Actions of ATP and alpha, beta-methylene ATP on neuromuscular transmission and smooth muscle membrane of the rabbit and guinea-pig mesenteric arteries.

In the rabbit mesenteric artery, adenosine triphosphate (ATP), showed two actions on the membrane potential of muscle cells: low concentrations (1-10 microM) hyperpolarized and high concentrations (greater than or equal to 50 microM) depolarized the membrane. Both changes in the potential were accompanied by increases in ionic conductance. In the rabbit mesenteric artery, alpha, beta-methylene ATP (MeATP), (greater than or equal to 30 nM) depolarized the muscle membrane at a lower concentration than ATP (greater than or equal to 50 microM), and increased the ionic conductance of the membrane. The depolarization induced by ATP was prevented by low concentrations of MeATP, but the hyperpolarization was retained. Furthermore, the hyperpolarization was not affected by theophylline (10 microM). In the guinea-pig mesenteric artery, ATP and MeATP depolarized and increased the ionic conductance of muscle membrane, but to depolarize the membrane, higher concentrations of both agents were required, compared to those in the rabbit mesenteric artery. In the mesenteric arteries from both species, perivascular nerve stimulation evoked excitatory junction potentials (e.j.ps). In both tissues, MeATP inhibited the amplitude of e.j.ps at lower concentrations than did ATP, and both agents had more potent inhibitory actions on rabbit than on guinea-pig. The inhibition of e.j.p. induced by low concentrations of these agents showed no relationship to depolarization, but the inhibition induced by high concentrations was paralleled by depolarization and increase in ionic conductance of the membrane. In the rabbit mesenteric artery, overflow of noradrenaline (NA) and its metabolite (3,4-dihydroxyphenylglycol; DOPEG) produced by perivascular nerve stimulation was examined. ATP (0.1 mM) but not MeATP (0.1 microM) reduced the overflow of NA, whereas both agents had no effect on the overflow of DOPEG. Exogenously applied high concentrations of NA (greater than or equal to 3 microM) depolarized the muscle membrane in both species. These NA-induced depolarizations were not affected by treatment with ATP or MeATP. It is concluded that, in the rabbit mesenteric artery, ATP is more likely to be involved in generation of e.j.ps than is NA. A similar interpretation in the guinea-pig mesenteric artery is complicated by the depolarization produced by high concentrations of ATP or MeATP.     

Effects of dopamine on neuromuscular transmission in the guinea-pig mesenteric artery

The effects of dopamine on neuromuscular transmission in the guinea-pig mesenteric artery were investigated using a microelectrode method. Dopamine did not modify the membrane potential or the membrane resistance of the smooth muscle, but did reduce the amplitude of excitatory junction potentials (e.j.p.) and enhance the facilitation of e.j.p. produced by repetitive stimulation. Phentolamine (10(-7) M) enhanced the amplitude and the facilitation of the e.j.p., and with the addition of dopamine (10(-6) M) there was a reduction in the amplitude of e.j.p. but not in the facilitation. Haloperidol and sulpiride (greater than 10(-6) M) increased the amplitude of e.j.p. without altering the postjunctional membrane properties. Haloperidol and sulpiride did not increase the facilitation of e.j.p. produced by repetitive stimulation. In the presence of haloperidol or sulpiride (10(-5) M), dopamine (10(-6) M) did not suppress the amplitude of the e.j.p. These results indicate that in the guinea-pig mesenteric artery, dopamine inhibits the release of transmitter at the presynaptic membrane.     

