Origin of 5-hydroxytryptamine-induced hyperpolarization of the rat superior cervical ganglion and vagus nerve

1 5-Hydroxytryptamine (5-HT)-induced membrane potential changes were recorded extracellularly from rat superior cervical ganglia (SCG) and cervical vagus nerves in vitro. 2 On the SCG, low concentrations of 5-HT (1 X 10(-8)-3 X 10(-7) M) induced concentration-related hyperpolarization responses. Higher concentrations of 5-HT (1 X 10(-6) 1 X 10(-4) M) induced complex responses which typically consisted of an initial hyperpolarization, followed by a depolarization and subsequent after-hyperpolarization. The depolarization, but not the initial hyperpolarization, was blocked by metoclopramide (3 X 10(-5) M), quipazine (1 X 10(-6) M) or MDL 72222 (1 X 10(-5) M). 3 5-HT-induced hyperpolarization of the SCG was potentiated when the amount of calcium chloride added to the superfusion medium was reduced from 2.5 to 0.15 mmol l-1. Hyperpolarization responses recorded from SCG preparations superfused with this low-calcium medium were unaffected by the substitution of lithium chloride for sodium chloride and were potentiated by the omission of potassium ions. Ouabain (1 X 10(-3) M) abolished both the hyperpolarization and the depolarization induced by 5-HT. 4 On the vagus nerve, 5-HT (1 X 10(-7) - 3 X 10(-5)M) did not induce initial hyperpolarization in either normal or low-calcium Krebs-Henseleit medium. However, in the latter solution only, depolarization responses induced by 5-HT at concentrations of 1 X 10(-6)M or greater were followed by hyperpolarization. Both the depolarization and the post-5-HT hyperpolarization were blocked by metoclopramide (3 X 10(-5)M) but were unaffected by spiperone (1 X 10(-7)M). 5 On the vagus nerve, post-5-HT hyperpolarization responses were selectively and reversibly inhibited by ouabain, and by superfusion with Krebs-Henseleit medium that was either potassium-free or contained lithium chloride in place of sodium chloride. 7 These results demonstrate the generation in the rat SCG of a 5-HT-induced hyperpolarization response that is not mediated through 5-HT3 receptors and is unlikely to be a consequence of depolarization. In contrast, on the rat vagus nerve, the post-5-HT hyperpolarization observed in the present study had the characteristics expected of depolarization-dependent activation of a sodium ion pump.     

Pharmacological characterization of angiotensin-induced depolarizations of rat superior cervical ganglion in vitro.

1. The depolarizing responses to angiotensin II and angiotensin III of the rat superior cervical ganglion have been characterized in vitro, by the use of peptidase inhibitors, peptide and non-peptide antagonists and dithiothreitol (DTT). 2. Angiotensin II and III depolarized the ganglion in a concentration-related manner. Angiotensin II was approximately 30 fold more potent than angiotensin III. 3. The endopeptidase inhibitor, bacitracin, increased the potency of angiotensin II and III by approximately 4 and 20 fold respectively. The aminopeptidase inhibitor, amastatin, further increased the potency of angiotensin III (but not angiotensin II) by approximately 4 fold. In the presence of bacitracin and amastatin, angiotensin II and III were equipotent. 4. The peptide antagonist [Ile7]angiotensin III (0.01-0.3 microM) produced a non-parallel rightward displacement of the angiotensin II concentration-response curve, with a suppression of the maximum response. The potency of [Ile7]angiotensin III was increased by bacitracin and amastatin. 5. The AT1-selective non-peptide antagonist losartan (DuP 753; 0.03 and 0.1 microM) produced a parallel rightward displacement of the angiotensin II concentration-response curve, with an apparent pKB of 8.3 +/- 0.1. A higher concentration of losartan (0.3 microM) depressed the maximum agonist response by 32 +/- 6.5%, possibly reflecting non-competitive behaviour of the antagonist. The potency of losartan was not influenced by bacitracin. 6. The AT2-selective non-peptide antagonist, PD123177 (3 microM) failed to antagonize the angiotensin II-induced depolarizations. 7. DTT (1 mM) produced a 22% reduction of the maximum response to angiotensin II.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pharmacological characterization of 5-hydroxytryptamine-induced depolarization of the rat isolated vagus nerve

