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D I Wallis

Publications and source records attributed to D I Wallis.

At least 19 recordsLinked to original sources

Properties of putative cardiac and non-cardiac neurones in the rat stellate ganglion.

Intracellular recordings were made from isolated left or right stellate ganglia of Wistar rats and the morphology of neurones studied after intracellular injection of hexammine cobaltic chloride or back-filling from the post-ganglionic nerve with cobalt lysine complex. The experiments attempted to identify the location, electrophysiological properties, morphology and chemosensitivity of putative cardiac neurones in the ganglion. These were identified by antidromic activation of the axon in a cardiac nerve and compared with neurones projecting towards the brachial plexus (non-cardiac neurones). Putative cardiac neurones were localized in the ganglion around the postganglionic nerve entry zone and showed considerable morphological diversity. They had complex dendritic trees with, on average, seven dendrites. They included both phasic and tonic neurones and were depolarized by muscarinic agonists, angiotensin and substance P; they invariably had a synaptic input from the sympathetic trunk and from a T1 or T2 ramus and, in 16% of cells, from a cardiac nerve. Non-cardiac neurones were more widely scattered through the stellate ganglion but were not clearly different in morphology, resting membrane potential or the proportion of phasic and tonic cells from putative cardiac neurones. They also showed depolarizing responses to muscarinic agonists, angiotensin and substance P. Angiotensin responses of stellate ganglion cells were blocked by the peptide antagonist, saralasin (1 microM).

Action Potentials

FAST and SLOW ipsilateral and contralateral spinal reflexes in the neonate rat are modulated by 5-HT.

1. Three ipsilateral (MSR, PSR, IPSI SLOW) and two contralateral segmental reflexes (CON FAST, CON SLOW) were recorded from L4 or L5 ventral roots of the neonate rat spinal cord in vitro. MSR, PSR and CON FAST were evoked from lower threshold afferents; more intense stimulation evoked IPSI SLOW and CON SLOW. 2. Kainate/AMPA receptors were involved in mediation of MSR, PSR, CON FAST, IPSI SLOW and CON SLOW and NMDA receptors in mediation of CON FAST, IPSI SLOW and CON SLOW. 3. All five reflexes were depressed by 5-HT (IC50 1.2-7.9 microM; order of sensitivity, CON SLOW > CON FAST = IPSI SLOW > MSR = PSR); and by 5-CT (IC50 1.9-8.8 nM; order of sensitivity, MSR > IPSI SLOW = CON FAST = CON SLOW > PSR). alpha-Me-5-HT also depressed all five reflexes. 4. Dipropyl-5-CT selectively depressed MSR and CON SLOW (IC50 90-170 nM) but was less potent than 5-CT. 8-OH-DPAT selectively depressed MSR (IC50 1.1 microM), IPSI SLOW and CON SLOW (IC50 5.7-7.6 microM), while methylsergide depressed only MSR (IC50 26 nM). 5. Phenyl biguanide and m-chlorophenyl biguanide (5-HT3 receptor agonists) had no significant effects on any reflex. 6. It is concluded that a 5-HT1-like receptor mediates depression of the MSR. A different receptor or a mixed population of receptors, but not 5-HT3 receptors, mediate inhibition of PSR, CON FAST, IPSI SLOW and CON SLOW.

Animals

Serotonin and L-norepinephrine as mediators of altered excitability in neonatal rat motoneurons studied in vitro.

The actions of serotonin on the membrane properties of motoneurons and on the synaptic responses evoked by stimulating the segmental dorsal root have been investigated using intracellular recording in a neonatal rat hemisected spinal cord preparation in vitro. Superfusion with serotonin produced concentration-dependent depolarizations (EC50 32.1 microM) with an apparent increase in input resistance and increase in motoneuron excitability. During serotonin depolarizations an increase in membrane noise was seen. At higher serotonin concentrations repetitive firing was induced. Sensitivity to serotonin was enhanced by blockade of neuronal uptake with citalopram, when the EC50 was 1.4 microM. The depolarization was mimicked by alpha-methyl-5-hydroxytryptamine (EC50 11.7 microM). Serotonin depolarizations were blocked by ketanserin (0.1 and 1 microM), ritanserin (1 microM), spiperone (0.1 and 1 microM) and LY 53857 (1 microM). A norepinephrine-induced depolarization of motoneurons, which was mimicked by L-phenylephrine and antagonized by prazosin, is probably mediated by an alpha 1-adrenoceptor. An inhibitory action of serotonin was also apparent. The frequency and amplitude of spontaneous postsynaptic potentials and the response following dorsal root stimulation were markedly reduced. This action was mimicked by 5-carboxamidotryptamine and 8-hydroxy-2-(n-dipropylamino)tetralin, but was not antagonized by ketanserin (1 microM), ritanserin (1 microM), methiothepin (1 microM), metergoline (1 microM), spiperone (1-10 microM) or 21-009 (1-10 microM). It is proposed that the depolarization and increase in excitability of spinal motoneurons is mediated by a serotonin (5-HT2) receptor subtype.

