) leading to marked alpha 2-autoinhibition. All 10 antagonists increased the evoked overflow of tritium. pEC30% values (concentrations causing 30% increase) were interpolated from concentration-response curves. In a second series of experiments (atria and brain slices), tissue pieces were stimulated by brief pulse trains (0.4 s or 40 ms) that led to little (atria) or no (brain slices) alpha 2-autoinhibition, and antagonist effects against the alpha 2-selective agonist 5-bromo-6-(2-imidazolin-2- ylamino)-quinoxaline (UK 14,304) were examined. All 10 (atria) or 8 (brain) antagonists shifted the concentration-inhibition curve of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. In a third series of experiments (brain slices only), also with brief pulse trains (40 ms), pKa values (negative logarithms of dissociation constants of agonist-alpha 2-adrenoceptor complexes) were determined by comparison of concentration-inhibition curves of UK 14,304, guanoxabenz and oxymetazoline in normal tissue and in tissue in which a fraction of the receptors had been blocked by phenoxybenzamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenergic alpha-Agonists↗
PubMed1995

Subclassification of presynaptic alpha 2-adrenoceptors: alpha 2D-autoreceptors and alpha 2D-adrenoceptors modulating release of acetylcholine in guinea-pig ileum.

The study was designed to classify in terms of alpha 2A, alpha 2B, alpha 2C and alpha 2D the presynaptic alpha 2-autoreceptors, as well as the alpha 2-receptors modulating the release of acetylcholine, in the myenteric plexus-longitudinal muscle (MPLM) preparation of the guinea-pig ileum. A set of antagonists was chosen that was able to discriminate between the four subtypes. Small pieces of the MPLM preparation were preincubated with 3H-noradrenaline or 3H-choline and then superfused and stimulated electrically. The stimulation periods used (3H-noradrenaline: 3 trains of 20 pulses, 50 Hz, train interval 60 s; 3H-choline: single trains of 30 pulses, 0.2 Hz) did not lead to alpha 2-autoinhibition or inhibition of 3H-acetylcholine release by endogenous noradrenaline. The alpha 2-selective agonist 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14,304) reduced the evoked overflow of tritium in both 3H-noradrenaline and 3H-choline experiments. Most (3H-noradrenaline) or all (3H-choline) of the 10 antagonists shifted the concentration-inhibition curves of UK 14,304 to the right. pKd values of the antagonists were calculated from the shifts. pKd values from 3H-noradrenaline experiments correlated with pKd values from 3H-choline experiments (r = 0.981). It is concluded that alpha 2-autoreceptors and alpha 2-heteroreceptors modulating the release of acetylcholine in the MPLM preparation are of the same subtype. Comparison with antagonist affinities for prototypic native alpha 2 binding sites, binding sites in cells transfected with alpha 2 subtype genes, and previously classified presynaptic alpha 2-adrenoceptors--all taken from the literature--indicates that both are alpha 2D.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1995

Reactive red 2: a P2y-selective purinoceptor antagonist and an inhibitor of ecto-nucleotidase.

Effects of reactive red 2 and its parent compound acid red 33 were studied in rat vas deferens and guinea-pig taenia coli. In rat vas deferens, reactive red 2(1 to 10 microM) shifted the concentration-response curve for the P2X-purinoceptor-mediated contraction effect of alpha, beta-methylene ATP slightly to the right and progressively decreased the maximum (apparent antagonist Kd value 0.42 microM). Acid red 33 (1000 microM) shifted the curve to the right without changing the maximum (apparent Kd 386 microM). The concentration-contraction curve of noradrenaline was not altered by reactive red 2. In the carbachol-precontracted guinea-pig taenia coli, reactive red 2 (0.1 to 10 microM) shifted the concentration-response curve for the P2Y-purinoceptor-mediated relaxation effect of adenosine 5'-O-(2-thiodiphosphate) (ADP beta S) progressively to the right; only at the highest concentration of antagonist (10 microM) was the maximum slightly depressed; a pA2 value of 7.55 (Kd 0.028 microM) was derived from the shift. Acid red 33 (1000 microM) shifted the concentration-relaxation curve of ADP beta S to the right without changing the maximum (apparent Kd 171 microM). Reactive red 2 (1 to 10 microM) also shifted the concentration-response curve for the relaxation effect of alpha, beta-methylene ATP, which is mediated by an unclassified P2-purinoceptor, progressively to the right but simultaneously decreased the maximum (apparent Kd 1.6 microM). The concentration-relaxation curve of 2-chloroadenosine was not altered by reactive red 2. Pieces of vas deferens and taenia coli degraded 76 and 66% of added ATP (10 microM) within 30 min, respectively. Reactive red 2(0.1 to 100 microM) progressively reduced this degradation by up to 95%, with IC50 values of 3.9 +/- 0.6 and 3.9 +/- 2.3 microM, respectively. Acid red 33 (1000 microM) reduced the degradation by 30 and 20%, respectively. The results indicate that reactive red 2 is a relatively potent antagonist at both P2X-purinoceptors in rat vas deferens and P2Y-purinoceptors in guinea-pig taenia coli, with a 15 fold selectively for the P2Y-purinoceptor. It inhibits ecto-nucleotidase in both tissues. The dichloro-triazine residue that distinguishes the compound from acid red 33 greatly enhances the potency at both receptor subtypes as well as at the nucleotidase. As regards P2-purinoceptor subtypes, the results confirm the existence of two relaxation-mediating P2-purinoceptors in guinea-pig taenia coli.