A study has been made of the pharmacology of the 5-hydroxytryptamine (5-HT)-induced depolarization responses that can be recorded extracellularly from the rat isolated cervical vagus nerve. Phenylbiguanide (PBG) and 2-methyl-5-hydroxytryptamine (2-methyl-5-HT) were found to mimic the effects of 5-HT on the vagus nerve. Their EC50 values were respectively 2.0 fold and 3.9 fold greater than that of 5-HT. Metoclopramide behaved as a reversible competitive antagonist of depolarization induced by PBG and 2-methyl-5-HT, with pKB values of 6.48 +/- 0.04, respectively. These agreed well with the pKB value of 6.60 +/- 0.04 obtained previously for metoclopramide against 5-HT on the rat vagus nerve. 5-HT, PBG and 2-methyl-5-HT had no demonstrable agonist effects at non-5-HT receptors on the rat vagus nerve. Tropacaine and m-chlorophenylpiperazine were found to behave as reversible competitive antagonists of 5-HT-induced depolarization of the vagus nerve. The pKB values were 6.29 +/- 0.03 and 6.90 +/- 0.03, respectively. Quipazine, MDL 72222 and ICS 205-930 were also shown to be effective antagonists of 5-HT on the vagus nerve. However, although these compounds were highly potent, they all caused a marked concentration-dependent reduction in the amplitude of the maximum response to 5-HT. This behaviour was not consistent with a simple reversible competitive mechanism. The results are discussed with reference to the current classification of mammalian peripheral neuronal 5-HT receptors.     

Pharmacological characterization of 5-hydroxytryptamine-induced excitation of afferent cervical vagus nerve in anaesthetized rats.

1. An excitatory response to 5-hydroxytryptamine (5-HT) was measured from the afferent vagus nerve of anaesthetized rats. Measurements were determined by an extracellular recording from the whole nerve. 2. Intravenous bolus injection of 5-HT (1.56-100 micrograms kg-1) evoked a dose-dependent excitation of afferent vagus nerve activity. This response was blocked not only by a selective 5-HT3 receptor antagonist, GR38032F (10 and 100 micrograms kg-1), but also by a 5-HT2 receptor antagonist, ketanserin (10 and 100 micrograms kg-1). 3. Both a 5-HT3 receptor agonist, 2-methyl-5-HT (3.12-100 micrograms kg-1), and a 5-HT2 receptor agonist, alpha-methyl-5-HT (3.12-50 micrograms kg-1), produced a dose-dependent excitation of afferent vagus nerve activity. These excitatory effects were antagonized by GR38032F (10 micrograms kg-1) and ketanserin (10 micrograms kg-1), respectively. 4. A 5-HT1 like receptor agonist, 5-carboxamidotryptamine (50 micrograms kg-1), and a putative 5-HT4 receptor agonist, 5-methoxytryptamine (100 micrograms kg-1), failed to produce excitatory effects on the afferent vagus nerve. 5. These results suggest that the 5-HT-induced excitatory response of the afferent vagus nerve might be mediated not only via 5-HT3 receptors but also via 5-HT2 receptors in anaesthetized rats. It is unlikely, however, that either 5-HT1-like or putative 5-HT4 receptors are involved in the excitatory response of the afferent vagus nerve to 5-HT.     

Pharmacological differences between two muscarinic responses of the rat superior cervical ganglion in vitro.

1 Pharmacological differences have been observed between the muscarinic agonist-induced depolarizing and hyperpolarizing responses of the rat isolated superior cervical ganglion. 2 Pirenzepine (0.3 microM) selectively reduced the depolarizing response and unmasked the hyperpolarizing response. No such selectivity was observed with a concentration of N-methylatropine which was equipotent with pirenzepine in antagonizing the depolarizing response. 3 The neuromuscular blocking agents gallamine (10 microM) and pancuronium (3 microM) exhibited the oppositive selectivity to pirenzepine, both dramatically reduced the hyperpolarization but only slightly antagonized the depolarization. 4 The potencies of a range of agonists in evoking the depolarizing and hyperpolarizing responses, the latter in the presence of 0.3 microM pirenzepine, have been determined. Methylfurmethide failed to hyperpolarize the ganglion at concentrations which evoked maximal depolarizations. 5 The muscarinic hyperpolarization did not appear to be mediated by the secondary release of catecholamines. 6 It was concluded that the two muscarinic responses on the rat superior cervical ganglion, the slow depolarization and faster hyperpolarization, are mediated by different muscarinic receptor subtypes.     