Animals

Further studies on the action of 5-hydroxytryptamine on lumbar motoneurones in the rat isolated spinal cord.

Using the hemisected spinal cord of the neonate rat, the effects of altered external Ca, thyrotrophin-releasing hormone (TRH) and a number of antagonists were tested on depolarizations evoked by 5-hydroxytryptamine (5-HT). Responses of populations of motoneurones were recorded via a ventral root. 5-Hydroxytryptamine depolarizations were not Ca-dependent but were enhanced in amplitude in Ca-free solutions. Raised Mg reversed this enhancement. 5-Hydroxytryptamine depolarizations persisted in the presence of Mn (1.5-3 mmol/l). TRH depolarized motoneurones; there was no evidence of modulation of 5-HT responses on concurrent application of TRH. Ritanserin (0.1 mumol/l) had a modest blocking action on 5-hydroxytryptamine depolarizations reducing the maximum; 1 mumol/l ritanserin caused a greater antagonism which was unsurmountable (pIC50 5.2). Ritanserin (0.1 or 1 mumol/l) did not depress responses to noradrenaline (NA). Ketanserin (0.1 mumol/l) caused a blockade of slow onset, equilibrium with the receptors requiring 1 h. Blockade by 0.01, 0.1 and 1 mumol/l ketanserin was concentration-dependent (pIC50 6.2). Ketanserin 1 mumol/l, but not at lower concentrations, depressed noradrenaline responses. Mianserin (0.1 mumol/l) also caused a blockade of slow onset; 0.1 or 1 mumol/l produced a flattening of the 5-hydroxytryptamine concentration-response curve but did not depress noradrenaline responses (pIC50 4.7). The pIC50 for spiperone was 8.0. DOI (10-100 mumol/l) had no detectable agonist action but at concentrations of 0.01 and 0.1 mumol/l it acted as an antagonist. Equilibration with the receptors occurred over 2 h. DOI (0.01 mumol/l) depressed 5-hydroxytryptamine but not noradrenaline responses; higher concentrations of DOI also depressed noradrenaline responses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Inhibition of reflex responses of neonate rat lumbar spinal cord by 5-hydroxytryptamine.

1. Monosynaptic (MSR) and polysynaptic (PSR) segmental reflex responses were recorded from a ventral root of the neonate rat hemisected spinal cord. Amplitudes of the two components were monitored with a peak height detector. 2. 5-Hydroxytryptamine (5-HT) depressed the MSR and PSR in a concentration-dependent manner. The IC50 for MSR depression was 9.5 +/- 3.2 microM and for PSR depression was 9.0 +/- 4.8 microM. 3. Blockade of neuronal uptake of 5-HT by citalopram (0.1 microM) greatly increased sensitivity to 5-HT. In the presence of citalopram, the IC50 for MSR depression was 30 +/- 18 nM and for PSR depression was 89 +/- 23 nM. 4. 5-HT did not depress the MSR or the PSR by releasing glycine since strychnine (1 microM) did not prevent these actions of 5-HT. 5. 5-Carboxamidotryptamine (5-CT), 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), RU 24969, 1-[3-(trifluoromethyl)phenyl]-piperazine (TFMPP) and methysergide were full agonists for depression of the MSR. The IC50 for 5-CT was 3.6 +/- 0.5 nM, for 8-OH-DPAT was 0.4 +/- 0.04 microM, for TFMPP was 0.93 +/- 0.3 microM and for methysergide was 21.8 +/- 3.0 nM. The order of potency was 5-CT greater than methysergide greater than 5-HT greater than 8-OH-DPAT greater than TFMPP. 6. 8-OH-DPAT, RU 24969, TFMPP and methysergide had either no or only a minor action in reducing the PSR. 5-CT caused a 50% depression at the highest concentration tested (30 nM). 7. Neither ketanserin (1 microM) nor spiperone (1 microM) caused appreciable blockade of 5-HT depression of the MSR or 5-HT depression of the PSR. 8. Blockers of neuronal 5-HT uptake (citalopram 0.1 or 1 microM, fluvoxamine 1 microM) usually reduced the MSR and, to a lesser extent, the PSR. Reflex depressions were reversed by ketanserin (1 microM). 9. It was concluded that 5-HT has a potent depressant action on segmental reflexes; depression of the MSR is unrelated to depolarization of motoneurones. Although depression of the MSR was mimicked by 5-HTIA receptor ligands, the action of endogenous 5-HT may be mediated through 5-HT2 receptors. Exogenous 5-HT may act at a mixture of 5-HT receptor subtypes to depress the MSR.

