From biological gastroenterology to fundamental neurosciences: how studies in gastric emptying have led to the discovery of a new mechanism of neuronal functioning.

Gastric emptying undergoes complex regulation by the nervous system, which organizes in particular the inhibition of duodenum motility after a rise in intra-gastric pressure: the gastro-duodenal inhibitory reflex. It was first shown in mammals that this reflex could be organized by a sympathetic ganglion, the coeliac plexus. The excitation of gastric mechanosensitive fibres leads in this ganglion to the release of a neurotransmitter, which in turn activates ganglionic neurones leading to inhibition of the duodenum contractions. It rapidly became apparent that this reflex presented striking properties since it was organized in the absence of action potentials along the nerve fibres. Then it was shown that the neurotransmitter released in the coeliac plexus was gaseous: nitric oxide (NO). The nature of the mechanism conducting, without action potentials, the excitation along the nerve fibres was recently determined. This mechanism necessitates the integrity of particular areas of the neuronal membrane (the lipid rafts) and the activation in cascade of the following second messenger sequence: ceramide, calcium, NO and guanosine cyclic monophosphate (c-GMP). These results show how studies in biological gastroenterology have led to the rethinking of one of the central dogmas in neuroscience according to which excitation is only conducted along the nerves by the action potential.

Modulation by C2 ceramide of the nicotinic transmission within the coeliac ganglion in the rabbit.

We have investigated the modulation by ceramide of the nicotinic activation of the prevertebral sympathetic neurons. Our study was performed in vitro in rabbit isolated coeliac ganglion, using intracellular recording techniques. We have used C(2) ceramide, a permeant analog of ceramide. The effects of C(2) ceramide were first assessed when nicotinic activation was elicited without modulatory mechanisms (fast excitatory postsynaptic potentials triggered by stimulation of the thoracic splanchnic nerves with a single pulse). In all the neurons tested, C(2) ceramide triggered an increase in the amplitude of the fast excitatory postsynaptic potentials demonstrating a direct facilitatory effect on the nicotinic activation. We then investigated the effects of C(2) ceramide on modulatory mechanisms of this activation. These mechanisms occur when a train of pulses of supramaximum intensity is applied on the splanchnic nerves. During the train, a gradual depression of fast nicotinic activation occurred: the pulses failed to systematically elicit action potentials. We have previously demonstrated that this regulatory phenomenon is partly modulated by nitric oxide which exerts a dual effect: facilitation or inhibition of the nicotinic activation. In all the neurons tested, C(2) ceramide decreased the number of action potentials fired during a train of pulses, demonstrating an indirect inhibitory effect on the nicotinic activation. The use of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (nitric oxide scavenger) suppressed the inhibitory effect of C(2) ceramide, demonstrating that this effect is mediated through the nitric oxide pathway. C(2) dihydro-ceramide, an inactive analog of ceramide, was without effect on the nicotinic activation of the ganglionic neurons. These results demonstrate that ceramide exerts a complex modulation of the nicotinic activation of the prevertebral neurons: direct facilitation and indirect inhibition involving the nitric oxide pathway. In fact, C(2) ceramide plays a key gating role in the dual effect of the nitric oxide pathway by activating the inhibitory effect. The existence of this gating mechanism involving ceramide and nitric oxide opens new perspectives in terms of our understanding of the modulation of synaptic transmission within the prevertebral ganglia. Our study demonstrates that sphingolipids are involved in complex modulations of the synaptic activation within the prevertebral ganglia, and thus contribute to their integrative properties.

Effects of testosterone on the electrical properties and nicotinic transmission of the major pelvic and coeliac ganglion neurones.

The effects of testosterone on the electrical properties and nicotinic activation of prevertebral ganglion neurones were investigated in vitro on the male rat major pelvic ganglion and rabbit coeliac ganglion. The electrical activity of the neurones was recorded using intracellular recording techniques. Nicotinic activation was triggered for neurones of the major pelvic ganglion by stimulating the hypogastric, pelvic and cavernous nerves and for coeliac neurones by stimulating the splanchnic nerves. Testosterone modified the resting membrane potential of neurones in the major pelvic ganglion by triggering a slow depolarization, and was without significant effect on the resting membrane potential of coeliac ganglion neurones. In neurones of the major pelvic and coeliac ganglia, testosterone had no significant effect on the firing pattern, on the characteristics of the action potential (firing threshold, duration, overshoot) and on the after-hyperpolarization (amplitude and duration). Testosterone affected, in opposite ways, the nicotinic activation of neurones of the two prevertebral ganglia. In the major pelvic ganglion, testosterone triggered an increase in the amplitude of excitatory postsynaptic potentials induced by stimulation of the hypogastric, pelvic and cavernous nerves with a single pulse, revealing a facilitation of nicotinic activation. On coeliac ganglion neurones, testosterone elicited a decrease in the amplitude of excitatory postsynaptic potentials induced by stimulation of the splanchnic nerves, indicating an inhibition of nicotinic activation. Our study shows that testosterone acts differently on neurones of prevertebral ganglia involved in the nervous control of different functions, its facilitatory action being exerted on neurones of the major pelvic ganglion which is particularly involved in the control of the urogenital tract. Our study reinforces the concept, derived from neuroanatomical and pharmacological studies, of the major pelvic ganglion as a major peripheral target for testosterone.

Nitric oxide released by gastric mechanoreceptors modulates nicotinic activation of coeliac plexus neurons in the rabbit.

The effects on the nicotinic activation of the coeliac plexus neurons of nitric oxide (NO) released within the coeliac plexus by gastric mechanoreceptors, in particular during gastroduodenal inhibitory reflex, were assessed. This study was performed in the rabbit on an in vitro preparation of the coeliac plexus connected to the stomach and the duodenum. The electrical activity of ganglionic neurons was recorded with intracellular recording techniques. Water-filled balloons were used for gastric distensions and recording of duodenal motility. When a 10-s train of pulses (20-40Hz) of supramaximal intensity was applied to the splanchnic nerves, gradual depression of nicotinic activation occurred. Gastric distension (50 mL, 7.5 min) modulated this depression phenomenon by inhibiting or facilitating the nicotinic activation. In the neurons impaled during the recording of duodenal motility, gastric distension triggered an inhibition of nicotinic activation concomitantly with a gastroduodenal inhibitory reflex organized by the coeliac plexus. If the gastric distensions were performed while the coeliac plexus was superfused by a NO scavenger, the nicotinic activation was unaffected and the gastroduodenal inhibitory reflex was abolished. Moreover, when the coeliac plexus was superfused with an inhibitor of nitric oxide synthase, gastric distensions were without effect on the nicotinic activation. These results demonstrate that NO released within the coeliac plexus by gastric mechanoreceptors, in particular during the gastroduodenal inhibitory reflex, modulates the central nicotinic activation of coeliac plexus neurons, so NO released within a prevertebral ganglion by gastric afferent fibres, in particular during the organization by this ganglion of a reflex regulating the gastrointestinal tract motility, also exerts a gating of the central inputs to the ganglionic neurons.

Muscarinic receptor activation is a prerequisite for the endogenous release of nitric oxide modulating nicotinic transmission within the coeliac ganglion in the rabbit.

The aim of the present study was to investigate whether the activation of muscarinic receptors is a preliminary step to the endogenous release of nitric oxide modulating nicotinic transmission within the prevertebral ganglia. This work has been performed in vitro in isolated rabbit coeliac ganglion. The electrical activity of the ganglionic neurons was recorded using intracellular recording techniques. When a train of pulses of supramaximal intensity was applied to the splanchnic nerves, gradual depression of fast nicotinic transmission occurred: the pulses do not systematically elicit action potentials, but very often elicit excitatory postsynaptic potentials only. The use of pharmacological agents that interfere with the nitric oxide pathway such as L-arginine (precursor of nitric oxide) or 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (nitric oxide scavenger) demonstrated that nitric oxide modulates this depression phenomenon by facilitating or inhibiting the nicotinic transmission of the ganglionic neurons. A nitric oxide donor (diethylamine/nitric oxide complex) induced an inhibition of the nicotinic synaptic transmission. In the presence of the muscarinic receptors antagonist atropine, L-arginine and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide failed to modify the nicotinic transmission of the ganglionic neurons but diethylamine/nitric oxide complex was still able to inhibit it. These results demonstrate that in the coeliac ganglion, the activation of muscarinic cholinergic receptors is a prerequisite for the activation of neuronal nitric oxide synthase in preganglionic fibres. The nitric oxide released then exerts a facilitation or an inhibition of the nicotinic transmission of the ganglionic neurons. Atropine triggered a facilitation of the nicotinic transmission when superfused alone and an inhibition when superfused in the presence of 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide. These results confirm that muscarinic receptors activate the nitric oxide pathway modulating the nicotinic transmission of the prevertebral neurons. Our results also demonstrate that when the nitric oxide pathway is blocked, activation of muscarinic receptors leads to facilitation of the nicotinic transmission. Our study brings new insights concerning the modulation by nitric oxide and by muscarinic receptors of the synaptic transmission within the prevertebral ganglia.

Release of nitric oxide within the coeliac plexus is involved in the organization of a gastroduodenal inhibitory reflex in the rabbit.

1. The coeliac plexus can organize a gastroduodenal inhibitory reflex without action potentials. The involvement of the nitric oxide-cGMP pathway in this reflex was investigated in the rabbit on an in vitro preparation of the coeliac plexus connected to the stomach and duodenum. Intraluminal duodenal pressures were measured with water-filled balloons. Gastric distension inhibited duodenal motility, thus characterizing a gastroduodenal inhibitory reflex organized by the coeliac plexus. 2. L-Arginine, superfused at the coeliac plexus level, enhanced this reflex, whereas Nomega-nitro-L-arginine (L-NOARG) or 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (carboxy PTIO) reduced or abolished it. Moreover, diethylamine/nitric oxide complex superfused at the coeliac plexus level inhibited duodenal motility in the absence of gastric distension. 3. The effects of nitric oxide were mediated through the activation of guanylyl cyclase, as 1H-[1,2,4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ) reduced or abolished the gastroduodenal inhibitory reflex, whereas zaprinast enhanced it. Moreover, 8-bromo-cGMP and cGMP, superfused at the coeliac plexus level, inhibited duodenal motility in the absence of gastric distension. 4. On the other hand, when perfused at the visceral level, L-NOARG, propranolol plus phentolamine, and guanethidine did not affect the reflex. Thus, neither nitric oxide nor noradrenaline could be the transmitters released at the muscular level to induce this reflex. 5. Our study demonstrates that the gastroduodenal inhibitory reflex, which is organized by the coeliac plexus without action potentials, is induced by the release within the plexus of nitric oxide acting on the cGMP pathway. These results provide new insights into the control of digestive motility by the prevertebral ganglia.

Integrative properties of the major pelvic ganglion in the rat.

The integrative properties of the major pelvic ganglion were investigated in the male rat on an in vitro preparation consisting of the ganglion connected to the hypogastric, pelvic and cavernous nerves. The electrical activity of ganglionic neurones was recorded using intracellular recording techniques. The neurones never displayed any spontaneous activity and were found to be only of the phasic type. Fast synaptic activation could be evoked in the same neurone by stimulating the hypogastric, pelvic and cavernous nerves with a single pulse. This activation was not affected by hexamethonium plus D-tubocurarine but was abolished by mecamylamine. During nerve stimulation with a train of pulses, a gradual depression of the fast synaptic responses occurred. This phenomenon increased with the frequency of stimulation. Our results show that the neurones of the major pelvic ganglion can integrate central inputs from both the sympathetic and parasympathetic systems as well as peripheral inputs. This activation is modulated by a rate limiting mechanism. Thus the major pelvic ganglion should not be considered as a simple relay but as a true integrative nervous centre which opens new perspectives concerning its role in the nervous control of the urogenital and gastrointestinal tracts.

Nerve-induced release of nitric oxide exerts dual effects on nicotinic transmission within the coeliac ganglion in the rabbit.

The involvement of nitric oxide in the modulation of nicotinic activation was investigated in vitro in isolated rabbit coeliac ganglion. The electrical activity of the ganglionic neurons was recorded using intracellular recording techniques. When a train of pulses of supramaximum intensity was applied to the splanchnic nerves, gradual depression of fast nicotinic activation occurred: the pulses do not systematically elicit action potentials, but very often elicit excitatory postsynaptic potentials only. This phenomenon appeared between 15 and 20 Hz and increased with the frequency of stimulation. It was not related to any change in the membrane potential of the ganglionic neurons. For a given frequency, the depression appeared progressively and it was particularly strong at the end of the train. The use of pharmacological agents that interfere with the nitric oxide pathway, such as L-arginine (precursor of nitric oxide), D-arginine (non-precursor of nitric oxide) N(omega_-nitro-L-arginine and N(omega)-nitro-L-arginine methyl ester (inhibitors of nitric oxide synthase), and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (nitric oxide scavenger), demonstrated that nitric oxide modulated this depression phenomenon by exerting a dual effect on the nicotinic activation, i.e. facilitation or inhibition. Agents interfering with the guanosine 3',5'-cyclic monophosphate pathway, such as oxadiazolo[4,3-a] quinoxalin-1-one (selective inhibitor of the nitric oxide-activated soluble guanylate cyclase) and zaprinast (selective inhibitor of the phosphodiesterases involved in the guanosine 3',5'-cyclic monophosphate pathway) demonstrated that only the facilitatory effect of nitric oxide on the nicotinic activation was mediated through the guanosine 3',5'-cyclic monophosphate pathway. The mechanism sustaining the inhibitory effect remains to be determined. By modulating the nicotinic activation, nitric oxide plays a role in the integrative properties of the prevertebral ganglia. This opens new perspectives with regard to the control of visceral functions by the prevertebral level of regulation.

The mammalian sympathetic prevertebral ganglia: integrative properties and role in the nervous control of digestive tract motility.

The prevertebral ganglia which are a constitutive part of the sympathetic system have long been considered as a simple relay on this efferent pathway. In fact, these ganglia must be considered as true peripheral nervous centres. They possess various integrative properties, such as projections of central and peripheral inputs onto the ganglionic neurones, gating of these projections and pacemaker activity of the ganglionic neurones. These properties explain the ability of these ganglia to participate in the regulation of various visceral functions, including digestive tract motility.

Muscarinic activation of a novel voltage-sensitive inward current in rabbit prevertebral sympathetic neurons.

The muscarinic activation of rabbit prevertebral sympathetic neurons was studied in non-dissociated coeliac and superior mesenteric ganglia using whole-cell patch-clamp techniques. In the presence of nicotinic blockers, carbachol, muscarine and oxotremorine-M (1-50 microM) induced tonic firing by activating a persistent inward current. These effects were abolished by atropine. They persisted when the M-current was blocked with Ba2+ (1 mM) and intracellular Cs+. The muscarinic inward current was found to be time- and voltage-dependent. It peaked at -60 mV, decreased at large hyperpolarizations and was tonically activated between -110 and -20 mV, which gave steady-state I-V curves an N-shape between -96 and -54 mV. The negative slope accounted for the large hyperpolarizing responses generated by current pulses in carbachol-treated cells. The muscarinic current was abolished when Na+ was replaced by choline, Tris+, sucrose, N-methyl-D-glucamine and Cs+ but not Li+. It was resistant to tetrodotoxin (3 microM), amiloride (3 microM), benzamil (10 microM) and tetraethylammonium (5-20 mM). No involvement of K+ and Cl- could be detected. We therefore styled it INa,M, in reference to its ionic selectivity and its coupling to muscarinic receptors. Low Ca(2+)-Mg2+ salines enhanced the Na,M-current. The current was blocked by Cd2+, Co2+, La3+ (1 mM) and Ba2+ (5 mM) but insensitive to methoxyverapamil hydrochloride, nicardipine, nifedipine and omega-conotoxin MVII A (2-20 microM). These effects were ascribed to the binding of di- and trivalent ions to the Na,M-channels. Spike bursts transiently blocked INa,M. With high intracellular ethylene glycol bis(b-aminoethyl ether)-N,N'-tetraacetic acid or 1,2-bis (2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (20-50 mM), this effect was reduced, whereas INa,M persisted in long-term recordings and its amplitude increased twofold, indicating that intracellular calcium negatively regulated the Na,M-channels. We conclude that we have described a novel muscarinic receptor-coupled channel which appears to play a major part in regulating the firing behaviour of sympathetic neurons.

Synaptically activated low-threshold muscarinic inward current sustains tonic firing in rabbit prevertebral sympathetic neurons.

Whole-cell patch-clamp experiments were performed on non-dissociated rabbit coeliac sympathetic neurons in the presence of nicotinic blockers. Coeliac neurons were classified as either silent or spontaneously active (pacemaker) cells. Under voltage-clamp conditions, pacemaker cells exhibited a steady-state N-shaped current-voltage relationship due to the presence of a persistent voltage-dependent inward current in the potential range of -100 to approximately -20 mV. This inward current sustained the regular firing activity of pacemaker cells and was absent from quiescent neurons. It disappeared in the presence of tetrodotoxin and in low Ca(2+)-high Mg2+ external solutions and was enhanced by eserine. Splanchnic nerve stimulation induced slow regenerative depolarizations and firing discharges in silent neurons by activating a low-threshold voltage-sensitive inward current. The synaptic current had a U-shaped voltage-dependence from -96 to approximately -20 mV and exhibited the dynamic properties of the muscarinic voltage-dependent inward current INa,M. It gave the current-voltage relationship an N shape similar to that observed in spontaneously active cells. The muscarinic antagonists atropine and pirenzepine abolished the inward current present in pacemaker cells and that induced by nerve stimulation in silent neurons. These data provide evidence that both spontaneous firing activity and nerve-evoked depolarizing responses in coeliac neurons are sustained by the activation of the muscarinic Na,M current. The tonic activation of INa,M in spontaneously firing cells results from a sustained Ca(2+)-dependent tetrodotoxin-sensitive release of acetylcholine. This study provides evidence that the role of the muscarinic receptors is not purely a neuromodulatory one, but that these receptors are directly involved in ganglionic neurotransmission.

Immunochemical study of galanin in the cat digestive tract and autonomic ganglia.

The presence of galanin was examined in the cat gut and related autonomic nervous structures using radioimmunoassay (RIA) and high performance liquid chromatography (HPLC). In the gut wall, the concentration of galanin-like immunoreactivity (GAL-Lt) was assayed separately in the muscular layers with the nervous plexuses and in the mucosa and ranged from 0.35 to 0.55 pmol/g wet tissue. In the autonomic nervous structures, GAL-L1 concentrations ranged from 0.22 (thoracic spinal ganglia) to 0.81 (inferior mesenteric ganglion) pmol/g wet tissue. The presence of galanin was checked by HPLC in the antrum, intestine, and colon. HPLC of extractable material revealed a major peak coeluting with the synthetic porcine peptide and minor earlier peaks representing likely different molecular forms of galanin. Our study strengthens the notion that galanin acts in nervous control of the cat gut functions.

New insights into the organization of a gastroduodenal inhibitory reflex by the coeliac plexus.

The mechanisms involved at the prevertebral ganglionic level in a gastroduodenal inhibitory reflex were investigated in the rabbit on an in vitro preparation of the coeliac plexus connected to the stomach and duodenum. Intraluminal gastric and duodenal pressures were measured using water-filled balloons. Gastric distension inhibited duodenal motility via a nerve reflex which was abolished by section of the nerves connecting the coeliac plexus to the viscera. Superfusion of the coeliac plexus with a low Ca(2+)-high Mg2+ solution abolished the gastroduodenal inhibitory reflex, indicating a synaptic link at the ganglion level. The reflex was unaffected by superfusion of the coeliac plexus with hexamethonium and tubocurarine, ruling out a nicotinic mechanism. The reflex persisted when the coeliac plexus was superfused with tetrodotoxin or when the nerves connecting the coeliac plexus to the viscera were superfused with a Na(+)-free solution; these results indicate that the reflex does not involve sodium-dependent action potentials. Moreover, superfusion of the nerves connecting the coeliac plexus to the viscera with a calcium blocker or with a Ca(2+)-free solution also failed to abolish the reflex, suggesting that calcium-dependent action potentials are not involved. Our study demonstrates that a gastrointestinal inhibitory reflex via the coeliac ganglion is not based on fast synaptic inputs or action potentials. These results provide new insights concerning the physiology of the sympathetic prevertebral ganglia.

Satellite glial cells in situ within mammalian prevertebral ganglia express K+ channels active at rest potential.

Patch-clamp experiments were performed on satellite glial cells wrapped around sympathetic neurons in the rabbit coeliac ganglion. With the cleaning method used, the glial cells could be kept in place and were directly accessible to the patch-clamp pipettes. Whole-cell recordings showed that glial cells had almost ohmic properties. Their resting potential (-79.1 +/- 1.2 mV) was found to be very nearly the same as the K+ reversal potential and approximately 20 mV more negative than that of the neurons they encapsulated. Unitary currents from ionic channels present in the glial membrane were recorded in the cell-attached configuration with pipettes filled with various amounts of K+, Na+ and gluconate. Only K(+)-selective channels with slight inwardly rectifying properties (in the presence of 150 mM [K+]o) were detected. These channels were active (Po = 0.7-0.8) at the cell resting potential. The channel conductance, but not its opening probability, was dependent on the [K+] in the pipette. Cl(-)-selective channels (outwardly rectifying and large conductance channels) were detected in excised patches. The properties of the K+ channels (increased inward current with [K+] and detectable outward current at low [K+]) are well suited for siphoning the K+ released by active neurons.

Inhibition of peripheral fast synaptic inputs to celiac ganglion neurons by splanchnic preganglionic fibers in the cat.

Fast nicotinic transmission was studied in vitro in isolated cat celiac ganglion neurons using intracellular recording techniques. Fast synaptic activation was evoked by stimulation (0.1 Hz) of the anterior peripheral rami. A long-lasting inhibition of this response was triggered by repetitive splanchnic stimulation (30-50 Hz) for 10 s. Evidence is given that this inhibition occurs at presynaptic level. Our results indicate that central inputs modulate transmission of fast synaptic peripheral inputs to prevertebral ganglionic neurons. This would be another integrative mechanism in the prevertebral ganglia.

Electrical and integrative properties of rabbit sympathetic neurones re-evaluated by patch clamping non-dissociated cells.