Effects of nifedipine derivatives on smooth muscle cells and neuromuscular transmission in the rabbit mesenteric artery

The effects of nifedipine and its derivatives, nisoldipine, nimodipine and nitrendipine, on smooth muscle cells and neuromuscular transmission were investigated in the rabbit mesenteric artery. These agents in concentrations of up to 2 X 10(-7) M modified neither the membrane potential nor the membrane resistance, yet did inhibit the spike potential evoked by direct muscle stimulation in the presence of TTX or by perivascular nerve stimulation. The inhibitory action of nitrendipine was weaker than that of the other derivatives. These agents had no effect on the miniature excitatory junction potentials (m.e.j.ps) and e.j.ps evoked by the first stimuli and after completion of the facilitation in a train stimulation. Nifedipine and its derivatives had no effect on the K-induced depolarization but did have a marked effect on the K-induced contraction. Nisoldipine showed the highest inhibitory potency for the K-induced contraction [IC50 was 1.2 X 10(-9) M for the 128 mM (K)o-induced contraction]. Noradrenaline depolarized the membrane (greater than 5 X 10(-7 M) and produced contraction (greater than 3 X 10(-7) M). The contraction evoked by high concentrations of noradrenaline was inhibited by these agents to a greater extent than that evoked by low concentrations. The contraction evoked by perivascular nerve or direct muscle stimulation was partly inhibited by nifedipine and its derivatives. The contraction elicited by Na-free solution was inhibited by these agents but the noradrenaline- or caffeine-induced contraction in Ca-free solution was not. These results indicate that in smooth muscle cells of the rabbit mesenteric artery, nifedipine and its derivatives inhibit the voltage dependent Ca-influx which occurs during the spike potential and in response to K-, electrically- or noradrenaline-induced depolarization. These derivatives appear to have no effect on the adrenoceptor operated Ca increase in myoplasm which occurs in the absence of depolarization at low noradrenaline concentrations. The derivatives act as Ca antagonists with a quantitative difference in potency, i.e. the strongest action was observed with nisoldipine and the weakest with nitrendipine.     

Effects of dopamine on adrenergic neuromuscular transmission in the guinea-pig vas deferens.

1 In the isolated vas deferens of the guinea-pig, contractile responses to adrenergic nerve stimulation at 2 Hz were depressed by exogenous dopamine (5 microM) and this effect was abolished in the presence of phentolamine (0.3 microM), suggesting that it was due to an agonist action of dopamine on alpha-adrenotors. 2 The depression by dopamine (5 microM) of contractile responses to nerve stimulation was correlated with reduction in amplitude of single excitatory junction potentials (e.j.ps) evoked by nerve stimulation, but not with depression of spontaneous junction potentials. 3 By contrast, during repetitive nerve stimulation at 1 Hz the depressant effect of dopamine on e.j.p. amplitude became less pronounced, due to the amount of facilitation being greater than that occurring under control conditions in the same cell. 4 The alpha-adrenoceptor antagonist, phentolamine (10 microM), also increased the amount of facilitation during repetitive nerve stimulation. 5 In the presence of phentolamine (10 microM), the depressant effect of dopamine (5 microM) on single e.j.ps was abolished but its enhancing effect on facilitation was not reduced. 6 It is suggested that the enhancement of facilitation during repetitive stimulation by both dopamine and phentolamine is independent of their actions on presynaptic alpha-adrenoceptors.     

Pinacidil inhibits the ryanodine-sensitive outward current and glibenclamide antagonizes its action in cells from the rabbit portal vein.

Pinacidil, a potassium-channel opener, inhibited the ryanodine-sensitive oscillatory outward potassium current induced by Ca released from an intracellular store. Glibenclamide, a blocker of the ATP-sensitive K-channel, prevented the action of pinacidil, suggesting the presence of an additional site (to K channels) for the vasodilator actions of pinacidil at which glibenclamide can act as an antagonist.     

Effects of prostaglandin I2 and carbocyclic thromboxane A2 on smooth muscle cells and neuromuscular transmission in the guinea-pig mesenteric artery

1 In the guinea-pig mesenteric arteries neither prostacyclin (PGI₂) nor carbocyclic thromboxane A₂ (cTxA₂) affected membrane potential in concentrations below 1 × 10^-6 M. Increasing the concentration to 3 × 10^-6 M either slightly hyperpolarized or depolarized the membrane with little change in membrane resistance.

