5-Hydroxytryptamine-induced contractions of the human isolated saphenous vein: involvement of 5-HT2 and 5-HT1D-like receptors,and a comparison with grafted veins

Summary The receptors mediating the contractile effect of 5-hydroxytryptamine (5-HT) on the human isolated saphenous vein, obtained from 42 patients undergoing coronary bypass surgery, have been further characterized using a number of 5-HT-related drugs. The rank order of agonist potency was 5-carboxamidotryptamine (5-CT) 5-HT > methysergide sumatriptan -methyl-5-HT 5-methoxy-3-(1,2,3,6-tetrahydropyridin-4-yl)-1-Hindolesuccinate (RU 24969) 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI) > 2-methyl-5-HT > 8-hydroxy-2(di-n-propylamino)tetralin (8-OH-DPAT). Flesinoxan was inactive as an agonist. Ketanserin (1 mol/l) hardly affected sumatriptan-induced contractions but it caused a rightward shift of the upper part of the concentration-response curve of 5-HT and 5-CT. The same concentration of ketanserin caused a parallel rightward shift of the concentration-response curves of -methyl-5-HT and DOI with pK_B values of 7. 1 and 7.1, respectively. The responses to sumatriptan were antagonized by methiothepin (0.1 mol/l), metergoline (0.1 and 1 mol/l), rauwolscine (1 mol/l) and cyanopindolol (1 mol/l); the calculated pK_B values were 7.3, 6.9, 7.3, 6.7 and 6.5, respectively. Contractions to 5-HT were antagonized by methysergide (1 mol/l), methiothepin (0.1 mol/l; pKB = 7.1), ICS 205-930 (1 mol/l; pK_B = 5.9) and flesinoxan (30 mol/l; pK_B = 5.3). Remarkably, the contractions elicited by 2-methyl-5-HT were not attenuated by ICS 205-930, but were antagonized by methiothepin (0.1 mol/l) and, more markedly, by ketanserin (1 mol/l).There was a high correlation between the functional pD₂ values of 5-HT₁-like receptor agonists (5-CT, 5-HT, methysergide, sumatriptan, RU 24969 and 8-OH-DPAT) and their reported binding affinities for the 5-HT_1D receptor in human or calf brain membranes. Such a correlation for the antagonism of sumatriptan-induced responses was less marked than for the agonists, but of the 5-HT₁-like receptor subtypes it was the highest for the 5-HT_1D receptor identified in human or calf brain membranes.In 3 patients, undergoing heart transplantation, saphenous vein which had previously functioned as a graft for 6–11 years, was dissected out from the heart. Though the contractions to potassium were significantly smaller in the grafted veins, the pD₂ and E_max values (calculated as percentage of potassium-induced contractions) for 5-HT and sumatriptan were similar to those found in the veins obtained directly from the lower leg.It is concluded that contractions in the human isolated saphenous vein induced by 5-HT are mediated by 5-HT₂ receptors as well as by a 5-HT₁-like receptor resembling the 5-HT_1D subtype found in brain membranes. It is also to be noted that 2-methyl-5-HT, considered selective for the 5-HT₃ receptor, contracts the saphenous vein mainly via 5-HT₂ receptors.This study was supported by the Netherlands Heart Foundation, grant 89.252 Send offprint requests to W. A. Bax at the above address     

The role of brain norepinephrine in clonidine suppression of isolation-induced distress in the domestic chick

Distress vocalizations (DV) induced by social isolation were measured in 1-day-old domestic chicks after intracerebroventricular injections of drugs believed to exert their effects through the noradrenergic system. The -adreno-receptor agonist clonidine reliably suppressed DV rates at doses low as 0.08 g. When given alone, phentolamine, phenoxybenzamine, propranolol, sotalol, and yohimbine (adrenoreceptor antagonists) did not reliably after DV rates at doses that were not toxic. The clonidine DV suppression was reliably reversed by yohimbine (1.75 g), but by none of the other adrenoceptor antagonists or naloxone. 6-Hydroxydopamine at doses as high as 120 g, which essentially eliminated forebrain norepinephrine, failed to suppress DV rates reliably when given alone and, when given in combination with clonidine (0.1 g) or morphine (0.05 g), failed to reverse the severe DV suppression imposed by these drugs. The results suggest that clonidine suppresses DV rates in chicks through interaction with postsynaptic adrenoreceptors or by some means other than prejunctional ₂-adrenoreceptor stimulation.     