Animals

Analysis of the actions of 5-hydroxytryptamine on the rabbit isolated vagus nerve.

Depolarization and reduction in the C fibre compound action potential (C spike) in response to 5-HT were recorded simultaneously from rabbit isolated vagus nerve. 5-HT (0.1-100 mumol/l) was applied either as single concentrations or cumulatively and EC50 and IC50 values measured from individual concentration-response curves. The EC50 values for depolarization (cumulative curves: 2.33, 1.64-3.33 mumol/l, geometric means and 95% confidence limits, n = 31; non-cumulative curves: 3.99, 2.89-5.52 mumol/l, n = 9) were significantly higher than IC50 values for C spike reduction (cumulative curves: 1.25, 0.91-1.74 mumol/l, n = 30; non-cumulative curves: 1.41, 0.72-2.76 mumol/l, n = 8). Complex effects on the C spike were observed, suggesting a susceptible group of C fibres and a 5-HT-resistant component to the C fibre action potential. The motor nerve C fibres in the vagus nerve appear insensitive to 5-HT, whereas the sensory C fibres were sensitive to 5-HT. Phenylbiguanide had a similar selective effect on the C spike, while the depolarizing agents, 1,1-dimethyl-4-phenylpiperazinium (DMPP) and gamma-aminobutyric acid (GABA) did not. Cumulative concentration-response curves for depolarization and C spike reduction could be repeated reproducibly if an interval of 90 min was left between determinations. Up to 6 curves could be generated from one preparation. The 5-HT uptake inhibitor, citalopram (0.1 and 1 mumol/l), had no effect on cumulative concentration-response curves.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

Antagonism of the effects of 5-hydroxytryptamine on the rabbit isolated vagus nerve by BRL 43694 and metoclopramide.

Depolarization and reduction in the C fibre compound action potential (C spike) in response to 5-HT were recorded simultaneously from rabbit isolated vagus nerve. 5-HT (0.1-100 mumol/l) was applied cumulatively and EC50 and IC50 values measured from individual concentration-response curves. Blockade of 5-HT responses by the 3-indazole carboxamide, BRL 43694, was investigated and compared with the blocking action of metoclopramide. BRL 43694 was a selective antagonist of 5-HT responses. A concentration of 10 nmol/l BRL 43694, which nearly abolished the depolarization and reduction of the C spike evoked by 5-HT (100 mumol/l), had no effect on similar responses evoked by DMPP (100 mumol/l) or GABA (100 mumol/l). Blockade of 5-HT responses by BRL 43694 (0.3 nmol/l) was slow in onset, a plateau blockade occurring after equilibrium of tissue with antagonist for 2 to 3 h. Metoclopramide induced a blockade of rapid onset. The maximal blockade was apparent within 30 min of application. Full recovery in the responsiveness of the tissue to 5-HT was observed within 30 min of washing out metoclopramide. BRL 43694 at concentrations of 0.3, 1, 3 and 10 nmol/l caused a progressive rightward shift of the concentration-response curves to 5-HT. At the highest concentration of antagonist, there was some depression of the maximal 5-HT response. The apparent pA2 estimated from the Schild equation was 10.03 +/- 0.09 (mean +/- SEM, n = 20) against 5-HT depolarization and 10.31 +/- 0.1 against C spike reduction. Schild plots had slopes not significantly different from 1.0.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials

Involvement of alpha 1-adrenoceptors in the depolarizing but not the hyperpolarizing responses of motorneurones in the neonate rat to noradrenaline.