2 At a concentration of 1 × 10^-7 M, the amplitude of the first e.j.p. and the enlarged amplitudes of the subsequent e.j.ps evoked by trains of stimuli were reduced consistently by PGI₂ or cTxA₂. Facilitation was unaffected by either agent.

3 The inhibitory actions of PGI₂ were partly overcome by increased concentrations of 5 mM [Ca]_o and were accelerated by a reduced concentration of 1.25 mM [Ca]_o.

4 The amplitude of the contraction evoked by perivascular nerve stimulation was inhibited to a greater extent by PGI₂ than by cTxA₂ at concentrations below 1 × 10^-6 M.

5 The contraction evoked by 5 × 10^-6 M noradrenaline (NA) or excess concentrations of 20.2 mM [K]_o was enhanced by 1 × 10^-8 M — 1 × 10^-6 M cTxA₂ and suppressed by 1 × 10^-8 M — 1 × 10^-6 M PGI₂. The minimum concentration of cTxA₂ required to produce the contraction was 1 × 10^-8 M.

6 These results indicate that transmission at the neuromuscular junction was inhibited consistently by PGI₂ or cTxA₂, presumably due to inhibition of NA release by suppression of the Ca influx at the nerve terminals. Whereas PGI₂ inhibited, cTxA₂ enhanced the mechanical response by a direct action on the smooth muscle cells.

     

The distribution of gamma-adrenoceptors and P2 purinoceptors in mesenteric arteries and veins of the guinea-pig.

1. Membrane potential changes and contractions were recorded from mesenteric arteries and veins of the guinea-pig, during perivascular nerve stimulation or application of noradrenaline or adenosine triphosphate (ATP). 2. After alpha-adrenoceptor blockade, noradrenaline activated low affinity adrenoceptors (gamma-adrenoceptors) causing depolarization and arterial contraction only in the presence of an inhibitor of catecholamine uptake. 3. Noradrenaline did not cause depolarization or contraction of the vein after alpha-adrenoceptor blockade even after catecholamine uptake was blocked. 4. Adenosine triphosphate caused depolarization and contraction of both arteries and veins. These responses were abolished by alpha-,beta-,methylene adenosine triphosphate (Me-ATP). 5. Me-ATP abolished rapid excitatory junction potentials (e.j.ps) caused by perivascular nerve stimulation of arteries but had no effect on arterial responses mediated by gamma-adrenoceptors. 6. In veins, perivascular nerve stimulation evoked slow e.j.ps which persisted in the presence of Me-ATP but were abolished after blockade of alpha-adrenoceptors. 7. The observations indicate that P2 purinoceptors are present on both mesenteric artery and vein whilst gamma-adrenoceptors are localized near the neuromuscular junction of the artery. However gamma-adrenoceptors do not appear to be directly involved in the generation of arterial e.j.ps.     

Co-release of endogenous ATP and noradrenaline from guinea-pig mesenteric veins exceeds co-release from mesenteric arteries

1. The present study was designed to compare the overflow of sympathetic neurotransmitters of guinea-pig inferior mesenteric artery and mesenteric vein evoked by electrical field stimulation (EFS) with special emphasis on the simultaneous release of ATP and noradrenaline (NA). The stimulation-evoked overflow of ADP, AMP and adenosine was also evaluated. 2. Endothelium-denuded segments of inferior mesenteric arteries or veins were superfused in a small volume (200 microL)-chamber for EFS and subsequent detection of NA (HPLC- electrochemical detection) and adenine nucleotides and adenosine (HPLC-fluorescence detection) in samples of the superfusate. 3. Both arteries and veins responded to EFS (15 V, 4-16 Hz, 0.3 msec for 60 s) with overflow of ATP and NA in a tetrodotoxin (1 micromol/L)- and guanethidine (10 micromol/L)-sensitive manner. The EFS-evoked overflow of NA in veins exceeded the overflow of NA in arteries at all frequencies of stimulation, whereas the EFS-evoked overflow of ATP, ADP and AMP in veins exceeded the overflow of adenine nucleotides in arteries at 4 and 8 Hz but not at 16 Hz stimulation. The EFS-evoked overflow of adenosine was similar in arteries and veins. 4. Activation of alpha1-adrenoceptors with methoxamine (10 micromol/L) did not produce overflow of ATP. 5. Blockade of alpha1/alpha2-adrenoceptors with phentolamine (1 micromol/L) did not affect EFS-evoked overflow of ATP, ADP, AMP and adenosine. 6. It is concluded that overflow of ATP and NA from sympathetic nerves may constitute an effective mechanism in the complex balance between capacitance and resistance in splanchnic circulation.     