Adenosine Triphosphatases↗
PubMed1995

Functional consequences of inhibition of nucleotide breakdown in rat vas deferens: a study with Evans blue.

The effect of Evans blue on nucleotide breakdown, nucleotide-evoked contractions and electrically evoked contractions, overflow of ATP and overflow of tritium (after labelling with [3H]-noradrenaline) was studied in rat vas deferens. Pieces of vas deferens degraded 83 to 85% of added ATP, ADP and 2-methyl-thio ATP (all 100 microM) over 30 min. Evans blue (100 microM) reduced this degradation to 22 to 26%. Nucleotides elicited contraction with potency declining in the order alpha,beta-methylene ATP > 2-methylthio ATP > ATP > ADP. Evans blue (100 microM) shifted the concentration-response curve of alpha, beta-methylene ATP to the right and increased the maximum. Concentration-response curves of ATP, ADP and 2-methylthio ATP, in contrast, were shifted to the left and responses were much potentiated. In the presence of Evans blue, the rank order of potency was ATP > 2-methylthio ATP > alpha, beta-methylene ATP > ADP. Electrical field stimulation (100 pulses at 10 Hz) elicited contraction and an overflow of tritium and ATP. Evans blue (100 microM) did not alter the contraction and the evoked overflow of tritium but increased 24-fold the evoked overflow of ATP. The results indicate that Evans blue may serve as an-albeit impure-ecto-nucleotidase inhibitor in functional experiments. Such experiments demonstrate that the low potency of ATP (and also ADP and 2-methylthio ATP) in eliciting contraction, and the small size of the overflow of ATP upon sympathetic nerve stimulation, are due to rapid breakdown.

Adenosine Triphosphate↗
PubMed1995

Comparison of corelease of noradrenaline and ATP evoked by hypogastric nerve stimulation and field stimulation in guinea-pig vas deferens.

Contractions and overflow of tritium and ATP elicited by hypogastric nerve stimulation (HNS) and field stimulation (FS) were studied in the guinea-pig isolated vas deferens preincubated with [3H]-noradrenaline. ATP was measured by means of the luciferin-luciferase technique. HNS and FS elicited contraction, tritium overflow and ATP overflow. HNS at supramaximal current strength produced smaller responses than did FS at supramaximal current strength (210 pulses/7 Hz). Supramaximal HNS and submaximal FS were used in the remainder of the study. Prazosin (0.3 mumol/l) reduced contractions and the overflow of ATP elicited by both HNS and FS; the evoked overflow of tritium was not changed (210 pulses/7 Hz). Combined administration of prazosin (0.3 mumol/l) and suramin (300 mumol/l) abolished contractions and reduced the overflow of ATP elicited by both HNS and FS slightly more than did prazosin alone; tritium overflow again was not changed (210 pulses/7 Hz). Contractions, tritium overflow and ATP overflow increased with the frequency of both HNS and FS (from 7 to 25 Hz; 210 pulses); the increase in ATP overflow with frequency was more marked than the increase in tritium overflow. The preferential increase of ATP overflow with the frequency of HNS and FS persisted in the combined presence of prazosin (0.3 mumol/l) and suramin (300 mumol/l). The study confirms for HNS, a more physiologic way of sympathetic nerve stimulation, several observations previously obtained with FS. First, HNS-evoked ATP release is detectable as an overflow of ATP into the superfusion fluid.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1995

Cardiovascular effects of agmatine, a "clonidine-displacing substance", in conscious rabbits.