Superfusion of the hemisected lumbar spinal cord in the neonate rat with solutions containing 10(-6) to 10(-3) M noradrenaline (NA), elicited graded depolarizations recorded from ventral roots, with a mean EC50 value of 15.1 +/- 1.5 microM (mean +/- SEM, n = 37). Repeated concentration-response curves to NA could be determined from the same preparation. Adrenaline had a similar depolarizing action (EC50 9.9 +/- 1.7 microM, n = 11). Blockade of neuronal uptake of NA by desipramine (2 x 10(-6) M) caused some potentiation of submaximal responses to NA and shifted the EC50 to 6.0 +/- 1.7 microM (mean +/- SEM, n = 14). The depolarizing response to NA was unaffected by DL-propranolol (10(-7) M) or yohimbine (10(-7) M). Prazosin (5 x 10(-9), 10(-8) and 10(-7) M) reduced the responses and caused a progressive rightward shift of the concentration-response curve. The onset of blockade by prazosin was slow, superfusion for at least 90-120 min being required before the blockade plateaued. Prazosin (5 x 10(-9) and 10(-8) M) caused a surmountable blockade, the apparent pA2 being 8.3 +/- 0.2 (mean +/- SEM, N = 9). Depolarizations induced by NA were also antagonised by phentolamine (10(-6) M). An initial hyperpolarizing response to NA was unmasked after exposure to prazosin in 90% of preparations and was associated with a reduction in the spontaneous activity of the motorneurones. Both the hyperpolarization and reduction in spontaneous activity were attenuated by yohimbine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

5-Hydroxytryptamine depolarizes neonatal rat motorneurones through a receptor unrelated to an identified binding site.

Superfusion of hemisected lumbar spinal cord of the neonatal rat with solutions containing 10(-6) to 10(-3) M 5-hydroxytryptamine (5-HT) elicited depolarizations of graded amplitude which were recorded from motorneurons through a ventral root. Maximum responses (amplitude 1.0 +/- 0.1 mV, mean +/- SEM, n = 30) were evoked by 10(-4) M 5-HT. Repeated concentration-response curves could be determined from the same preparation. There was no involvement of 5-HT2 receptors in the depolarizing response to 5-HT, since neither ritanserin nor ICI 169, 369 showed any antagonist action. Amongst agents with activity at 5-HT1A sites, the selective 5-HT1A receptor agonist, 8-hydyroxy-2-(di-N-propylamino) tetralin (8-OH-DPAT), neither mimicked the action of 5-HT nor antagonised it, while spiperone (10(-8)-10(-7 M) antagonised responses to 5-HT in a concentration-related manner. Responses to 10(-4) M noradrenaline, used as a control depolarizing agent, were unaffected by spiperone. The onset of blockade by spiperone was slow, 1 hr being required for equilibration of the tissue with antagonist. The blockade was surmountable by larger concentrations of 5-HT. Concentration-response curves to 5-HT were shifted to the right in an approximately parallel manner by spiperone. The dose ratios measured from these curves at the EC50 level, yielded an apparent pA2 of 8.24 +/- 0.14 (mean +/- SEM, n = 15), although the Schild plot of the data had a slope less than unity. The lack of activity of the selective 5-HT1B receptor agonist, RU 24969, and the 5-HT1B receptor antagonists, (+/-) cyanopindolol and quipazine, indicated that 5-HT1B receptors were not involved in the 5-HT response of motorneurones to 5-HT. Mesulergine, metergoline and cyproheptadine also antagonised responses of motorneurones to 5-HT, producing a surmountable blockade. Mesulergine (10(-8), 3 x 10(-8) and 10(-7) M caused a progressive rightward shift of the concentration-response curves, but 10(-7) M depressed the maximum response to 5-HT. Responses to noradrenaline were not affected by these concentrations of mesulergine. The apparent pA2 for blockade of 5-HT responses by mesulergine, calculated from experiments in which there was a parallel displacement of the concentration-response curves, was 8.75 +/- 0.11 (mean +/- SEM, n = 10).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

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

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 mumol/l-1 mmol/l), the mean maximal depolarization was shown to be 277 +/- 32 microV and EC50 was 9.4 mumol/l (6.5-13.6 mumol/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 microV, EC50 32.2 mumol/l (8.8-118 mumol/l) and the mean maximal response to 2-methyl-5-HT was 317 +/- 63 microV, EC50 35.1 mumol/l (12.9-95.5 mumol/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 mumol/l, n = 3) nor ketanserin (0.1-1 mumol/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.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

A comparison of fast and slow depolarizations evoked by 5-HT in guinea-pig coeliac ganglion cells in vitro.