Alpha-adrenergic receptors in the rat superior cervical ganglion.

α-Adrenergic receptor sites on membranes from rat superior cervical ganglia were measured by the specific binding of [³H]dihydroergocryptine, an α-adrenergic antagonist. The binding was specific, saturable, reversible, rapid, and stereoselective. Specific binding was displaced by α-adrenergic agonists in the expected order of potency and by α-adrenergic antagonists, but only weakly by a β-adrenergic agonist and antagonist. Decentralization of the ganglia by severing the preganglionic cholinergic nerve supply resulted in a marked decrease in the number of α-adrenergic receptors. The data suggest that about one-half the α-adrenergic receptors are located on the preganglionic cholinergic axon terminals and one-half on ganglionic cell membranes.     

Pharmacological characterization of 5-hydroxytryptamine-induced contraction in the chicken gastrointestinal tract

5-Hydroxytryptamine (5-HT) receptor subtypes involved in 5-HT-induced contraction of the chicken gastrointestinal tract were characterized pharmacologically using subtype-selective agonists and antagonists. The proventriculus (area of stomach adjacent to the oesophagus) and ileum are examined. 5-HT applied cumulatively caused sustained contraction of the proventriculus that was not decreased by tetrodotoxin, atropine or l-nitro-arginine methylester (l-NAME). alpha-Methyl-5-HT showed the same potency as that of 5-HT, indicating the involvement of the 5-HT(2) receptor. (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane (DOI), 5-methoxytryptamine and 1-(3-chlorophenyl)piperazine hydrochloride (mCPP) were potent, and 2-methyl-5-HT, 5-carboxamidotryptamine, BW723C86 and 6-chloro-2-(1-piperazinyl)pyrazine hydrochloride (MK212) were moderate, but (+/-)-8-hydroxy-2-dipropylaminotetralin hydrobromide (8-OH-DPAT), [endo-N-8-methyl-8-azabicyclo-(3,2,1)oct-3-yl]-2,3-dihydro-(1-methyl)ethyl-2-oxo-1H-benzimidazol-1-carboxamide (BIMU-8) and cisapride were weak agonists. Correlation of pEC(50) values of these agonists with documented pEC(50) values for 5-HT(2C) receptor was higher than 5-HT(2A) and 5-HT(2B). Cinanserin, ketanserin, methiothepin, methysergide, mianserin, (8-[5-(2,4-dimethoxy-5-(4-trifluoromethylphenylsulphonamido)phenyl-5-oxopentyl)-1,3,8-triazaspiro[4,5]decane-2,4-dione hydrochloride (RS102221), N-(1-methyl-1H-indolyl-5-yl)-N'-(3-methyl-5-isothiazolyl)urea (SB204741), spiperone and N-desmethylclozapine concentration-dependently inhibited the contractile responses to 5-HT. Correlation of pK(b)/pA(2) of antagonists with documented pK(i) for 5-HT(2C) receptor was highest among 5-HT(2) receptor subtypes. In the methysergide- and ketanserin-treated proventriculus, 5-HT, 2-methyl-5-HT and cisapride did not enhance the electrical field stimulation (5 Hz)-induced cholinergic contractions. 5-HT applied non-cumulatively caused transient contraction of ileum, and the responses were partly decreased by atropine or tetrodotoxin. 5-Methoxytryptamine, alpha-methyl-5-HT, 5-carboxamidotryptamine, L692,247 and DOI were potent agonists. However, 2-methyl-5-HT, cisapride, BW723C86, 8-OH-DPAT and 5-nonyloxytryptamine, mCPP and MK212 were less effective. Ketanserin and methysergide decreased the 5-HT-induced ileal contraction, but neither GR113808 nor SB269970 inhibited the responses. In conclusion, 5-HT causes contraction of the proventriculus via 5-HT(2C)-like receptors present on smooth muscle. 5-HT also causes contraction of the ileum, but the underlying mechanisms are complex, involving neural and smooth muscle components, and both 5-HT(1)- and 5-HT(2)-like receptors. Neural 5-HT receptors similar to 5-HT(3)/5-HT(4) receptors were not found in the chicken proventriculus and ileum.     