The role of endothelium in the phenylephrine-induced oscillatory responses of rabbit mesenteric arteries.

Phenylephrine-induced oscillatory contractions in rabbit mesenteric arteries were investigated in vitro. Adrenergic, cholinergic, or histamine antagonists as well as cyclooxygenase and lipoxygenase inhibitors had no effect on this phenylephrine-induced oscillation. The removal of extracellular calcium ions or treatment with a calcium antagonist reduced the amplitude and frequency of the oscillation. Removal of the endothelium or treatment with inhibitors of the synthesis or the target enzyme of endothelium-derived relaxing factor (EDRF) also reduced the amplitude and frequency of the oscillation. In a perfusion bioassay, the perfusate from an endothelium-intact arterial segment induced oscillation of an endothelium-denuded arterial ring recipient. These results suggest that phenylephrine-induced oscillation is mediated by an endothelium-derived factor such as EDRF and depends on the influx of extracellular calcium ions.     

Modulation by noradrenaline and yohimbine of noradrenergic transmission in the guinea-pig mesenteric artery

In the guinea-pig mesenteric artery, transmitter release modulated by noradrenaline (NA) or yohimbine was estimated from changes in amplitude of the excitatory junction potential (e.j.p.) recorded from smooth muscle cells. NA decreased the amplitude of the e.j.p. with no change in the facilitation. Yohimbine antagonized the effect of NA on the e.j.p. amplitude and enhanced the facilitation of e.j.p.; the latter action was not antagonized by NA. TTX-resistant e.j.p.s evoked by stronger intensity of stimuli were not affected by NA or yohimbine. It is concluded that NA inhibits and yohimbine enhances the release of transmitter, and that the latter event involves prejunctional alpha-adrenoceptor-dependent and -independent processes.     

5-Hydroxytryptamine receptors that facilitate excitatory neuromuscular transmission in the guinea-pig isolated detrusor muscle.

1. In isolated detrusor strips from the guinea-pig urinary bladder, contractile responses to electrical field stimulation were mostly mediated by neurally released acetylcholine (ACh) and adenosine 5'-triphosphate (ATP). 2. 5-Hydroxytryptamine (5-HT) produced a concentration-dependent increase in the amplitude of stimulated detrusor strip contractions. The 5-HT concentration-response curve showed a biphasic profile: the high potency phase was obtained at sub-micromolar concentrations (10-300 nM), while the low potency phase in the range 1-30 microM. The maximum response of the first phase was 30% of the total 5-HT response. 3. Like 5-HT, the 5-HT3 receptor agonist, 2-methyl-5-hydroxytryptamine (2-methyl-5-HT: 0.3-100 microM), the 5-HT2 receptor agonist, (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI: 30 nM-3 microM) and the 5-HT4 receptor agonist, 5-methoxytryptamine (5-MeOT: 0.1-30 microM) potentiated, though with lower potency, detrusor contractions. The resulting concentration-response curves were monophasic in nature. 2-Methyl-5-HT had a maximum effect comparable to that of 5-HT. By contrast, the maximal effects of DOI and 5-MeOT were only 20% and 30% of that elicited by 30 microM 5-HT, respectively. 4. The 5-HT3 receptor antagonist, granisetron (0.3 microM) had no effect on the high potency phase, but caused a rightward parallel shift of the low potency phase of the 5-HT curve (pKB = 7.3).(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of the effects of caffeine and procaine on noradrenergic transmission in the guinea-pig mesenteric artery.