Agmatine has been identified as a "clonidine-displacing substance" in extracts from bovine brain. We studied its effect on cardiovascular regulation and the role played in this effect by alpha 2-adrenoceptors. In conscious rabbits, agmatine 10 micrograms kg-1 injected intracisternally (i.c.) caused no change, whereas agmatine 30, 100 and 300 micrograms kg-1 i.c. increased renal sympathetic nerve firing, the plasma concentration of noradrenaline and adrenaline and arterial blood pressure. Heart rate tended to be decreased. Yohimbine 1.5 micrograms kg-1 i.c. caused no change, whereas yohimbine 5, 15 and 50 micrograms kg-1 increased renal sympathetic nerve activity, the plasma concentration of noradrenaline and adrenaline, blood pressure and heart rate. In rabbit brain cortex slices preincubated with [3H]-noradrenaline, agmatine 1 to 100 microM did not modify the electrically evoked overflow of tritium (either 4 pulses at 100 Hz or 36 pulses at 3 Hz). The evoked overflow was reduced by 5-bromo-6-(2-imidazolin-2-ylamino)-quinoxaline (UK 14304) 0.03 to 30 nM (4 pulses at 100 Hz), and this inhibition was not affected by agmatine 10 and 100 microM. Agmatine did not change the basal efflux of tritium. The results show that agmatine, like yohimbine, causes central sympathoexcitation when given i.c., but agmatine differs from yohimbine in that it does not increase heart rate. Agmatine acts neither as an agonist nor as an antagonist at the alpha 2-autoreceptors in rabbit brain cortex. alpha 2-Adrenoceptors, therefore, are probably not involved in its cardiovascular effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1995

P2-purinoceptor-mediated inhibition of noradrenaline release in rat atria.

1. We looked for P2-purinoceptors modulating noradrenaline release in rat heart atria. Segments of the atria were preincubated with [3H]-noradrenaline and then superfused with medium containing desipramine (1 microM) and yohimbine (1 microM) and stimulated electrically, by 30 pulses/1 Hz unless stated otherwise. 2. The adenosine A1-receptor agonist, N6-cyclopentyl-adenosine (CPA; EC50 9.7 nM) and the nucleotides, ATP (EC50 6.6 microM) and adenosine-5'-O-(3-thiotriphosphate) (ATP gamma S; EC50 4.8 microM), decreased the evoked overflow of tritium. The adenosine A2a-agonist, 2-p-(2-carbonylethyl)-phenethylamino-5'-N-ethylcarboxamido-a denosine (CGS-21680; 0.03-0.3 microM) and the P2x-purinoceptor agonist beta, gamma-methylene-L-ATP (30 microM) caused no change. 3. The concentration-response curve of CPA was shifted to the right by the adenosine A1-receptor antagonist, 8-cyclopentyl-1,3-dipropyl-xanthine (DPCPX; 3 nM; apparent pKB value 9.7) but hardly affected by the P2-purinoceptor antagonist, cibacron blue 3GA (30 microM). In contrast, the concentration-response curves of ATP and ATP gamma S were shifted to the right by DPCPX (3 nM; apparent pKB values 9.3 and 9.4, respectively) as well as by cibacron blue 3GA (30 microM; apparent pKB values 5.0 and 5.1, respectively). Combined administration of DPCPX and cibacron blue 3GA caused a much greater shift of the concentration-response curve of ATP than either antagonist alone. The concentration-response curve of ATP was not changed by indomethacin, atropine or the 5'-nucleotidase blocker alpha, beta-methylene-ADP. 4. Cibacron blue 3GA (30 microM) increased the evoked overflow of tritium by about 70%. The increase was smaller when the slices were stimulated by 9 pulses/O00 Hz instead of 30 pulses/I Hz.5. The results indicate that the postganglionic sympathetic axons in rat atria possess P2-purinoceptors in addition to the known adenosine Al-receptor. Both mediate inhibition of noradrenaline release. Some adenine nucleotides such as ATP and ATP gamma S act at both receptors. The presynaptic P2-purinoceptor seems to be activated by an endogenous ligand, presumably ATP, under the condition of these experiments. This is the first evidence for presynaptic P2-purinoceptors at cardiac postganglionic sympathetic axons.

Adenosine↗