1. 5-Hydroxytryptamine (5-HT) was applied by pressure ejection to coeliac ganglion cells of the guinea-pig maintained in vitro and responses measured intracellularly. 2. Cells responded in one of three ways to 5-HT: by (a) a fast, transient depolarization (43%), (b) a fast transient followed by a slow depolarization (biphasic response, 30%) or (c) a slow sustained depolarization (25%). 3. Fast depolarizations (response (a) above] were graded according to the duration of the ejection pulse. Maximal responses had a mean amplitude of 12 +/- 0.8 mV, a duration of 6.4 +/- 1.0 s, a latency of 0.4 +/- 0.1 s, were associated with a fall in membrane input resistance, increased in amplitude by hyperpolarization and probably mediated by an increased conductance to Na and K. The estimated reversal potential was -22.8 +/- 2.4 mV (n = 14). The maximal fast response seen in biphasically-responding cells (b) appeared similar to fast response (a). 4. Fast depolarizations (a) showed marked tachyphylaxis and were abolished by superfusion of the ganglion with 5-HT (100 microM). They were reduced in amplitude by tubocurarine (10-100 microM, pIC50 4.4), MDL 72222 (1-5 microM, pIC50 5.8), quipazine (1 microM reduced responses by 65 +/- 15%, n = 3), ICS 205-930 (1 microM reduced responses by 64 +/- 14%, n = 7) and metoclopramide (10 microM reduced responses by about 45%), but were unafected by methysergide (up to 1 microM) or hexamethonium (up to 1 mM). 5. Slow depolarizations (c) varied in amplitude with the duration of the ejection pulse. Maximal responses had a mean amplitude of 6.4 +/- 0.7 mV, a duration of 62 +/- 6 s, a latency of 3.5 +/- 0.8 s and were reduced in amplitude by methysergide (0.1-1 microM, pIC50 6.5) but not by MDL 72222 (1 microM). The maximal slow component in biphasically-responding cells (b) was similar in amplitude and duration to slow response (c), was partially blocked by methysergide (1-5 microM) in 4 of 6 cells and was enhanced by tubocurarine (50 microM) which reduced the fast component. 6. Slow depolarizations (b,c) were associated with either a small reduction or no change in membrane input resistance depending on the cell studied. Hyperpolarization had variable effects on slow depolarization amplitude. 7. It was concluded that the fast, phasic depolarization is mediated by an ionic mechanism and by receptors both of which are distinct from those involved in the slow depolarization. The receptor mediating the fast depolarization is a 5-HT3 receptor while that mediating the slow depolarization has yet to be identified.

Animals

Responses to 5-hydroxytryptamine evoked in the hemisected spinal cord of the neonate rat.