The effects of caffeine and procaine on noradrenergic transmission in the guinea-pig mesenteric artery were investigated by recording electrical responses of smooth muscle cells and by measuring the outflow of noradrenaline (NA) and 3,4-dihydroxyphenylglycol (DOPEG) induced by perivascular nerve stimulation. Caffeine possessed dual actions on the membrane, i.e., at low concentrations (2.5 X 10(-4)-5 X 10(-4)M), it hyperpolarized the membrane and decreased the membrane resistance and at high concentrations (over 2.5 X 10(-3)M) it depolarized the membrane and increased the membrane resistance. Procaine (over 10(-4)M) consistently depolarized the membrane and increased the membrane resistance. The amplitude of the excitatory junction potential (e.j.p.) produced by perivascular nerve stimulation was increased by low concentrations of procaine (2.5 X 10(-5)-10(-4)M) or high concentrations (10(-3)-5 X 10(-3)M) of caffeine and was decreased by low concentrations of caffeine (2.5 X 10(-5)-10(-4)M) or high concentrations of procaine (5 X 10(-4)-10(-3)M). Higher concentrations of caffeine (over 5 X 10(-3)M) induced a spike potential on the e.j.p., while higher concentrations of procaine (over 2.5 X 10(-3)M) inhibited the generation of e.j.ps. Facilitation of e.j.ps produced by repetitive stimulation of perivascular nerves remained unchanged by caffeine, while it was enhanced by procaine at any given concentration (caffeine 2.5 X 10(-4)-10(-3)M; procaine 10(-4)-10(-3)M). The membrane depolarization produced by exogenously applied NA (10(-5)M) was not blocked by pretreatment with procaine. Conduction velocity of perivascular nerve excitation remained unchanged by application of caffeine (up to 5 X 10(-3)M), and was reduced by application of procaine (over 2.5 X 10(-4)M). Outflow of NA during perivascular nerve stimulation remained unchanged by caffeine (10(-4)-3 X 10(-3)M), while it was enhanced by procaine (over 2.5 X 10(-4)M). The outflow of DOPEG was slightly reduced by caffeine (10(-3)-5 X 10(-3)M) and by lower concentrations of procaine (10(-4)-2.5 X 10(-4)M) but was not altered by higher concentrations of procaine (10(-3)-5 X 10(-3)M). It is concluded that in the guinea-pig mesenteric artery, high concentrations of caffeine (over 10(-3)M) increased the e.j.p. amplitude which might be due to an increase in membrane resistance of the smooth muscle cells. No marked effect of caffeine was observed on transmitter release from the nerve terminals. Procaine (over 2.5 X 10(-4)M) increased transmitter release from perivascular nerves and blocked the re-uptake mechanism of released NA.(ABSTRACT TRUNCATED AT 400 WORDS)     

Pinacidil uptake and effects in the isolated rabbit heart

The myocardial accumulation of pinacidil showed one-compartment characteristics with a half-time of 1.11 min., whereas the disposition followed three-compartment kinetics with half-times for the relevant two redistributory and the terminal phases of 0.39, 1.51 and 5.44 min., respectively. At a steady-state drug concentration in the perfusate of 6.12 nmol ml-1, the average concentration of pinacidil in the myocardium was 20.6 nmol g-1. The accumulated amount could predictically be referred with 57% to a central and 31 and 12% to two peripheral (deeper) drug pools. The pharmacodynamic effects of pinacidil in the isolated perfused rabbit heart were studied at stepwise increasing concentrations from 0.15 to 100 microM. Coronary flowrate increased initially up to 24.5% at 1.5 microM pinacidil and then gradually decreased. Amplitude and velocity of contraction were both inhibited in a biphasic way up to 92.7 and 94.1%, respectively. Apparent dynamic steady states developed within 13-15 min. The computer-derived inhibitory Em-values related to the first phase were 49.2 and 52.4% and those related to the second phase were 111.7 and 108.3%, respectively. Heart frequency decreased monophasically and exhibited an inhibitory Em-value of 19.6%. Oxygen consumption decreased at pinacidil concentrations higher than 15 microM and the Em-value was 69.7%. The frequency-corrected QT-interval decreased biphasically and the related inhibitory Em-values were 8.6 and 58.7%. The QRS-interval did not change and the PQ-interval only showed a minor increase at the highest pinacidil concentration. Our findings are compatible with the concept of pinacidil being a potassium channel opener.     