The antagonist actions of WAY 100135 and its enantiomers on 5-HT1A receptor-mediated hyperpolarization of the rat isolated superior cervical ganglion

Summary A grease-gap extracellular recording technique was used to detect 5-HT_1A receptor-mediated hyper-polarizing responses to 5-hydroxytryptamine (5-HT) in the rat isolated superior cervical ganglion. In the presence of the novel 5-HT_1A receptor antagonist, WAY 100135 [N-tert-butyl-3-(4-(2-methoxyphenyl)piperazin-1-yl)-2-phenylpropanamide], the responses to 5-HT were antagonised in a competitive manner with a pA₂ value of 7.2 (6.9–8.5) and Schild plot slope of 1.0 (0.4–1.6), n = 20. The antagonist activity was greater in the (+) than the (–)enantiomer of WAY-100135. The pA₂ value of the (+)enantiomer was 7.5 (7.2–8.0), Schild plot slope 1.2 (0.8–1.6), n = 17. In contrast the (–)enantiomer had weak antagonist activity (pA₂ 6.3±0.25, n = 3). No agonist activity of WAY-100135 or its enantiomers were observed in this study. Correspondence to K. F. Rhodes at the above address     

Characterization of the adenosine receptors of the rat superior cervical ganglion.

1. Adenosine analogues caused hyperpolarization and inhibition of the depolarizing response to muscarine of the rat isolated superior cervical ganglion (SCG) measured by a 'grease gap' recording technique. The receptors mediating these responses have been characterized by use of a range of selective adenosine analogues and adenosine receptor antagonists. 2. In decreasing order of potency N6-cyclopentyladenosine (CPA), 2-chloroadenosine (2CA), adenosine, 2-phenylaminoadenosine (PAA), caused concentration-dependent hyperpolarizations whilst N6-(9-fluorenylmethyl)adenosine (PD 117,413) was inactive at up to 100 microM. 3. The order of potency of adenosine analogues in depressing depolarization caused by a submaximal concentration of muscarine (100 nM) was: CPA > R-PIA = 2CA > NECA > S-PIA > BZA > adenosine > PAA, where R- and S-PIA = R(-)- and S(+)-N6-(2-phenylisopropyl)adenosine, NECA = 5'N-ethylcarboxamidoadenosine and BZA = N6-benzyladenosine. PD 117,413 was inactive at concentrations up to 100 microM. The maximum inhibitions of the muscarine-induced depolarization by CPA, 2CA, NECA and BZA were similar. R-PIA, S-PIA and PAA produced similar maximal inhibitions which were significantly smaller than those produced by CPA. 4. Hyperpolarizations caused by adenosine were antagonized by the P1-purinoceptor selective antagonist 1,3-dimethyl-8-phenylxanthine (8PT) and by the selective A1-adenosine receptor antagonist, 1,3-dipropyl-8-(4-((2-aminoethyl)amino)carbonylmethyloxyphenyl++ +)xanthine (XAC). Hyperpolarizations caused by CPA, adenosine and PAA were antagonized by the A1-selective antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) but not by the A2-selective antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX).(ABSTRACT TRUNCATED AT 250 WORDS)     

5-Hydroxytryptamine evokes three distinct responses on the rat superior cervical ganglion in vitro

In addition to the 5-HT3-mediated fast depolarisation, 5-hydroxytryptamine (5-HT) evoked two additional responses on the rat superior cervical ganglion: a hyperpolarisation and a slow depolarisation. These responses appeared to be direct actions on 5-HT receptors since they were present in a low calcium medium containing tetrodotoxin and were not abolished by a variety of non-serotonin antagonists. The hyperpolarisation was not antagonised by 5-HT3 or 5-HT2 antagonists. The 5-HT1 ligands 5-carboxamidotryptamine (5-CT) and 8-OH-DPAT also evoked a hyperpolarisation. The hyperpolarisation was antagonised by six 5-HT1A antagonists including WB-4101 and spiroxatrine. It was therefore concluded to be mediated by a 5-HT1A receptor. The slow depolarisation was only evoked by 5-HT. The receptor involved in this response, however, could not be determined. We conclude that in addition to 5-HT3 receptors the rat superior cervical ganglion possesses 5-HT1A receptors and another uncharacterised 5-HT receptor.     