Antagonism by γ-aminobutyric acid of the stimulant effect of angiotensin II on cardiac sympathetic ganglia in spinal dogs

Summary The effects of angiotensin II and neuro-aminoacids administered through the right subclavian artery (i. a.) to the cardiac sympathetic ganglia were investigated in spinal dogs. Angiotensin II (1–8 g) elicited a dose-dependent positive chronotropic effect which was reduced after i. a. injection of saralasin (100g). The effect of angiotensin II was not reduced after combined treatment with either hexamethonium (10 mg/kg) plus atropine (0.1 mg/kg) or hemicholinium-3 (5 mg/kg) plus preganglionic stimulation. The dosedependent response to angiotensin II of heart rate was inhibited by GABA (50, 500g), GABOB (500g) and muscimol (50, 100g). The inhibition of the response to angiotensin II by a small dose of GABA (50g), but not by a high one (500g), was antagonized by i. a. injection of picrotoxin (2 mg). The positive chronotropism induced by bethanechol (25, 50g) and a small dose of acetylcholine (25g) were significantly inhibited by a high dose (500g) but not by a low dose (50g) of GABA. These results confirm that angiotensin II stimulates cardiac chronotropism by acting on the angiotensin II receptor located at the cardiac ganglia and show that this stimulant effect is antagonized by GABA.     

The depolarizing action of 5-hydroxytryptamine on rabbit isolated preganglionic cervical sympathetic nerves

Summary This study describes a depolarizing action of 5-hydroxytryptamine (5-HT) on rabbit isolated preganglionic cervical sympathetic nerves using an extracellular recording technique. From cumulative concentration-response curves for 5-HT (1 mol/1-1 mmol/1), the mean maximal depolarization was shown to be 277 ± 32 V and EC₅₀ was 9.4 mol/l(6.5–13.6 mol/l, geometric mean, 95% confidence limits, n = 42). The responses to 5-HT displayed marked tachyphylaxis. When cumulative concentration-response curves to 5-HT and 2-methyl-5-HT were determined in the same preparations (n = 4), the mean maximal response to 5-HT was 519 ± 167 V, EC₅₀ 32.2 mol/l (8.8–118 mol/l) and the mean maximal response to 2-methyl-5-HT was 317 ± 63 V, EC₅₀ 35.1 mol/l (12.9–95.5 mol/l, geometric means, 95 % confidence limits). The action of selective 5-HT antagonists was tested on repeated cumulative concentration-response curves to 5-HT. Neither methiothepin (0.1–1 mol/l, _n = 3) nor ketanserin (0.1–1 mol/l, n = 3) had an action on 5-HT responses. The selective 5-HT₃ antagonists MDL 72222, ICS 205-930 and SDZ 206–830 were all potent antagonists of the 5-HT depolarizations. The action of these antagonists was quantified by determining the apparent pA₂ from the dose ratios and a Schild plot. For MDL 72222 (1 nmol/1-0.1 mol/l), the apparent pA₂ was 9.1 ± 0.1 (n = 12), Schild plot: 9.2; for ICS 205–930 (0.1 nmol/l–3 nmol/1), the apparent pA₂ was 10.4 ± 0.1 (n = 11), Schild plot 10.3, and for SDZ 206–830 (0.03 nmol/l-1 nmol/1), the apparent pA₂ was 11.2 ± 0.1 (n = 12), Schild plot 11.2. 5-HT depolarizations were unaffected by hexamethonium (0.5 mmol/1). 5-HT depolarizations were reduced by superfusion with both Na-free (42 ± 8% of controls, n = 4) and Na/Ca-free media (35 ± 7% of controls, n = 4). It is concluded that 5-HT depolarizations of rabbit preganglionic cervical sympathetic nerve are mediated by 5-HT₃ receptors. The data with selective 5-HT₃ receptor antagonists suggest that the receptor profile may be more like that for the 5-HT₃ receptor on the terminals of sympathetic nerves than that for the 5-HT₃ receptor on the soma of superior cervical ganglion cells or on vagal afferent neurones. Send offprint requests to D. I. Wallis at the above address     