Relaxation and decrease in [Ca2+]i by hydrochlorothiazide in guinea-pig isolated mesenteric arteries.

1. We examined the effect of the thiazide diuretic, hydrochlorothiazide, on on intracellular calcium concentration ([Ca2+]i) and tone in guinea-pig mesentery arteries. Vessels were mounted on a microvascular myograph and loaded with the Ca(2+)-sensitive fluorescent dye, Fura-2. 2. Hydrochlorothiazide caused relaxation of noradrenaline-precontracted arteries associated with a fall in [Ca2+]i. Preincubation of arteries with hydrochlorothiazide inhibited both contraction and rise in [Ca2+]i in response to noradrenaline. Hydrochlorothiazide did not affect tone and [Ca2+]i when this was elevated by a combination of depolarizing potassium solution and noradrenaline. 3. Hydrochlorothiazide-induced vasorelaxation and decrease of [Ca2+]i was abolished by charybdotoxin, a blocker of large conductance Ca(2+)-activated K channels. 4. The rise in [Ca2+]i elicited by caffeine in Ca(2+)-free physiological salt solution, and presumably reflecting Ca2+ release from intracellular stores, was not altered by preincubation with hydrochlorothiazide. 5. Under depolarizing conditions hydrochlorothiazide did not alter the relationship between the extracellular concentration of Ca2+ and [Ca2+]i; however, hydrochlorothiazide caused a small reduction in the contraction produced for a given rise in [Ca2+]i suggesting hydrochlorothiazide may cause a slight desensitization of the contractile machinery. 6. These findings suggest that hydrochlorothiazide opens Ca(2+)-activated K channels leading to hyperpolarization and consequent closing of voltage-operated calcium channels. The result of this is an impaired influx of extracellular Ca2+, a decrease in [Ca2+]i and vasorelaxation.     

Modification by choline of adrenergic transmission in rat mesenteric arteries

1. The action of choline on the vasoconstrictor responses of the perfused mesenteric arteries of the rat to sympathetic nerve stimulation and to injected noradrenaline has been investigated.2. The infusion of choline (500 mug/ml), for periods of 15 s, increased the response to sympathetic nerve stimulation, whereas the infusion of the same concentration for 20 min greatly reduced the response to nerve stimulation. Choline (up to 500 mug/ml), infused either for short or long periods, did not alter the response to injected noradrenaline.3. The inhibitory action of choline on the response to nerve stimulation was abolished either by an increase in the calcium concentration from 1.8 to 5.4 mM or by simultaneous infusion of (+)-amphetamine or atropine.4. The results suggest that choline in concentrations of 500 mug/ml has the same effect on adrenergic transmission in mesenteric arteries as acetylcholine at concentrations of 5 ng/ml.     

Role of EDHF in the vasodilatory effect of loop diuretics in guinea-pig mesenteric resistance arteries