1. Superfusion of isolated hemisected spinal cord from neonate rats with 5-hydroxytryptamine (5-HT) (10(-6) to 10(-3) M) evoked concentration-related depolarizations. The maximal depolarization elicited by a concentration of 10(-4) M was 1.0 +/- 0.1 mV (mean +/- s.e.mean, n = 30). Noradrenaline in a similar range of concentrations also elicited depolarizations. 2. The depolarizations probably originate in motoneurones as a result of direct interaction of the amines with these cells, since responses were unaltered by tetrodotoxin (10(-7) M) or Ca2+-free/Mg2+-rich medium. 3. 5-Carboxamidotryptamine (5-CT), S(+)-alpha-methyl-5-hydroxytryptamine (alpha-Me5-HT) and 5-methoxytryptamine (5-MeOT) evoked similar depolarizations to 5-HT. Tryptamine evoked depolarizations of smaller maximal amplitude. 5-Hydroxytryptophan, 2-methyl-5-hydroxytryptamine, 8-hydroxy-2-(di-N-propylamino) tetralin hydrobromide (8-OH-DPAT) and 5-methoxy-3-[1,2,3,6-tetrahydro-4-pyridinyl]-1-H-indole succinate (RU 24969) had no depolarizing action. 4. Concentration-response (CR) curves were determined for 5-HT, 5-CT, alpha-Me5-HT, 5-MeOT and tryptamine. The ED50 value for 5-HT was 20.5 +/- 1.2 microM. The equipotent molar ratios (EPMRs) for 5-CT and alpha-Me5-HT were close to unity, while 5-MeOT was approximately 3 times and tryptamine 13 to 14 times less potent than 5-HT. 5. The relative agonist potency of 5-HT with respect to other tryptamine analogues capable of depolarizing motoneurones was increased when 5-HT uptake was blocked by citalopram (10(-7) M). In the presence of citalopram, 5-HT was 2.7 times more potent than alpha-Me5-HT and 16.9 times more potent than 5-CT. The apparent order of potency was 5-HT greater than alpha-Me5-HT greater than 5-CT (greater than 5-MeOT much greater than tryptamine). 6. The monoamine oxidase inhibitor, pargyline (5 x 10(-4) M), had no effect on depolarizations to 5-HT, 5-CT or alpha-Me5-HT. 7. Methiothepin, 1 alpha H, 3 alpha, 5H-tropan-3-yl-3,5-dichlorobenzoate methanesulphonate (MDL 72222) and [3 alpha-tropanyl]-1H-indole-3-carboxylic acid ester hydrochloride (ICS 205-930) had no effect on 5-HT depolarizations elicited in motoneurones. Ketanserin (0.75 x 10(-7) M to 10(-6) M) showed modest antagonistic action and depressed maximal response amplitude; the pIC50 was 6.5. 8. Methysergide (10-8 to 10- 7M) was a potent antagonist of responses to 5-HT. CR curves were displaced to the right and flattened in the presence of the antagonist. The pIC5o assessed from the effect on depolarizations evoked by 5-HT 1O-4M was 7.5. 9. It is concluded that 5-HT acts directly to depolarize mammalian spinal motoneurones through receptors that are also activated by 5-CT, alpha-MeS-HT and 5-MeOT and are blocked by methysergide. The receptor profile, although not 5-HT3-like, does not clearly coincide with that for either 5-HT1-like or 5-HT2 receptors.

Animals

An electrophysiological study of ganglion blockade by paraquat and diquat.

1. The bipyridilium herbicides, particularly paraquat, have chemical and toxicological features in common with the bi-quaternary ammonium ganglion blockers. 2. Paraquat and diquat were tested for ganglion blocking activity. Rabbit cervical ganglia were superfused with both agents and subsequently with hexamethonium to confirm susceptibility to ganglion blockade. 3. No evidence for ganglion blockade was found at either supra maximal or sub maximal stimulation, and none following repetitive stimulation. 4. The similarities and differences between the bipyridyl herbicides and bi-quaternary ganglion blockers can be explained on the basis of their structures. 5. It is concluded that neither paraquat nor diquat have significant ganglion blocking activity.

Animals

Further studies on the blockade of 5-HT depolarizations of rabbit vagal afferent and sympathetic ganglion cells by MDL 72222 and other antagonists.