Effect of phorbol esters and polymyxin B on modulation of noradrenaline release in mouse atria

Summary Phorbol 12-myristate 13-acetate (PMA; 0.03, 0.1 and 1.0 mol/l), a protein kinase C activating phorbol ester, significantly enhanced the stimulation-induced (S-I) outflow of radioactivity at 5 Hz stimulation in mouse atria preincubated with [³H]-noradrenaline, whereas a phorbol ester which does not activate protein kinase C, phorbol 13-acetate (0.1 mol/l), had no effect. This suggests that protein kinase C may have a role in modulating sympathetic neurotransmission.Polymyxin B (7 and 21 mol/l), an inhibitor of protein kinase C, had no effect on the S-I outflow of radioactivity. However, it had a significant inhibitory effect in a concentration of 70 mol/l. Polymyxin B (21 mol/l) reduced the facilitation of the S-I outflow of radioactivity produced by PMA (0.03 mol/l), 8-bromo-cyclic AMP (90 mol/l), tetraethylammonium chloride (300 mol/l), and idazoxan (0.1 mol/l). Furthermore, when a higher frequency of stimulation was applied (10 Hz rather than 5 Hz), polymyxin B (21 pmol/1) by itself inhibited the S-I outflow of radioactivity.In the presence of a concentration of PMA (0.1 mol/l) that was maximally effective in enhancing the S-I outflow of radioactivity, both idazoxan (0.1 mol/l) and 8-bromocyclic AMP (90 mol/l) still enhanced the S-I outflow. This suggests that these agents are not operating through protein kinase C and further suggests that the inhibitory effect of polymyxin B on these agents cannot be due to inhibition of protein kinase C. The effects of clonidine on the S-I outflow were not affected by a maximally effective concentration of PMA (0.1 mol/l). These results suggest that protein kinase C is not involved in a ₂-adrenoceptor mediated modulation of noradrenaline release. Send offprint requests to I. F. Musgrave at the above address     

Dopamine inhibits prostaglandin F2α-induced depolarization of rabbit jejunal arteries via activation of DA1-receptors

Summary In rabbit jejunal arteries, the membrane potential of single smooth muscle cells decreased on the application of noradrenaline 3 mol/1. LY 171555 1 mol/1 did not change, whereas SKF 38393 10 mol/1 reversed the effect of noradrenaline. When prostaglandin F₂ (PGF₂) was used to evoke depolarization in the presence of prazosin 0.1 mol/1, rauwolscine 1 mol/1 and propranolol 1 mol/1, both SKF 38393 10 mol/1 and dopamine 10 mol/1 repolarized the membrane. SCH 23390 1 mol/1 antagonized the effects of SKF 38393 10 mol/1 and dopamine 10 mol/1. Thus, the change in membrane potential is mediated by a DA₁-recep-tor.     

Single-dose toxicokinetics of aluminum in the rat

The toxicokinetics of aluminum (Al) in male Wistar rats was studied after single intragastric (IG) doses of 1000 and 12000 g Al/kg and intravenous (IV) doses of 10, 100, 1000, and 12000 g Al/kg. Serial blood samples, daily samples of urine and feces as well as brain, liver, kidney, spleen, quadriceps muscle, and femur samples were collected. Al was measured by atomic absorption spectrometry. Al blood profiles after IV doses were adequately described by a two-compartment open model. Al toxicokinetics was dose dependent and appeared to plateau at 12000 g/kg. At IV doses between 10 and 1000 g/kg the terminal half-life of elimination from whole blood (t_1/2) increased from 29.9±7.8 to 209.3±32.6 min, and the total body clearance (CL) decreased from 2.45±0.64 to 0.28±0.03 ml min^–1 kg^–1. Following an IV bolus of 10 and 100 g/kg the administered Al was recovered completely from urine (94.4%±9.9% and 98.5%±3.2%). Twenty-nine days after the IV dose of 1000 g/kg daily renal excretion decreased to baseline values while only 55.1%±8.0% of the dose was excreted. Nineteen days after the single IV dose of 1000 g/kg Al accumulated in liver (28.1±7.7 versus 1.7±0.5 g/g of control rats) and spleen (72.5±21.1 versus <0.4 g/g). After the single 1000 g/kg IG dose no absorption of Al was detectable. The IG dose of 12000 g/kg resulted in a maximum blood Al level of 47.9±12.4 g/l after 50 min. The blood concentration time curve fitted a one-compartment open model with a half-life of absorption of 28.2±3.6 min and a t_1/2 of 81.2±20.2 min. Cumulative renal Al excretion was 0.18%±0.10% of the dose and oral bioavailability was 0.02%. Seventeen days after the 12000 g/kg IG dose the Al content in femur samples was increased (2.7±1.3 versus 0.6±0.4 g/g). In no case was fecal elimination of incorporated Al observed.     

Intrathecal substance P depresses the tail-flick response — Antagonism by naloxone

Summary Substance P injected into the lumbar subarachnoid space of rats depressed the tail-flick response to radiant heat in a dose-dependent way. The effective doses ranged from 0.1 g to 100 g per rat (ED 50: 1.5 g/rat). The maximum of the effect was reached 20 min after intrathecal injection and the effect lasted for about 30 min. An antinociceptive effect was also observed after intrathecal injection of substance P 1 g to spinal rats. The depression of the tail-flick response produced by intrathecal administration of substance P was abolished by intrathecal (5 g/rat) or i.p. (0.5 mg/kg) injections of naloxone.Supported by the Sonderforschungsbereich 38 Membranforschung     

Selective enhancement by an adenosine A1 receptor agonist of agents inducing contraction of the rat vas deferens

The adenosine analogue N⁶-cyclopentyladenosine (CPA), acting via postjunctional A₁ receptors, has been shown to enhance contractions of the rat vas deferens induced by adenosine 5-triphosphate (ATP), the sympathetic cotransmitter in this tissue. The aim of the present study was to examine the ability of CPA to enhance contractions induced by other contractile agents. CPA (0.01–0.3 M) enhanced contractions induced by exogenous ATP (10 M), 5-hydroxytryptamine (5-HT) (3 M), tyramine (10 M), 2-methyl-5-hydroxytryptamine (2-Me-5-HT) (10 M) and KCl (35 mM) and this enhancement was blocked by an A₁-selective concentration (3 nM) of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX). CPA failed to enhance contractions induced by exogenous noradrenaline (NA) (1 M or 10 M), bradykinin (0.1 M), phenylephrine (3 M) or carbachol (10 M).The contractions induced by ATP (10 M), 5-HT (3 M), 2-Me-5-HT (10 M) and KCl (35 mM) were unaffected by tetrodotoxin (1 M) as well as by desensitisation of the P_2x-purinoceptors with the ATP analogue adenosine 5-(, -methylene) triphosphonate. The contractions induced by tyramine (10 M) and 2-Me-5-HT (10 M) were blocked by prazosin (100 nM) or by imipramine (1 M). Ketanserin (10 nM) antagonised the response to 5-HT giving a dose-ratio of 12.9 corresponding to an apparent pA₂ of 9.1.In conclusion, the A₁-mediated effect was clearly selective for certain contractile agents and not due to a non-specific increase in contractility of the tissue. CPA enhanced contractions induced by both ATP and indirect sympathomimetics which release endogenous NA, and this enhancement of the two sympathetic cotransmitters may have a functional significance, and demonstrates the complexity of the neuromodulatory effects of adenosine in the rat vas deferens.     

Abolition by calcium antagonists of the autoregulation of renal blood flow

Summary Intra-arterial infusions of Ca antagonistic vasodilator substances such as nifedipine (1–10 g/min) and verapamil (30–100 g/min) inhibited renal autoregulation in the perfused dog kidney. Simultaneous infusion of 30 mg/min of CaCl₂ with nifedipine (3 g/min) or with verapamil (30 g/min) abolished the effects of these Ca antagonists. It is suggested that Ca ions play an important role in establishing renal autoregulation.     

Effects of GABA,dopamine, and substance P on the release of newly synthesized 3H-5-hydroxytryptamine from rat substantia nigra in vitro

Summary The effects of GABA, substance P and dopamine on the release of newly synthesized ³H-5-HT were investigated, using slices of rat substantia nigra superfused with l-³H-tryptophan in vitro. GABA (50 M) had no inhibitory effect on the potassium-evoked-release of ³H-5-HT. Substance P (50 M) and eledoisin (50 M) stimulated the spontaneous release of ³H-5-HT. This effect seems to be indirect and is possibly mediated by dopaminergic neurones, since the dopamine antagonist drug -flupenthixol (1 M) abolished the substance P-evoked release of 5-HT. Furthermore, it was found that substance P (10 M) stimulated ³H-dopamine release from nigral slices in vitro and the dopaminergic agonist apomorphine (50 M) also stimulated ³H-5-HT release. Substance P may, therefore, activate nigral dopaminergic neurones which then release dopamine from their dendrites. The release of dopamine may in turn stimulate 5-HT release from terminals of the raphe-nigral pathway.     