On the histamine-induced depolarization of the isolated superior cervical ganglion of the rat.

1. Using a grease-gap technique, we studied the action of histamine on the d.c. potential recorded between the internal carotid nerve and the main body of the isolated superior cervical ganglion of the rat. 2. A small, slow depolarization was evoked by 10-300 microM histamine. This response was not reduced by lowering the calcium concentration in the superfusing medium (from 2.5 to 0.1 mM), or by superfusing tetrodotoxin, N-methylatropine, or propranolol (all at 1 microM). 3. Mepyramine (10 nM) antagonized this depolarization, but cimetidine (10 microM), metiamide (30 microM), burimamide (10 microM) and impromidine (1 microM) did not. Two other agonists also evoked a mepyramine-sensitive slow depolarization. The rank order of potencies was histamine greater than N alpha-methyl-histamine greater than 2-methyl-histamine. 4. At concentrations greater than 1 mM, histamine also evoked a larger, faster depolarization. This response was undiminished by reducing the calcium concentration of the medium to 0.1 mM or by adding 1 microM tetrodotoxin. The rank order of potency for the agonists was N alpha-methyl-histamine greater than histamine approximately 2-methyl-histamine. The histamine-induced fast response was not antagonized by any of the above-mentioned antagonists. It was slightly reduced by (+)-tubocurarine (100 microM) and N-methylbicuculline (100 microM) but such effects were not consistent with the blockade of nicotinic or GABAA receptor-mediated responses. 5. It was concluded that histamine depolarized the isolated superior cervical ganglion of the rat by activating H1 receptors. Relatively high concentrations of histamine also evoked a fast depolarization of this preparation, but this did not appear to be mediated by H1, H2 or H3 receptors.     

Action of purine and pyrimidine nucleotides on the rat superior cervical ganglion.

1. Using a grease-gap technique, we have investigated the effects of purine and pyrimidine nucleotides on the d.c. potential of the rat isolated superior cervical ganglion (SCG). 2. Of the purines tested, adenosine, adenosine 5'-triphosphate (ATP), beta,gamma-methylene-adenosine 5'-triphosphate (beta,gamma-MeATP) at up to 300 microM produced concentration-dependent hyperpolarizations, whereas 2-methyl-thio-ATP (2-Me.S.ATP) and alpha,beta-methylene-ATP (alpha,beta-MeATP) depolarized ganglia. Of the pyrimidines tested, uridine 5'-triphosphate (UTP) produced concentration-dependent depolarizations and cytosine 5'-triphosphate (CTP) at 1000 microM produced considerably smaller but significant depolarizations. In contrast uridine 5'-monophosphate (UMP) at 1000 microM hyperpolarized ganglia. The relative order of potency of purines and pyrimidines to depolarize ganglia was: UTP > alpha,beta-MeATP >> CTP > 2-Me.S.ATP and to hyperpolarize ganglia was: adenosine = beta,gamma-MeATP > ATP > UMP. 3. The ability of purines and pyrimidines to alter the depolarizing response caused by muscarine and of purines to alter depolarization induced by gamma-aminobutyric acid (GABA) was determined. The relative order of potency of nucleotides in depressing submaximal depolarization caused by muscarine (100 nM) was: adenosine = ATP > beta,gamma-MeATP whereas 2-Me.S.ATP, alpha,beta-MeATP and UTP did not significantly alter depolarization caused by muscarine. At 100 microM beta,gamma-MeATP and adenosine but not ATP potentiated GABA-induced depolarizations. 4. Hyperpolarizations caused by adenosine, ATP, beta,gamma-MeATP and UMP and depolarizations caused by alpha,beta-MeATP were enhanced in medium containing reduced concentrations of calcium (0.1 mM) and potassium (2 mM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of 5-hydroxytryptamine uptake on the apparent 5-hydroxytryptamine antagonist potency of metoclopramide in the rat isolated superior cervical ganglion.

Metoclopramide, 1 X 10(-6) -1 X 10(-4) M, was found to behave as a reversible, competitive antagonist of 5-hydroxytryptamine (5-HT)-induced depolarization of the rat isolated vagus nerve (VN) and superior cervical ganglion (SCG). The pKB values were 6.60 (+/- 0.04) and 5.74 (+/- 0.07), respectively. The possibility that this apparent difference in potency was due to saturable 5-HT uptake was investigated. The SCG, but not the VN, accumulated tritium-labelled 5-HT via a saturable, sodium- and temperature-dependent mechanism. Ganglionic 5-HT uptake was blocked by desmethylimipramine (IC50 1.4 X 10(-6)M), chlorimipramine (8.7 X 10(-9) M), zimelidine (1.5 X 10(-7) M), paroxetine (4.3 X 10(-8) M) and citalopram (6.2 X 10(-8) M). The 5-HT uptake inhibitor paroxetine, 1 X 10(-6) M, did not modify the apparent 5-HT antagonist potency of metoclopramide on the VN, but raised the pKB obtained against 5-HT on the SCG from 5.74 (+/- 0.07) to 6.25 (+/- 0.03). It is suggested that the observed difference in the potency of metoclopramide as a 5-HT antagonist on the rat VN and SCG was due to saturable 5-HT uptake in the latter preparation. The results do not support a difference in the 5-HT receptors mediating depolarization on the VN and SCG.     

Structure-activity relationship of a pyrimidine receptor in the rat isolated superior cervical ganglion.

1. The effects of pyrimidines and purines on the d.c. potential of the rat isolated superior cervical ganglion (SCG) have been examined by a grease-gap technique to determine the structure-activity requirements of the receptor activated by pyrimidines, i.e. a pyrimidinoceptor. 2. 5-Aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl (ZTP), the pyrimidines, cytidine 5'-triphosphate (CTP), uridine 5'-triphosphate (UTP) and thymidine 5'-triphosphate (TTP) and the purines, adenosine 5'-triphosphate (ATP; in the presence of an A1-purinoceptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX) (1 microM)), adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), guanosine 5'-triphosphate (GTP), inosine 5'-triphosphate (1TP) depolarized ganglia in a concentration-dependent manner. The relative order of ZTP and purine 5'-triphosphates in depolarizing ganglia was ZTP > or = ATP gamma S > > ATP > or = ITP = GTP, and for the pyrimidine 5'-triphosphates UTP > TTP > or = CTP. Depolarizations evoked by ATP gamma S were followed by concentration-dependent hyperpolarizations at 100 and 1000 microM. 3. At concentrations of between 0.1 microM and 1 mM, uridine 5'-diphosphate (UDP), uridine 5'-diphosphoglucose (UDPG) and uridine 5'-diphosphoglucuronic acid (UDPGA) evoked significant and concentration-dependent depolarizations, whereas uridine 5'-monophosphate (UMP), uridine and uracil were inactive or produced small (< 45 microV) depolarizations. The relative order of potency of uridine analogues in depolarizing ganglia was UDP > or = UTP > UDPG > UDPGA > > uracil > or = UMP = pseudouridine > or = uridine. At 3 and 10 mM, uridine produced concentration-dependent hyperpolarizations. Nikkomycin Z, a nucleoside resembling UTP (viz. the triphosphate chain at the 5'-position on the ribose moiety being replaced by a peptide), was inactive between 1 microM and 1 mM. Generally, a concentration of 10 mM was required before thymidine, 6-azathymine, 6-azauracil or 6-azauridine depolarized ganglia. 4. Suramin (300 microM), a P2-purinoceptor antagonist, significantly depressed depolarizations evoked by alpha, beta-methylene-ATP (alpha, beta-MeATP; 100 microM), ATP gamma S (100 microM), CTP (1 mM), GTP (1 mM), ZTP (30 microM) and ATP (300 microM) in the presence of DPCPX (1 microM). Suramin reversed a small depolarization evoked by UMP (1 mM) into a small hyperpolarization. In contrast depolarizations evoked by UDP, UTP, UDPG (all at 100 microM) and TTP (300 microM) were unaltered or enhanced by suramin. 5. It is concluded that the rat SCG contains distinct nucleotide receptors including a P2-purinoceptor (activated by alpha, beta-MeATP, ATP, GTP, ITP and ZTP) and a pyrimidinoceptor (activated by UTP, UDP, UDPG, UDPGA and TTP). The pyrimidinoceptor on rat SCG neurones had specific structure activity requirements with the di- and triphosphates of uridine being the most effective depolarizing agonists examined.     