The blocking action of MDL 72222 (1 alpha H, 3 alpha, 5 alpha H-tropan-3-yl-3, 5-dichlorobenzoate at 5-hydroxytryptamine (5-HT) receptors on nodose (NG) and superior cervical ganglia (SCG) has been investigated further. The sucrose-gap technique was used to record potential changes from populations of neurones. The surmountable blockade induced by small concentrations of the antagonist was quantified and the blocking potency compared with that of a number of other compounds. In nodose ganglia three 4-5 point dose-response (DR) curves were established, using bolus injections of 5-HT (5-80 nmol). The mean amplitude of the response to 80 nmol was 4.18 +/- 0.53 mV and the ED50 was 18.2 nmol. Second and 3rd dose-response curves showed small displacements to the right, indicating a slight reduction in sensitivity. In superior cervical ganglia responsiveness was less. Amounts of 5-HT ranging from 20 to 320 nmol evoked dose-related depolarizations. The mean amplitude of the response evoked by 320 nmol 5-HT was 1.7 +/- 0.14 mV. Three 4-5 point dose-response curves could be elicited from a single ganglion. The ED50 was 55.8 nmol. Initial, 2nd and 3rd dose-response curves could be superimposed, there being no significant rightward shift. The results confirm that MDL 72222 is a potent, selective antagonist at 5-HT receptors in nodose and superior cervical ganglia. In the nodose ganglion, after equilibration for 1 hr with 10(-8) or 10(-7) M MDL 72222, dose-response curves for 5-HT showed rightward, parallel shifts. In contrast, 10(-6) M MDL 72222 or prolonged exposure (3-4 hr) to 10(-8), 10(-7) or 10(-6) M caused larger rightward shifts of the dose-response curves and depressed the maximum responses. In the superior cervical ganglion, equilibration for 1 hr with concentrations of 10(-8) or 10(-7) M produced effects on the dose-response curves similar to those seen in the nodose ganglion, but longer exposures (3-4 hr) did not depress the maximum. Apparent pA2 values were determined from individual experiments on both the nodose and superior cervical ganglia, where MDL 72222 (10(-7) M or less, for 1 hr) caused parallel or near parallel shifts of dose-response curves. In the nodose ganglion the apparent pA2 was 7.7 +/- 0.1, while in the superior cervical ganglion it was 7.8 +/- 0.1 (mean +/- SEM). The nature of the blockade induced by prolonged exposures or by concentrations greater than 10(-7) M is discussed.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials

Fast and slow depolarizing responses of guinea-pig coeliac ganglion cells to 5-hydroxytryptamine.

Responses to 5-hydroxytryptamine (5-HT) applied by pressure ejection to guinea-pig coeliac ganglion cells were recorded intracellularly in vitro. Three types of responses were observed: a fast depolarization (43% of cells), a slow depolarization (25%) and a biphasic response comprising an initial fast followed by a slow depolarization (30%). Fast depolarizations had a mean duration of 6.4 s and were associated with a decrease in membrane input resistance. On hyperpolarizing the membrane, responses increased in amplitude; the mean reversal potential was -23 mV. Fast responses were nearly eliminated in a Na-free medium but were unaffected by low Ca/high Mg solution or by low Cl medium. Fast responses were depressed by MDL 72222, quipazine, metoclopramide, and by relatively high concentrations of D-tubocurarine, but unaffected by methysergide. Slow responses had a mean duration of 62 s and were associated either with no change or a slight fall in membrane resistance. hyperpolarizing the membrane, slow responses were increased in most but not all cells tested. Slow depolarizations were depressed by methysergide but unaffected by agents that suppressed the fast responses. The biphasic response displayed electrophysiological and pharmacological characteristics of both the fast and slow depolarizations. It is concluded that guinea-pig coeliac ganglion cells display either a rapid depolarization, a slow depolarization or a biphasic response to 5-HT, that the fast depolarization appears to be mediated by a conductance increase to Na and K, while a different mechanism appears to underlie the slow depolarization, and that the two responses are mediated by different kinds of 5-HT receptor.

Animals

The depolarizing action of 5-hydroxytryptamine on rabbit vagal afferent and sympathetic neurones in vitro and its selective blockade by ICS 205-930.

Depolarizing responses to 5-hydroxytryptamine (5-HT) were recorded from rabbit nodose (NG) and superior cervical (SCG) ganglia using the sucrose-gap technique. The antagonist potency and selectivity of ICS 205-930 ([3 alpha-tropanyl]-1H-indole-3-carboxylic acid ester) were investigated. In NG, 5-HT (5 to 80 nmol) evoked depolarizations of graded amplitude. The ED50 was 18.2 (10.9-30.5) nmol (geometric mean, 95% confidence limits). Responses were blocked surmountably by ICS 205-930, 10(-11) and 10(-10) M, the threshold for blockade being below 10(-11) M. Parallel, rightward shifts in dose-response curves were seen with these concentrations of antagonist, but at higher concentrations (10(-9) and 10(-8) M) there was a further rightward shift with reduction in slope and maximum of the curves. In SCG, where 5-HT (20 to 320 nmol) evoked depolarizations of graded amplitude and the ED50 was 55.8 (22.3-139.6) nmol (geometric mean, 95% confidence limits), ICS 205-930 had a similar inhibitory effect to that observed in NG. The apparent pA2 values for the surmountable blockade produced by ICS 205-930 at concentrations of 10(-11) and 10(-10) M were 10.2 +/- 0.2 for NG and 10.4 +/- 0.1 for SCG (means +/- s.e. mean). ICS 205-930 was selective in its action since it had no effect on dimethylphenylpiperazinium (DMPP) responses in either ganglion or on GABA responses in NG. This study provides quantitative evidence on the blocking action of ICS 205-930 at neuronal 5-HT receptors using a technique that allows the depolarizing responses evoked by the amine to be directly recorded.