Release of ATP in rat vas deferens: origin and role of calcium

Release of endogenous ATP elicited by electrical (neural) stimulation and exogenous agonists was studied in the rat isolated vas deferens. The aims were to dissect neural and postjunctional contributions to the nerve activity-evoked overflow of ATP and to clarify the role of transmitter receptors and calcium in postjunctional ATP release.In tissues preincubated with [³H]-noradrenaline, electrical stimulation (100 pulses/10 Hz) elicited contraction and an overflow of tritium and ATP. Contractions as well as ATP overflow were reduced by prazosin 0.3 M and even more so by prazosin 0.3 M combined with suramin 300 M. They were also reduced by nifedipine 10 M and even more so by nifedipine 10 M combined with ryanodine 20 M (the additional effect of ryanodine on ATP overflow was not significant). In tissues not pretreated with [³H]-noradrenaline, exogenous noradrenaline 10 M and ,-methylene ATP 10 M elicited contraction and an overflow of ATP. Responses to noradrenaline were blocked by prazosin 0.3 M but not suramin 300 M and were greatly reduced by nifedipine 10 M and in Ca²⁺-free medium. Responses to ,-methylene ATP were blocked by suramin 300 M but not prazosin 0.3 M were reduced by nifedipine 10 M (effect on ATP overflow not significant) and were reduced even more in Ca²⁺-free medium. Neuropeptide Y 0.3 M caused only very small contraction and ATP overflow. The electrically as well as the agonist-evoked ATP overflow correlated well with the contraction responses except in experiments with suramin which retarded the removal, by vas deferens tissue, of ATP from the medium.Itsis concluded that the overflow of ATP from rat vas deferens elicited by electrical (neural) stimulation is at least 90% postjunctional, presumably smooth muscle, in origin. ATP is released from postjunctional cells as a consequence of both ₁-adrenoceptor and P₂-purinoceptor activation. Ca²⁺ is a second messenger in the postjunctional ATP release response; its major part enters through L-type channels. A purely neural overflow of ATP was not isolated under the conditions of the experiments. Correspondence to: R. Bültmann at the above address     

Neonatal exposure to zearalenone causes persistent anovulatory estrus in the rat

The effect of neonatal administration of zearalenone on the female reproductive system was studied in the rat. A single subcutaneous injection of 1.0 mg zearalenone to 3- or 5-day-old rats caused persistent vaginal estrus in adulthood. Ovaries in these animals contained many large follicles but no newly formed corpora lutea. The same effects were observed in rats which had received 100 g estradiol-17 in the neonatal period. Most rats which had received 100 g zearalenone or 10 g estradiol-17 showed regular 4-day estrous cycles and had newly formed corpora lutea in their ovaries. These results demonstrate that neonatal exposure to zearalenone produces persistent anovulatory estrus in the rat, the potency being about one tenth that of estradiol-17.     

Cardiovascular effects in rats of alpha1 and alpha2 adrenergic agents injected into the nucleus tractus solitarii

Summary The cardiovascular effects of selective alpha₁ and alpha₂ agonists and antagonists injected into the nucleus tractus solitarii (NTS) were studied in urethane-anesthetized rats. Methoxamine (0.3–3 g) injected bilaterally into the NTS caused a dose-dependent increase in blood pressure and heart rate. Phenylephrine (6 g) and an imidazolidine derivative St 587 (3 g) similarly injected also produced an increase in blood pressure, whereas a-methylnoradrenaline and an azepine derivative B-HT 920 (1 and 3 g) caused a decrease in blood pressure and heart rate. The pressor response to methoxamine (1 g) was markedly inhibited by prazosin (0.3 pg) injected into the same sites or hexamethionum (25 mg/kg, i. v.). Prazosin (0.3 g) alone injected bilaterally into the NTS did not affect the blood pressure, while yohimbine (0.1 g) similarly injected increased the pressure. These results suggest that in the rat NTS there exist alpha₁ adrenoceptors responsible for an increase in arterial pressure. The NTS alpha2 adrenoceptors seem to be involved in the tonic regulation of arterial pressure. Send offprint requests to T. Kubo at the above address     

Mechanisms of the contractile effects of flosequinoxan

In guinea-pig papillary muscles the positive inotropic effect of flosequinoxan (BTS) starting at 100 mol/1 amounted to 287.6 ± 34.2% at 300 mol/l without any effects on time to peak tension (103.9 ± 2%) and relaxation time (107.1 ± 6.7% of predrug value, respectively). 10 mol/l carbachol attenuated the positive inotropic effect of 300 mol/l to 166.5 ± 11.6% (n = 10). The phosphorylation state of the inhibitory subunit of troponin (TnI) and phospholamban(PLB) in [³²P]-labeled guinea-pig ventricular myocytes was increased starting at 100 mol/l amounting to 142.5 ± 12.6% and 130.9 ± 2.2% at 300 mol/l, respectively (n = 5). Furthermore, BTS (300 mol/l) decreased phosphorylase phosphatase activity by 23.1%. It is concluded that the contractile effects of BTS are accompanied by enhanced phosphorylation of regulatory proteins which could in part be due to inhibition of phosphorylase phosphatase activity.     

Metal concentrations in oldsquaw (Clangula hyemalis) during an outbreak of avian cholera, Chesapeake Bay, 1994

Forty out of 41 oldsquaw carcasses collected during a 3 month avian cholera outbreak in Chesapeake Bay, USA, in 1994 were culture positive for Pasteurella multocida. Pasteurella-positive birds collected in February had greater (p 0.05) mean (geometric) liver concentrations of cadmium (7.35 versus 3.71 g per g dry weight) and lower concentrations of selenium (9.90 versus 12.5 g per g dry weight) than Pasteurella-positive birds collected during March and April. The mercury content of the livers and cadmium content of the kidneys did not differ (p> 0.05) between birds collected early in the die-off and those collected in March and April. The liver and kidney concentrations of metals in the Pasteurella-positive birds collected in 1994 were compared to apparently healthy oldsquaw (n = 67) collected from Chesapeake Bay during 1985--1987, because healthy oldsquaw were not collected during the avian cholera outbreak in 1994. Compared to the apparently healthy oldsquaw collected in 1985--1987, the mean concentrations of cadmium (liver 4.32 versus 2.65 g per g dry weight and kidney 22.7 versus 11.5 g per g dry weight) were greater (p 0.05) in the oldsquaw which succumbed to avian cholera in 1994. In contrast, the liver concentrations of selenium (11.9 versus 17.8 g per g dry weight) and mercury (0.389 versus 1.83 g per g dry weight) were lower (p 0.05) in the birds from the 1994 die-off than for the apparently healthy oldsquaw collected in 1985--1987. Three birds from the 1985--1987 cohort and none of the birds from the 1994 cohort had liver lead concentrations greater than 4 g per g dry weight. The results of this study indicate a possible link between high cadmium tissue concentrations and susceptibility to avian cholera in oldsquaw     

Developmental Responses of a Terrestrial Insect Detritivore, <Emphasis Type="BoldItalic">Megaselia scalaris</Emphasis> (Loew) to Four Selenium Species

Megaselia scalaris (Loew) (Diptera: Phoridae) is an important and ubiquitous terrestrial detritivore that consumes both animal and plant material. Because both plants and animals convert selenium pollutants into various forms, the relative toxicities of ecologically relevant concentrations of sodium selenate, sodium selenite, seleno-L-methionine, and Se-(methyl) selenocysteine hydrochloride to larvae were assessed in diet bioassays. In addition, ovipositional preferences of adults and developmental effects on the eggs and larvae were measured. With chronic exposure selenocysteine was the most toxic of the selenium species to the larvae (LC₅₀: 83 g/g wet weight), followed by seleno-L-methionine (LC₅₀: 130 g/g), selenate (LC₅₀: 258 g/g), and selenite (LC₅₀: 392 g/g). Ovipositing females did not discriminate between the highest treatment concentrations of any of the pollutants as compared to the controls, indicating a lack of avoidance behavior. Larval development time was significantly increased with exposure to selenate at 100 g/g wet weight and above, selenite at 300 g/g and above, and at 50 g/g and 25 g/g and above for seleno-L-methionine and selenocysteine respectively. Pupal development was not affected by any of the selenium treatments. Significant differences between male and female adult eclosion times were observed, with females eclosing later than males as selenium concentrations increased. Significant decreases in larval survival relative to controls occurred at the lowest treatment tested (100 g/g) for both selenate and selenite and at 100 g/g for seleno-L-methionine, and 50 g/g for selenocysteine. The population level implications of lack of avoidance of contaminated food, and the effects of increased development times, reduced survivorship, and non-synchronized male and female emergence are discussed.     

Effects of (+)- and (±)-sotalol on repolarizing outward currents and pacemaker current in sheep cardiac Purkinje fibres

Summary This study was aimed to differentiate the action of (+)- and (±)-sotalol (10–1000 mol/l) on membrane currents which are active during the repolarization of cardiac action potentials Effects where studied in shortened sheep cardiac Purkinje fibres with the two-microelectrode voltage-clamp technique Action potentials were activated at a frequency of 0.25 Hz and membrane currents at 0.03 Hz or 0.05 Hz in most experiments.Out of the currents investigated the transient outward current (i_to) reacted most sensitively to (+)- and (±)-sotalol. I_to-amplitude was decreased on the average to 77% of reference at 10 mol/l and to 53% at 1000 mol/l (+)- or (±)-sotalol. The maximally available ito-current was decreased but the voltage-dependent control of inactivation was left nearly unchanged. The initial inwardly rectifying current (i_Ki), which propels the last repolarization phase of the action potential and controls resting potential to a large extent was reduced on the average to 93% of reference at 10 mol/l and to 62% at 1000 mol/l (+)- or (±)-sotalol. Time-dependent (delayed) outward current (i_K) was on the average not affected by (+)- or (±)-sotalol up to 100 mol/l and was decreased to 84% of reference current under the influence of 1000 mol/l. An initial outward current, which is activated at positive membrane potentials (i_inst) was not clearly affected by (+)- or (±)-sotalol at concentrations up to 1000 mol/l Pacemaker current (i_f) was not influenced by the drugs up to 100 mol/l. Only at 1000 mol/l was the amount of available i_f-current decreased to 79% of reference. (The potential-dependent control of activation was not affected) Time constants of time-dependent currents i_to, i_K and i_f did not change in concentrations up to 1000 mol/l of the drug.Action potential duration increased at (+)- or (±)-sotalol concentrations 10 mol/l and maximal prolongation was achieved at concentrations of 100–300 mol/l Resting potential remained nearly unchanged at these concentrations, but the membranes depolarized at 1000 mol/l. According to our data action potential prolongation in sheep Purkinje fibres under the influence of (+)- and (±)-sotalol correlates to the drug-induced block to i_to-current and inwardly rectifying i_K1-current.Supported by the Deutsche Forschungsgemeinschaft SFB 242, C 1 Send offprint requests to U. Borchard at the above address     

Analysis of the 5-HT receptor in rabbit saphenous vein examplifies the problems of using exclusion criteria for receptor classification