Biphasic dose-response curve to muscarine on the rat superior cervical ganglion

The dose-response curve for the muscarine-induced depolarisation of the rat isolated superior cervical ganglion, studied over the concentration range of 3 nM-1 mM, was biphasic. An apparent maximum was obtained at around 1-3 microM muscarine, but this was only a plateau between the two parts of the curve. Two cardioselective antagonists, gallamine (10 microM) and AF-DX 116 (1 microM) had a complex action on this dose-response curve. The dose-response curve between 0.01 and 0.3 microM was shifted to the right, the responses around 3 microM muscarine were enhanced, but the dose-response curve over 30 microM muscarine was unaffected. The M1-selective antagonist pirenzepine (0.05 microM) depressed all parts of the dose-response curve, but it still appeared biphasic. Pretreatment of the ganglion with pertussis toxin (1 microgram/ml) enhanced the depolarisation to muscarine 0.01-1000 microM and the dose-response curve became less biphasic. Like gallamine and AF-DX 116, pertussis toxin abolished the muscarinic M2-mediated hyperpolarisation of the ganglion recorded in 0.3 microM pirenzepine. It is concluded that the presence of an underlying M2-mediated hyperpolarisation contributes to the biphasic nature of the dose-response curve to muscarine.     

Action of surugatoxin on nicotinic receptors in the superior cervical ganglion of the rat.

Surugatoxin (SGTX, 0.1-2 muM) reversibly depressed orthodromic transmission and antagonized the depolarizing action of carbachol on the isolated superior cervical ganglion of the rat. The apparent dissociation equilibrium constant against carbachol-induced depolarization (measured in the presence of hyoscine) was 58 and 76 nM determined at 0.2 and 2 muM respectively. SGTX (2muM) did not reduce the depolarizing effects of (+/-)-muscarine, gamma-aminobutyric acid or angiotensin, but did reduce that to 5-hydroxytryptamine. Release of [3H]-acetylcholine following repetitive (10 Hz) preganglionic sympathetic stimulation was maintained in the presence of 2 muM SGTX. It is concluded that SGTX has a high and selective affinity for ganglionic nicotinic receptors.     

Membrane potential changes induced by 5-hydroxytryptamine in the rabbit superior cervical ganglion.

1. Changes in resting membrane potential induced by 5-hydroxytryptamine (5-HT) have been measured in the excised ganglion by the sucrose-gap technique. 2. 5-HT produced a rapid depolarization, the threshold concentration for depolarization being around 10 muM. With concentrations of 100 muM or greater, repolarization began during the course of the superfusion; this was followed by prolonged tachyphylaxis. 3. Tachyphylaxis was largely avoided by making injections into the superfusion stream. Standard injections of 0.2 mumol 5-HT dissolved in 0.2 ml of Krebs solution were used routinely and could be given at 20-30 min intervals to evoke relatively constant responses. 4. The response to an injection consisted of a rapid depolarization, followed by a rapid repolarization and subsequent after-hyperpolarization. The threshold quantity for depolarization was around 0.01 mumol, while the ED50 estimated from 6 dose-response curves was 0.12 +/- 0.02 mumol (mean +/- s.e. mean). 5. Injections of 5-HT (0.2 mumol), choline (10 mumol) and acetylcholine (9.9 mumol) produced depolarizations of similar magnitude. 6. Monoamine oxidase inhibitors failed to alter substantially the amplitude of depolarizations to 5-HT. 7. 5-HT depolarizations were unaltered in amplitude when the impermeant anion benzenesulphonate was substituted for the chloride ion in Krebs solution, but were initially markedly reduced in amplitude in a sodium-deficient medium; some recovery of the response subsequently occurred. The depolarization which persisted in sodium-deficient solutions was much reduced or abolished when calcium ions were then removed from the superfusion medium. Removal of either calcium ions alone or potassium ions from the superfusion fluid did not reduce depolarization amplitude. 8. The after-hyperpolarization was abolished in sodium-deficient solutions, usually increased in potassium-free solutions, reduced or abolished by ouabain or nicotine, but unaffected by calcium free solutions. 9. A depolarizing action of 5-HT on presynaptic terminals in the ganglion appears probable.