Animals

The depolarizing action of 5-hydroxytryptamine on rabbit vagal primary afferent and sympathetic neurones and its selective blockade by MDL 72222.

MDL 72222 (1 alpha H,3 alpha,5 alpha H-tropan-3-yl-3,5-dichlorobenzoate) is a novel compound with potent and selective blocking actions at certain excitatory 5-hydroxytryptamine (5-HT) receptors on mammalian peripheral neurones. In the present study, the sucrose-gap technique has been used to record depolarizing responses to 5-HT from the cells of the rabbit nodose and superior cervical ganglia and to investigate the potency and selectivity of MDL 72222 as an antagonist of these responses. On nodose ganglia, responses to 5-HT were inhibited surmountably by MDL 72222 at concentrations up to 100 nmol/l. The threshold for antagonism was 2-10 nmol/l and the apparent pA2 value (Schild 1947) was 7.7 +/- 0.2, n = 10. Blockade was selective since responses to GABA and noradrenaline were unaffected by MDL 72222, 100 nmol/l. With concentrations of MDL 72222 higher than 100 nmol/l, antagonism was concentration-related but not in a manner consistent with simple competitive antagonism and even a concentration of 1 mumol/l failed to abolish the response to 5-HT. The results from the superior cervical ganglion were essentially similar to those obtained from the nodose ganglion. The threshold concentration of MDL 72222 for inhibition of 5-HT was 1-10 nmol/l and blockade was selective in that depolarizing responses to dimethylphenyl-piperazinium (DMPP) was unaffected by a concentration of MDL 72222 of 1 mumol/l. The data provide direct evidence that MDL 72222 is a potent and selective antagonist of the receptors for 5-HT which mediate depolarizing responses in vagal primary afferent cell bodies and in sympathetic ganglion cells.

Animals

Properties of visceral primary afferent neurons in the nodose ganglion of the rabbit.

The active and passive membrane properties of rabbit nodose ganglion cells and their responsiveness to depolarizing agents have been examined in vitro. Neurons with an axonal conduction velocity of less than 3 m/s were classified as C-cells and the remainder as A-cells. Mean axonal conduction velocities of A- and C-cells were 16.4 m/s and 0.99 m/s, respectively. A-cells had action potentials of brief duration (1.16 ms), high rate of rise (385 V/s), an overshoot of 23 mV, and relatively high spike following frequency (SFF). C-cells typically had action potentials with a "humped" configuration (duration 2.51 ms), lower rate of rise (255 V/s), an overshoot of 28.6 mV, an after potential of longer duration than A-cells, and relatively low SFF. Eight of 15 A-cells whose axons conducted at less than 10 m/s had action potentials of longer duration with a humped configuration; these were termed Ah-cells. They formed about 10% of cells whose axons conducted above 2.5 m/s. The soma action potential of A-cells was blocked by tetrodotoxin (TTX), but that of 6/11 C-cells was unaffected by TTX. Typically, A-cells showed strong delayed (outward) rectification on passage of depolarizing current through the soma membrane and time-dependent (inward) rectification on inward current passage. Input resistance was thus highly sensitive to membrane potential close to rest. In C-cells, delayed rectification was not marked, and slight time-dependent rectification occurred in only 3 of 25 cells; I/V curves were normally linear over the range: resting potential to 40 mV more negative. Data on Ah-cells were incomplete, but in our sample of eight cells time-dependent rectification was absent or mild. C-cells had a higher input resistance and a higher neuronal capacitance than A-cells. In a proportion of A-cells, RN was low at resting potential (5 M omega) but increased as the membrane was hyperpolarized by a few millivolts. A-cells were depolarized by GABA but were normally unaffected by 5-HT or DMPP. C-cells were depolarized by GABA in a similar manner to A-cells but also responded strongly to 5-HT; 53/66 gave a depolarizing response, and 3/66, a hyperpolarizing response. Of C-cells, 75% gave a depolarizing response to DMPP.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals