Non-adrenergic Non-cholinergic Inhibitory Junction Potential Research Articles

Involvement of purinergic nerves in the NANC inhibitory junction potentials in pigeon oesophageal smooth muscle

1. Electrical field stimulation (EFS) (0.5 ms in train of 2-32 Hz for 300 ms) in smooth muscle of pigeon oesophagus, in the presence of atropine (1 microm) and guanethidine (1 microm), elicited an inhibitory response consisting of a transient hyperpolarization (inhibitory junction potential, IJP) associated with muscle relaxation. 2. Sodium nitroprusside (SNP, 100 microm) induced hyperpolarization correlated to mechanical relaxation. 3. The nitric oxide (NO) synthase inhibitor N(omega)-nitro-l-arginine (from 0.1 to 100 microm) caused a concentration-dependent reduction of electromechanical response to EFS indicating a role for NO in this response. 4. Apamin (1 microm) reduced both IJP and relaxation to EFS but was without effect on the response to SNP indicating a role for purines, which are also blocked by apamin. 5. Adenosine, AMP, ADP and ATP (all from 1 microm to 1 mm) application caused transient hyperpolarization and muscular relaxation with the following order of potency: adenosine > AMP > ADP > ATP. 6. Inhibitory responses evoked by purines are TTX (1 microm) insensitive but they were inhibited by apamin. This indicates that a purine component for the non-adrenergic non-cholinergic (NANC) response exists but the purine receptor site is not located on the neurone. 7. Overall these results suggest that NANC inhibitory response elicited by EFS presents two different components apamin-sensitive, probably purines-mediated and apamin-insensitive probably NO-mediated as apamin only partially block the response to EFS.

Effect of exogenous ATP on canine jejunal smooth muscle.

Intracellular recordings were made from the circular smooth muscle cells of the canine jejunum to study the effect of exogenous ATP and to compare the ATP response to the nonadrenergic, noncholinergic (NANC) inhibitory junction potential (IJP) evoked by electrical field stimulation (EFS). Under NANC conditions, exogenous ATP evoked a transient hyperpolarization (6.5 +/- 0.6 mV) and EFS evoked a NANC IJP (17 +/- 0.4 mV). Omega-conotoxin GVIA (100 nM) and a low-Ca(2+), high-Mg(2+) solution abolished the NANC IJP but had no effect on the ATP-evoked hyperpolarization. The ATP-evoked hyperpolarization and the NANC IJP were abolished by apamin (1 microM) and N(G)-nitro-L-arginine (100 microM). Oxyhemoglobin (5 microM) partially (38.8 +/- 5.5%) reduced the amplitude of the NANC IJP but had no effect on the ATP-evoked hyperpolarization. Neither the NANC IJP nor the ATP-evoked hyperpolarization was affected by P2 receptor antagonists or agonists, including suramin, reactive blue 2, 1-(N, O-bis-[5-isoquinolinesulfonyl]-N-methyl-L-tyrosyl)-4-phenylpiperazine , pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid, alpha, beta-methylene ATP, 2-methylthioadenosine 5'-triphosphate tetrasodium salt, and adenosine 5'-O-2-thiodiphosphate. The data suggest that ATP evoked an apamin-sensitive hyperpolarization in circular smooth muscle cells of the canine jejunum via local production of NO in a postsynaptic target cell.

Apamin- and nitric oxide-sensitive biphasic non-adrenergic non-cholinergic inhibitory junction potentials in the rat anococcygeus muscle.

1. Changes in membrane potential following electrical field stimulation (EFS; 1, 2 and 5 pulses at 5 Hz, 0.5 ms duration, 60-80 V) of non-adrenergic non-cholinergic (NANC) inhibitory nerves in the rat isolated anococcygeus muscle were measured using standard intracellular recording techniques. Resting membrane potential ranged between -60 and -70 mV. 2. In the presence of guanethidine (30 microM), atropine (1 microM), propranolol (1 microM) and phentolamine (0.05 microM) to establish NANC conditions, the membrane potential depolarized to between -40 and -50 mV. Under these conditions, EFS caused pulse-dependent, tetrodotoxin (1 microM)-sensitive biphasic inhibitory junction potentials (IJPs) comprising a fast onset and time-to-peak phase followed by a second, slower phase that delayed repolarization. The duration of NANC IJPs ranged between 10 and 20 s. 3. Inhibition of small-conductance Ca2+-activated K+ channels with apamin (0.1 microM) selectively blocked the first fast phase of the NANC IJP, whereas inhibitors of large-conductance Ca2+-activated K+ channels (charybdotoxin and iberiotoxin) and ATP-sensitive K+ channels (glibenclamide) all had no effect on NANC IJPs. 4. Both the nitric oxide synthase inhibitor N G-nitro-L-arginine (L-NOARG; 100 microM) and the inhibitor of soluble guanylate cyclase 1-H-oxodiazol-[1,2,4]-[4,3-a] quinoxaline-1-one (ODQ; 10 microM) had no effect on the first fast phase of the NANC IJP. Each treatment, however, markedly inhibited the slow phase with the duration of the IJP reduced to between 1 and 3 s. The L-NOARG-resistant fast phase of the NANC IJP was almost abolished by the subsequent addition of apamin (0.1 microM). 5. In conclusion, the present study demonstrates unequivocal NANC nerve-mediated biphasic IJPs in the rat isolated anococcygeus. We propose that nitric oxide (NO), via activation of cGMP-dependent K+ channels, and a non-NO inhibitory factor which activates apamin-sensitive K+ channels contribute to NANC nerve-evoked IJPs in the rat anococcygeus.

Effects of suramin on electrical and mechanical activities in antrum smooth muscle of the guinea-pig stomach.

The effects of suramin on electrical and mechanical responses produced by adenosine triphosphate (ATP), acetylcholine and transmural nerve stimulation were observed in antrum smooth muscle isolated from the guinea-pig stomach. Suramin (>10(-6) M) inhibited the non-adrenergic non-cholinergic inhibitory junction potential, with no alteration of the resting membrane potential, slow wave and the ATP-induced responses (hyperpolarization and inhibition of slow waves). The amplitude, but not the frequency, of spontaneous rhythmic contraction was inhibited by suramin (>10(-5) M), with no alteration of electrical responses of the membrane. Transmural nerve stimulation elicited cholinergic excitatory and non-adrenergic non-cholinergic inhibitory responses on the spontaneous contraction, and suramin inhibited only the latter. Suramin did not alter the ATP-induced inhibition of spontaneous contraction. The contractions produced by low concentrations (<10(-7) M), but not high concentrations (10(-6) - 10(-5) M), of acetylcholine were inhibited by suramin. It is concluded that in smooth muscle of the guinea-pig antrum, suramin inhibits contractions produced spontaneously and by low concentrations of acetylcholine, with no relation to the electrical responses of the membrane. Parallel inhibition by suramin of the electrical and mechanical responses elicited by excitation of non-adrenergic non-cholinergic inhibitory nerves may not be causally related to the inhibition of ATP-receptors.

Tachykinin NK 1 and NK 2 receptors mediate non-adrenergic non-cholinergic excitatory neuromuscular transmission in the guinea-pig stomach

By using selective tachykinin NK 1 and NK 2 receptor antagonists and agonists, we studied the excitatory non-adrenergic non-cholinergic transmission to the circular muscle of the corpus of guinea-pig stomach by the sucrose-gap method. After elimination of inhibitory junction potentials by apamin (0.1 μM), l-nitroarginine (30 μM) and tetraethylammonium (10 mM), electrical field stimulation (10 Hz) in the presence of atropine (1 μM) and nifedipine (1 μM) evoked a pure excitatory junction potential and contraction. The selective tachykinin NK 2 receptor antagonist, MEN 11420, concentration-dependently inhibited the non-adrenergic non-cholinergic excitatory junction potential (EC 50=0.09 μM) and contraction (EC 50=0.04 μM) evoked by electrical field stimulation. On the other hand, the selective NK 1 receptor antagonist GR 82334 (3 μM) only slightly (by about 30%) inhibited the excitatory junction potential while leaving the contraction unaffected. The combined administration of GR 82334 (1 μM) and MEN 11420 (0.3 μM) produced an additive inhibition of the excitatory junction potential, significantly larger than that produced by each antagonist alone. In the presence of both GR 82334 (1 μM) and MEN 11420 (0.3 μM), the P 2 purinoreceptor antagonist pyridoxalphosphate-6-azophenyl-2′,4′-disulphonic acid (30 μM) remarkably inhibited the fast component of the excitatory junction potential. In the presence of atropine (1 μM), indomethacin (3 μM) and guanethidine (3 μM) either the selective NK 2 receptor agonist, [ βAla 8]neurokinin A (4–10) (0.01 μM), or the selective NK 1 receptor agonist, [Sar 9]substance P sulfone (0.3 μM), produced tetrodotoxin-(1 μM) and nifedipine-(1 μM) resistant depolarization and contraction. The [Sar 9]substance P sulfone (0.3 μM)-induced contraction, but not that induced by [ βAla 8]neurokinin A (4–10) (0.01 μM), was potentiated by apamin (0.1 μM) plus l-nitroarginine (30 μM). In the presence of atropine (1 μM), indomethacin (3 μM), guanethidine (3 μM), apamin (0.1 μM) and l-nitroarginine (30 μM), the selective tachykinin NK 2 and NK 1 receptor agonists, [ βAla 8]neurokinin A (4–10) and [Sar 9]substance P sulfone, both produced a concentration-dependent depolarization and contraction of the circular muscle. MEN 11420 inhibited the responses to [ βAla 8]neurokinin A (4–10) without affecting the responses to [Sar 9]substance P sulfone, while GR 82334 inhibited the responses to [Sar 9]substance P sulfone but not that to [ βAla 8]neurokinin A (4–10). These data provide evidence that tachykinin NK 2 receptors predominantly mediate the non-adrenergic non-cholinergic excitatory transmission to the circular muscle of the corpus of guinea-pig stomach. In addition, after blocking the non-adrenergic non-cholinergic inhibitory junction potential by apamin, l-nitroarginine and tetraethylammonium, the fast component of the non-adrenergic non-cholinergic excitatory junction potential could be mediated by adenosine triphosphate.

Tachykinin NK 1 and NK 2 receptors mediate non-adrenergic noncholinergic excitatory neuromuscular transmission in the human ileum

Tachykinin NK 1 and NK 2 receptor selective antagonists and agonists were used to study excitatory nonadrenergic non-cholinergic (NANC) transmission in circular muscle strips from human ileum by the sucrose-gap method. In the presence of atropine (1 μM), guanethidine (3 μM), indomethacin (3 μM), apamin (0.1 μM) and N ω-nitro-L-arginine (L-NOARG, 30 μM), electrical field simulation (EFS) produced a NANC inhibitory junction potential (i.j.p.) followed by NANC excitatory junction potential (e.j.p.) with superimposed action potentials and contraction of the circular muscle of human ileum. The selective tachykinin NK 1 receptor antagonist, GR 82334 (0.1–3 μM) produced a concentration-dependent inhibition of the EFS-evoked NANC e.j.p. (IC 50 = 0.21 μM) and contraction (IC 50 = 0.21 μM). The selective tachykinin NK 2 receptor antagonist, MEN 10627 (0.01–1 μM), likewise produced a concentration-dependent inhibition of the EFS-evoked NANC e.j.p. (IC 50 = 0.07 μM) and contraction (IC 50 = 0.03 μM). Either antagonist was more effective in inhibiting the mechanical than the electrical response to EFS. Neither GR 82334 nor MEN 10627 had any effect on the apamin- and L-NOARG-resistant NANC i.j.p. Activation of the NK 1 or NK 2 receptors by the selective receptor agonists, [Sar 9]substance P (SP) sulfone and [βAla 8]neurokinin A (NKA) (4–10), respectively (0.3 μM for 20 s each), produced depolarization with superimposed action potentials and contractions. GR 82334 selectively inhibited the responses to [Sar 9]]SP sulfone, without affecting the responses to [βAla 8]NKA (4–10). MEN 10627 inhibited the responses to [βAla 8]NKA (4–10), without affecting the responses to [Sar 9]SP sulfone. We conclude that both tachykinin NK 1 and NK 2 receptors co-operate in producing NANC excitation and contraction of the circular muscle in human ileum.

The possible role of ATP and PACAP as mediators of apaminsensitive NANC inhibitory junction potentials in circular muscle of guinea-pig colon.

1. In the presence of atropine (1 microM), guanethidine (3 microM), indomethacin (3 microM), nifedipine (1 microM), L-nitroarginine (L-NOARG, 100 microM), and the selective tachykinin NK1 and NK2 receptor antagonists, SR 140,333 and GR 94,800, respectively (0.1 microM each), a single pulse of electrical field stimulation (EFS) produced a monophasic non-adrenergic non-cholinergic (NANC) inhibitory junction potential (i.j.p., about 10 mV in amplitude) in the circular muscle of guinea-pig proximal colon, recorded by the modified single sucrose gap technique. 2. The P2 purinoceptor agonist, alpha, beta methylene ATP (alpha, beta mATP, 100 microM) and the pituitary adenylyl cyclase activating peptide (PACAP, 1 microM) both produced hyperpolarization (11 +/- 0.8 mV, n = 14 and 10.2 +/- 0.8 mV, n = 19, respectively) and relaxation (1.1 +/- 0.2 mV, n = 14 and 1.5 +/- 0.2 mN, n = 19, respectively) of the circular muscle. 3. Apamin (0.1 microM) nearly abolished (about 90% inhibition) the NANC i.j.p. and the alpha, beta mATP-induced hyperpolarization, markedly reduced the alpha, beta mATP-induced relaxation (73% inhibition) and the PACAP-induced hyperpolarization (65% inhibition), while the PACAP-induced relaxation was unaffected. 4. Tetraethylammonium (TEA, 10 mM) increased the EFS-evoked i.j.p. and revealed an excitatory junction potential (e.j.p.). In the presence of TEA, alpha, beta mATP induced a biphasic response: transient depolarization and contraction followed by hyperpolarization and relaxation. The hyperpolarization to PACAP was reduced by TEA (45% inhibition) but the relaxation was unaffected. 5. The combined application of apamin (0.1 microM) and TEA (10 mM) abolished the i.j.p. and single pulse EFS evoked a pure e.j.p. with latency three times longer than that of the i.j.p. In the majority of strips tested, alpha, beta mATP and PACAP elicited a biphasic response : depolarization and small contraction followed by hyperpolarization and relaxation. 6. The P2 purinoceptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) inhibited the NANC i.j.p. in concentration-dependent manner and inhibited the alpha, beta mATP-induced hyperpolarization and relaxation, without affecting the hyperpolarization and relaxation induced by PACAP. On the other hand, the P2 purinoceptor antagonist, suramin (100 microM) inhibited to a similar extent (60-80%) the NANC i.j.p. and the hyperpolarization and relaxation induced by alpha, beta mATP or PACAP. 7. PPADS and suramin reduced the NANC e.j.p. evoked by a single pulse EFS in the presence of apamin and TEA (100 microM of PPADS and 300 microM of suramin inhibited the e.j.p. by about 40%). 8. We conclude that ATP, but not PACAP, mediates the apamin-sensitive NANC i.j.p. in the circular muscle of the guinea-pig colon. After blockade of the NANC i.j.p., ATP may act as an excitatory transmitter by activating excitatory P2 purinoceptors. The subtypes of P2 purinoceptor involved in the inhibitory and excitatory responses remain to be established. The data suggest that excitatory P2 purinoceptors may be located extrajunctionally.

Characterization of the membrane conductance changes underlying the apamin-resistant NANC inhibitory junction potential in the guinea-pig proximal and distal colon

The nature of the electrically- or stretch-evoked nonadrenergic, noncholinergic (NANC) inhibitory junction potentials (UPs) in circular smooth muscle cells of the guinea-pig proximal and distal colon were investigated using standard intracellular microelectrode recording techniques. We have confirmed that the NANC UP, recorded in the presence of hyoscine (1 μM) and nifedipine (1 μM), can be divided into two components with apamin (250 nM), a blocker of the small conductance Ca 2+-activated K + channels. Both the apamin-sensitive and the apamin-resistant components of the UP were blocked by tetrodotoxin (1.6 μM) or by lowering the external Ca 2+ concentration (to 0.25 mM). The apamin-sensitive UP was also blocked by ω-conotoxin GVIA (100 nM), a blocker of ‘N-type’ Ca 2+ channels. The apamin-resistant UP and rebound post-stimulus depolarization (PSD) were reduced upon exposure to either N G- l-arginine (NOLA), an inhibitor of nitric oxide synthase (NOS), or the nitric oxide (NO) scavenger, haemoglobin. The effects of NOLA were partially reversed in the presence of excess l-arginine, a substrate for NOS, suggesting that NO, or a related NO-donor compound, is likely to be the apamin-resistant inhibitory transmitter. Blockade of either the apamin-sensitive or apamin-resistant UP was associated with membrane depolarization and a decrease in the membrane conductance in the absence of nerve stimulation. In the proximal colon, the apamin-resistant UP and PSD could both be demonstrated to arise from an increase in the membrane conductance after subtraction of a non-linear background conductance. The hyperpolarization upon repetitive NANC nerve stimulation was mimicked by the NO donor, S-nitroso- l-cysteine (2.5–25 RM), which evoked a transient apamin-sensitive, but w-conotoxin GVIA resistant, component followed by a slower apamin-resistant component. These results suggest that neurally-released NO has a number of actions in the guinea-pig colon, causing apamin-resistant hyperpolarization and depolarization, as well as directly opening apamin-sensitive K + channels.

Dual effects of trimebutine on electrical responses of gastric smooth muscles in the rat

The effects of trimebutine on the electrical properties of smooth muscle membranes were studied in the isolated rat stomach, the objective being to elucidate the dual actions of this drug on gastric motility. Transmural nerve stimulation elicited a cholinergic excitatory junction potential (e.j.p.) and a nonadrenergic noncholinergic inhibitory junction potential (i.j.p.), and trimebutine inhibited the e.j.p. more than the i.j.p., with no significant change in the acetylcholine-induced depolarization. Trimebutine reduced the interval and, at high concentrations, the amplitude of slow waves. In enzymatically dispersed single cells, the Ca 2+ current elicited by depolarization of the membrane was also inhibited by trimebutine. Thus, trimebutine increases slow wave frequency and inhibits cholinergic transmission and Ca 2+ influx. The former would enhance while the latter two would depress gastric motility.

Neuronal tachykinin NK2 receptors mediate release of non-adrenergic non-cholinergic inhibitory transmitters in the circular muscle of guinea-pig colon.

The aims of this study were: (i) verify the usefulness of the recently described non-peptide antagonist, SR 142801, for blocking tachykinin NK3 receptors in the circular muscle of the guinea-pig colon and (ii) after occlusion of NK3 receptors by SR 142801, test the hypothesis that tachykinins may activate non-adrenergic non-cholinergic inhibitory neurons via non-NK3 receptors. In sucrose gap, we found that SR 142801 (0.1 microM) time-dependently inhibited the senktide-induced atropine (1 microM)-sensitive depolarization, action potentials and contractions of circular muscle of guinea-pig colon without affecting the cholinergic excitatory junction potential and contraction produced by single pulse electrical field stimulation. Likewise, SR 142801 (0.1 microM) time-dependently inhibited the senktide-induced non-adrenergic non-cholinergic hyperpolarization and relaxation of the circular muscle, without affecting the non-adrenergic non-cholinergic inhibitory junction potentials and relaxation produced by single pulse electrical field stimulation. Therefore, SR 142801 is a suitable tool to occlude neuronal NK3 receptors in guinea-pig colon. In the presence of SR 142801 (0.1 microM), atropine (1 microM), guanethidine (3 microM), indomethacin (3 microM) and nifedipine (1 microM) superfusion with neurokinin A (0.3 microM) produced depolarization on which a series of inhibitory junction potentials were superimposed. The incidence, number and amplitude of the inhibitory junction potentials evoked by neurokinin A was partly reduced by pretreatment with either apamin (0.1 microM) or L-nitroarginine (30 microM) and was totally blocked by pretreatment with apamin plus L-nitroarginine or by tetrodotoxin (1 microM). None of these treatments affected the depolarization and contraction produced by neurokinin A. The NK1 receptor selective antagonist, GR 82,334 (3 microM), did not affect the responses to neurokinin A, which were abolished by the NK2 receptor-selective antagonist GR 94,800 (0.1 microM). Substance P (0.3 microM) produced a large depolarization of the membrane but was poorly effective in producing superimposed inhibitory junction potentials. The NK1 receptor-selective agonist [Sar9]substance P sulfone (0.3 microM) produced large depolarization without inducing superimposed inhibitory junction potentials, while the NK2 receptor-selective synthetic agonist [beta-Ala8]neurokinin A(4-10) (0.3 microM) produced depolarization and superimposed inhibitory junction potentials. We conclude that neurokinin A, in addition to direct excitation and contraction of circular muscle activates, via neuronal NK2 receptors, inhibitory non-adrenergic non-cholinergic motorneurons. Thus, neuronal NK2 receptors should be considered as targets for endogenous tachykinins in enteric circuitries leading to descending relaxation in guinea-pig colon.

Excitation and inhibition by cisapride of spontaneous and evoked electrical responses in smooth muscles of guinea-pig stomach

In isolated circular smooth muscles of the guinea-pig stomach, cisapride depolarized the membrane, increased the amplitude and interval of slow waves, and enhanced the cholinergic excitatory junction potential, with no change in the nonadrenergic noncholinergic inhibitory junction potentials. Methysergide mimicked the excitatory actions of cisapride on junction potentials. Results showed that cisapride has dual actions on gastric muscles; excitation by facilitating the release of ACh possibly via 5-HT receptor blockade and also by depolarizing the smooth muscle membrane, and inhibition by reducing the frequency of slow waves, possibly by acting directly on the pacemakers.

Evidence that tachykinin NK1 and NK2 receptors mediate non-adrenergic non-cholinergic excitation and contraction in the circular muscle of guinea-pig duodenum.

1. In the presence of atropine (1 microM), guanethidine (3 microM), indomethacin (3 microM), apamin (0.1 microM) and L-nitroarginine (L-NOARG, 30 microM), electrical field simulation (EFS) produced a nonadrenergic, noncholinergic (NANC) excitatory junctional potential (e.j.p.), action potentials and contraction of the circular muscle of the guinea-pig proximal duodenum, recorded by the single sucrose gap technique. 2. The selective tachykinin (TK) NK1 receptor antagonist, GR 82,334 (30 nM-3 microM) produced a concentration-dependent inhibition of the EFS-evoked NANC e.j.p. and contraction. Similarly, the selective NK2 receptor antagonists, MEN 10,627 (30 nM-3 microM) and GR 94,800 (100 nM-10 microM), both produced a concentration-dependent inhibition of the EFS-evoked NANC e.j.p. and contraction. GR 82,334 inhibited the electrical and mechanical NANC responses to EFS in an almost parallel manner, while MEN 10,627 and GR 94,800 were more effective in inhibiting the mechanical than the electrical response to EFS. 3. Activation of the NK1 or NK2 receptor by the selective agonists, [Sar9]substance P (SP) sulphone and [beta Ala8]neurokinin A (NKA) (4-10), respectively (0.3 microM each), produced depolarization, action potentials and contractions. GR 82,334 selectively inhibited the responses to [Sar9]SP sulphone, without affecting the responses to [beta Ala8]NKA (4-10). MEN 10,627 and GR 94,800 inhibited or abolished the responses to [beta Ala8]NKA (4-10), without affecting the responses to [Sar9]SP sulphone. 4. Nifedipine (1 microM) abolished the action potentials and contraction produced either by EFS or by the TK receptor agonists [Sar9]SP sulphone or [beta Ala8]NKA (4-10). 5. In the presence of nifedipine, the NANC e.j.p. produced by EFS was biphasic: in the majority of strips tested (21 out of 29) an early fast phase of depolarization was followed by a second slow component. The combined administration of GR 82,334 and GR 94,800 (3 microM each) reduced both components, the slow phase being inhibited to a greater extent than the fast phase. 6. The P2 purinoreceptor antagonist, suramin (100 microM) reduced the fast phase of the e.j.p. produced by EFS in the presence of nifedipine, without affecting the slow phase. The combined administration of suramin, GR 82,334 and GR 94,800 produced a nearly complete blockade of the e.j.p. produced by EFS in the presence of nifedipine. 7. When tested in the absence of apamin and L-NOARG, EFS induced a NANC inhibitory junction potential (i.j.p.) followed by an e.j.p., and the selective P2Y receptor agonist, adenosine-5'-O-(2-thiodiphosphate) (ADP beta S, 10 microM), produced membrane hyperpolarization. After addition of apamin and L-NOARG, the ij.p. was blocked, and EFS produced a pure NANC e.j.p.; ADPPS produced depolarization, action potentials and contraction.8. Suramin (100 microM) blocked the depolarization, action potentials and contractions produced by ADP beta S in the presence of apamin and L-NOARG, without affecting the responses produced by the NK1receptor agonist, [Sar9}SP sulphone.9. We conclude that NK1 and NK2 receptors cooperate in producing NANC excitation and contraction of the circular muscle in the guinea-pig proximal duodenum. Activation of either TK receptor produces membrane depolarization and both receptors contribute to generate action potentials which are essential for producing muscle contraction, via nifedipine-sensitive calcium channels. It appears that endogenous ATP chiefly acts as an inhibitory transmitter but, after blockade of NANC inhibitory mechanism(s),ATP may act as a fast signalling excitatory transmitter.