1. Relaxing effect of loop diuretics, piretanide and furosemide in comparison with acetylcholine (ACh) was investigated in guinea-pig isolated mesenteric resistance arteries. 2. Concentration-response curves to ACh (0.001 - 10 microM) and diuretics (0.0001 - 1 microM) were constructed in noradrenaline (10 - 30 microM)-precontracted arteries incubated either in normal physiological salt solution (PSS) or in 30 mM KCl PSS (K-PSS). 3. In PSS, maximal relaxations (R(max)) and pD(2) to ACh were 87+/-2% and 7.1+/-0.1 (n=10). L-N(G)-nitro-arginine methyl ester (L-NAME, 100 microM) reduced R(max) by 20% (P<0.01, n=7) and pD(2) by 10% (P<0.01). In contrast, indomethacin (10 microM) increased R(max) by 19% (P<0.01, n=8) and pD(2) by 10% (P<0.05). Combination of L-NAME+indomethacin reversed the effect observed with either of these inhibitors used alone. In K-PSS, R(max) was attenuated by 40% (P<0.001, n=6) compared to PSS. L-NAME reduced R(max) by 65% (P<0.01, n=5) and increased pD(2) by 15 fold. L-NAME+indomethacin suppressed the resistant relaxation. 4. In PSS+L-NAME+indomethacin, inhibitors of small (SK(Ca); apamin, 0.1 microM) and large (BK(Ca); iberiotoxin and charybdotoxin, 0.1 microM) conductance Ca(2+)-sensitive K(-)-channels used alone had little effect on the ACh-response. Combination of apamin+iberiotoxin reduced R(max) by 40% (P<0.05, n=7) while apamin+charybdotoxin fully abolished the resistant relaxation. 5. In PSS, piretanide and furosemide induced relaxation with R(max): 89+/-3% vs 84+/-5% and pD(2): 8.5+/-0.1 vs 7.7+/-0.2 (P<0.01) for piretanide (n=11) and furosemide (n=10), respectively. Endothelial abrasion suppressed relaxation to diuretics. L-NAME and indomethacin used alone or in combination did not significantly modify the response to diuretics. 6. In K-PSS, piretanide-induced relaxation was abolished whereas that to furosemide was reduced by 70% (P<0.001, n=9) compared to PSS and was suppressed by L-NAME+indomethacin. In PSS+L-NAME+indomethacin, apamin slightly reduced relaxation to diuretics whereas charybdotoxin or iberiotoxin abolished the response. 7. These results indicate that ACh-evoked relaxation is mediated by both NO/PGl(2)-dependent and -independent mechanisms. The EDHF-dependent component relies on activation of Ca(2+)-activated K(+) channels, is sensitive to a combination of apamin+charybdotoxin and to a smaller degree to a combination of apamin+iberiotoxin. Loop diuretic-induced relaxation is endothelium-dependent, appears to be mediated by NO, PGl(2) and EDHF for furosemide and EDHF only for piretanide. For the two diuretics, opening of BK(Ca) channels may be involved in the relaxation.     

Effects of pimozide on noradrenergic transmission in rabbit isolated ear arteries.

In the rabbit ear artery both dopamine and noradrenaline inhibit stimulation-induced (S-I) transmitter noradrenaline efflux. Pimozide, which is reported to be a specific dopamine receptor antagonist, was used to further study the effects of dopamine on transmitter efflux. In a concentration of 0.2 micrometer pimozide blocked the inhibition of S-I efflux produced by 0.5 micrometer dopamine but not that produced by 0.5 micrometer noradrenaline. In a concentration of 10 nM, pimozide enhances transmitter release and vasoconstrictor responses to sympathetic nerve stimulation; this may be due to blockade of feedback inhibition of transmitter release by endogenous dopamine. In a concentration of 1 micrometer, pimozide reduced transmitter release and vasoconstrictor responses to sympathetic nerve stimulation. Vasoconstrictor responses to noradrenaline and histamine are antagonized by pimozide in a noncompetitive manner.     

Relation between adrenergic neurogenic contraction and alpha 1-adrenoceptor subtypes in dog mesenteric and carotid arteries and rabbit carotid arteries.