Summary 5-Hydroxytryptamine (5-HT) contracts ring preparations of rabbit saphenous vein via direct and indirect components, the latter being- compatible with a tyramine-like action at sympathetic nerve terminals. Here an attempt was made to establish the identity of the receptor mediating contraction directly, in terms of the currently accepted proposals (Bradley et al. 1986).Results with agonists suggested 5-HT₁-like receptor activation: methysergide behaved as a partial agonist with microcolar affinity and 5-HT effects were mimicked by 5-carboxamidotryptamine (5-CT) and GR43175. The agonist potency order was 5-CT > 5-HT > methysergide GR43175, the same as that reported at the 5-HT₁-like receptor in dog saphenous vein (Feniuk et al. 1985; Humphrey et al. 1988). Consistent with this, 5-HT effects were resistant to blockade by the selective 5-HT₃ receptor antagonist MDL72222 (1.0 mol/l). In contrast, methiothepin (0.01–0.3 mol/l), ketanserin (0.3–30.0 mol/l) and spiperone (0.3–30.0 mol/l) each produced surmountable antagonism which, although competitivv in nature only for methiothepin (pK_B = 9.45 ± 0.09, 17 d. f.), implied 5-HT₂ receptor involvement. The possibility that these discrepancies resulted from mixed populations of 5-HT₁-like and 5-HT₂ receptors can be excluded because; 1). Ketanserin and spiperone blocked the actions of 5-HT and the selective 5-HT₁-like receptor agonist GR43175 with equal facility and 2). Responses to all of the agonists studied were similarly antagonised by flesinoxan (pKB 6.4), a simple competitive antagonist at the receptor in rabbit saphenous vein. This novel result with flesinoxan demonstrates that the ligand displays affinity at 5-HT receptors other than the 5-HT_1A subtype.These data show that the 5-HT receptor in rabbit saphenous vein shares features in common with, and may be identical to, the 5-HT₁-like receptor in dog saphenous vein. However, unlike the latter it demonstrates qualities evident in both 5-HT₁-like and 5-HT₂ receptors and for this reason fails to meet the currently accepted criteria for admission into any of the recognised classes. It is suggested that this sort of problem reflects the generally unreliable behaviour of the available receptor antagonists and the emphasis which the Bradley et al. (1986) scheme places upon them for classification by exclusion. A complementary approach which provides a rigorous, quantitative basis for receptor differentiation uses finger-prints comprising affinity and relative efficacy estimates for a set of tryptamines. This study illustrates the power and economy of this approach by showing how affinity and relative efficacy finger-prints obtained using 5-HT, 5-CT, (±) -methyl-5-HT, 5-methyltryptamine and N,N-dimethyltryptamine establish a positive identity for the 5-HT receptor in rabbit saphenous vein and at the same time enable it to be distinguished from other 5-HT receptor types presently allocated to the 5-HT₁-like, 5-HT₂ and so-called orphan receptor classes.7Send offprint requests to G. R. Martin at the above address     

Effect of the phosphodiesterase inhibitor 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone (ZK 62711) on gastric secretion and gastric mucosal cyclic AMP

Summary The effect of the phosphodiesterase inhibitor 4-(3-cyclopentyloxy-4-methoxyphenyl)-2-pyrrolidone (ZK 62711) on gastric secretion and the cyclic AMP system of the gastric mucosa was studied in rats and guinea pigs. In rats, 0.03–0.3 moles/kg ZK 62711 i.v. stimulated acid and pepsin secretion in a dose-dependent manner and 0.03 moles/kg i.v. enhanced the effect of histamine. In guinea pigs no reproducible stimulation was found after intravenous injections of ZK 62711. The stimulation of gastric secretion in the rat by 0.3 moles/kg ZK 62711 i.v. was not affected by vagotomy but was totally inhibited by 10 moles/kg cimetidine i.v. The structurally related phosphodiesterase inhibitor, 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidine (Ro 20-1724), at the dose of 3.3 moles/kg i.v. stimulated gastric secretion in anaesthetised rats to a similar extent as 0.3 moles/kg ZK 62711 i.v. The content of cyclic AMP in the rat gastric mucosa in vivo was slightly increased by 10 moles/kg ZK 62711 i.v, whereas lower doses did not change it. Accumulation of cyclic AMP in the rat gastric mucosa by 50 moles/kg histamine i.v. was enhanced by simultaneous injections of 3.3 moles/kg ZK 62711 i.v. In rat gastric tissue slices in vitro 1–50 M ZK 62711 increased the level of cyclic AMP but 100 M histamine had no effect in the absence or presence of ZK 62711. In gastric mucosal slices of the guinea pig 10 and 50 M ZK 62711 increased the cyclic AMP content which effect was inhibited by 100 M cimetidine and enhanced the stimulatory effect of 100 M histamine on cyclic AMP. The activity of soluble cyclic AMP phosphodiesterase was inhibited by ZK 62711 in the rat (IC₅₀=18 M) and guinea pig gastric mucosa (IC₅₀=1.5 M). Adenylate cyclase was not affected in the homogenate of the guinea pig gastric mucosa. The results indicate that the phosphodiesterase inhibitor ZK 62711 which increases cyclic AMP levels in the gastric mucosa in vivo and in vitro, is a potent stimulator of gastric acid secretion.