Noradrenergic, noncholinergic inhibitory junction potentials in rat proximal colon: role of nitric oxide

Using a single sucrose gap apparatus, experiments were performed to determine the involvement of nitric oxide (NO) in the generation of nonadrenergic, noncholinergic (NANC) inhibitory junction potentials in circular muscle of rat proximal colon. Inhibitors of NO synthase, N omega-nitro-L-arginine and its methyl ester, reduced the amplitude of the electrically evoked inhibitory junction potentials, without affecting membrane resting potential. Such an effect was stereospecific and it was prevented by L-arginine but not by D-arginine. Sodium nitroprusside induced a tetrodotoxin-resistant hyperpolarization, which was not affected by NO synthase inhibitors. Apamin reduced sodium nitroprusside induced hyperpolarization, as well as NANC inhibitory junction potentials, and alpha-chymotrypsin decreased the amplitude of electrical field stimulation evoked responses. Residual responses after NO synthase inhibitors or after alpha-chymotrypsin were further reduced by pretreatment with alpha-chymotrypsin or NO synthase inhibitors, respectively. These results suggest that, in rat colonic circular muscle, NO plays an important role in NANC inhibitory junction potential generation. However, another mechanism, peptidergic in nature, is also involved.

Suramin selectively inhibits the non-adrenergic non-cholinergic inhibitory junction potential in the guinea-pig stomach

In smooth muscle cells of the guinea-pig stomach fundus, transmural nerve stimulation evoked a cholinergic excitatory junction potential (e.j.p.) and, in the presence of atropine, a non-adrenergic non-cholinergic (NANC) inhibitory junction potential (i.j.p.). Suramin (> 10 −5 M), a putative inhibitor of the P 2 purinoceptor, enhanced the e.j.p. amplitude and inhibited the i.j.p., with no significant effect on the membrane potential. Thus, a possible involvement of ATP in the generation of the NANC i.j.p. has to be considered.

Nitric oxide and nitrosocysteine mimic nonadrenergic, noncholinergic hyperpolarization in canine proximal colon.

Previous evidence suggests that nonadrenergic, noncholinergic (NANC) inhibitory neurotransmission in visceral muscles may be mediated by nitric oxide (NO). We have demonstrated that NO and the NO carrier S-nitrosocysteine can mimic the hyperpolarization in colonic muscle caused by nerve stimulation. The finding that S-nitrosocysteine breaks down fast enough to cause inhibitory junction potential (IJP)-like hyperpolarizations suggests that NO could be stored as a nitrosothiol in secretory vesicles in nerve terminals. Oxyhemoglobin blocked hyperpolarization responses to NO and S-nitrosocysteine and NANC IJPs. These findings suggest that NO is a biologically active transmitter substance in NANC inhibitory neurotransmission. NO enhanced the open probability of Ca(2+)-activated K+ channels in isolated colonic muscle cells. These channels may mediate the hyperpolarization response to NANC neurotransmission in colonic muscles.

Modulation of smooth muscle activity by excitatory and inhibitory nerves in the guinea-pig stomach

1. 1. In smooth muscle of the guinea-pig stomach, intramural nerve stimulation evoked cholinergic excitatory junction potential in the fundus and non-adrenergic non-cholinergic inhibitory junction potential in the antrum, yet cholinergic contractions in both regions. 2. 2. This dissociation between electrical and mechanical responses was mainly due to different sensitivity of the membrane for depolarization to acetylcholine.