1. We examined the distribution of alpha 1-adrenoceptor subtypes and their relation to adrenergic neurogenic contraction induced by electrical transmural stimulation in the dog mesenteric and carotid arteries and the rabbit carotid artery. 2. In the dog mesenteric artery, contraction to noradrenaline was competitively inhibited by HV723 (pKB = 9.37) and prazosin (pKB = 8.40). Pretreatment with chlorethylclonidine (CEC) slightly attenuated only the contractions induced by low concentrations of noradrenaline. Contraction induced by electrical transmural stimulation was inhibited at lower concentrations of HV723 than of prazosin. 3. In the dog carotid artery, contraction to noradrenaline was inhibited with higher affinity by prazosin (pKB = 9.82) than by HV723 (pKB = 8.47). Prazosin was also more potent than HV723 in inhibiting the contraction to electrical stimulation. Pretreatment with CEC markedly attenuated or abolished contraction to noradrenaline and electrical stimulation. 4. In the rabbit carotid artery, prazosin inhibited noradrenaline-induced contraction biphasically (pKB = 9.91 and 8.60). After CEC pretreatment, contraction to noradrenaline was attenuated moderately and the high affinity site for prazosin was abolished. HV723 competitively inhibited the noradrenaline response with a similar pKB value (approximately 8.5) regardless of CEC treatment. Contraction to electrical stimulation was inhibited by prazosin more effectively than by HV723 in preparations not treated with CEC, while it was equipotently inhibited by both antagonists in CEC-treated preparations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prostacyclin mediates arachidonic acid-induced relaxation of rabbit isolated mesenteric arteries

Vasorelaxant effects of arachidonic acid have been attributed both to products of cyclooxygenase and alternative pathways. The present study tested whether the cyclooxygenase pathway functions to produce relaxation in mesenteric arterial rings and whether the arterial intimal layer is essential for these responses. Arachidonic acid dilated phenylephrine-contracted rings in a concentration-dependent manner. Abrasion of the intimal surface abolished the relaxant effect of arachidonic acid. The cyclooxygenase inhibitors--indomethacin, aspirin, and eicosatetraynoic acid--and a prostacyclin synthetase inhibitor, tranylcypromine, also significantly reduced or eliminated the vasodilator effect of arachidonic acid. Prostaglandins (PG) E1, E2, and I2 dilated precontracted rings whether the intimal layer was perturbed or not. Radioimmunoassayable concentrations of the stable PGI2 metabolite, 6 keto PGF1 alpha, were elevated following arachidonic acid addition in rings containing a functional endothelium. Cyclooxygenase inhibitors and tranylcypromine prevented the accumulation of 6 keto PGF1 alpha. The results are consistent with the hypotheses that arachidonic acid-induced relaxation of these rings is mediated by PGI2 and that the intimal layer is required for this conversion to PGI2.     

Effects of crotoxin on autonomic neuromuscular transmission in the guinea-pig myenteric plexus and vas deferens

A. Anadön and M. R. Martinez-Larran?aga. Effects of crotoxin on autonomic neuromuscular transmission in the guinea-pig myenteric plexus and vas deferens. Toxicon23, 963–972, 1985. — The effects of crotoxin, the neurotoxic complex from the venom of the South American rattlesnake Crotalus durissus terrificus on mammalian autonomic neuromuscular transmission, have been investigated. In the longitudinal muscle of the guinea-pig ileum, crotoxin induced a dose-dependent contraction which was followed by relaxation, in spite of the continued presence of the toxin. The contractile response was inhibited by indomethacin, tetrodotoxin, verapamil or nifedipine, but was unaffected by atropine, propranolol, mepyraroine or methysergide. In addition, crotoxin caused a presynaptic inhibition of the electrically-evoked twitch of the longitudinal muscle of the guinea-pig ileum. In the guinea-pig vas deferens crotoxin also caused an inhibition of the response to field stimulation. The inhibition was reversible after washing and the preparation remained insensitive to further doses of the toxin. The inhibitory effects of crotoxin were not mediated by noradrenaline and were not due to a non-specific smooth muscle depression, because it was not associated with any reduction in motor responses to acetylcholine, ATP, bradykinin or substance P. Pre-incubation of the guinea-pig vas deferens with indomethacin blocked the inhibitory effects of the toxin. This suggests that the presynaptic activity of crotoxin in the vas deferens might be mediated by prostaglandins.