1. Voltage recordings were performed on non-dissociated sympathetic neurones from rabbit coeliac ganglia using the whole-cell configuration of the patch clamp technique. 2. Cells were classified depending on their firing pattern as silent cells (63%) producing either phasic (24%) or tonic (76%) spike discharge in response to depolarizing currents, and pacemaker cells (37%). 3. All the cells produced large overshooting spikes and prolonged postspike after-hyperpolarization. The peak-to-peak spike amplitude was 113.8 +/- 1 mV. Spikes were shortened and the after-hyperpolarization was suppressed when calcium channel blockers (Cd2+ and La3+) were added. 4. Silent cells have a resting potential of -58.8 +/- 1.5 mV. At potentials ranging from -50 to -90 mV, the input impedance was 490 +/- 27 M omega at 22-24 degrees C and 426 +/- 47 M omega at 35-36 degrees C. The time constant at voltages corresponding to the high input impedance region was 126 +/- 7 ms at 22-24 degrees C and 86 +/- 7 ms at 35-36 degrees C. 5. The firing frequency of the pacemaker cells was 3.2 +/- 0.5 Hz at 35-36 degrees C in the presence of nicotinic blockers. Evidence is given that the firing did not result from cell injury but was induced by an intrinsic pacemaker mechanism. Input impedance of pacemaker neurones was 580 +/- 47 M omega at 22-24 degrees C and 473 +/- 56 M omega at 35-36 degrees C. 6. Most of the pacemaker cells (63%) were motoneurones, since they were antidromically fired by stimulating post-ganglionic nerves. In addition, they received synaptic inputs from both preganglionic fibres (splanchnic nerves) and the periphery (postganglionic nerves). Long-lasting depolarizations were induced in either silent or pacemaker cells by single shocks applied to pre- and postganglionic nerves. 7. Slowly rising voltage ramps revealed the presence of an N-shaped current-voltage relationship in voltage clamped pacemaker cells. The negative slope was located in a subthreshold voltage range, between -83.4 +/- 1.4 and -59.0 +/- 1.8 mV. It was induced by the activation of a low threshold persistent inward current. Although it was tiny (22 +/- 3 pA at its peak level) this current brought the null-current voltage up to -41.0 +/- 1.4 mV, which resulted in continuous firing. 8. Due to the instability introduced by the N-shaped I-V relationship, pacemaker cells can display bistable behaviour characterized by hyperpolarizing responses.(ABSTRACT TRUNCATED AT 400 WORDS)

Single-channel and whole-cell recordings from non-dissociated sympathetic neurones in rabbit coeliac ganglia.

A procedure is described for performing patch-clamp recordings on mammalian sympathetic neurones within intact ganglia. The plasma membrane of superficial neurones was cleaned by blowing (1.5-3 h) a gentle stream of Ringer saline onto ganglia, the connective sheath of which was previously softened by a short protease treatment. This procedure preserved the intraganglionic connectivity so that the neurones could be activated either synaptically or antidromically by stimulating the appropriate nerves. Depending on the duration of the mechanical cleaning step, recordings were performed on either the neurones or the satellite glial cells covering the neuronal cell bodies. The applicability of the various configurations of the patch-clamp technique to studying sympathetic neurones is illustrated by recordings of whole-cell voltage, whole-cell currents and single-channel currents in cell-attached and excised patches. With these techniques, the resolution of the membrane current recordings is higher than with conventional microelectrodes. The results obtained show that mammalian sympathetic neurones have a very high input resistance (0.5 G omega), are electronically compact and may display pacemaker activity. These techniques provide a useful tool for studying the synaptic transmission and neuromodulation mechanisms operating within the sympathetic ganglia.

[Role of substance P in the nervous system control of digestive motility]. / Rôle de la substance P dans le contrôle nerveux de la motricité digestive.

Substance P is a 11 amino-acids peptide which belongs to the tachykinins, a family of peptide which induces a rapid contraction of the smooth muscle of the digestive tract. The occurrence of substance P has been demonstrated by immunohistochemical and radioimmunological techniques in most parts of the central and peripheral nervous system. Substance P exerts on the smooth muscle of all the areas of the digestive tract a strong excitatory effect which is either direct or relayed by the cholinergic intramural neurones. Numerous electrophysiological, pharmacological and immunohistochemical data lead to the conclusion that substance P is released by intrinsic neurones of the digestive tract or by extrinsic nerves (vagus and splanchnic nerves, etc...). This release is enhanced by acetylcholine, cholecystokinin, serotonin and neurotensin, it is reduced by opioid peptides and noradrenaline. Substance P participates in the intestinal peristaltic reflex by the activation of the smooth muscle cells of the intestine, either directly or through the activation of the cholinergic intrinsic neurones. Substance P is also involved in the genesis of a non-cholinergic ascending excitatory activity likely occurring during vomiting. Lastly, substance P participates in the reflex contraction of the lower oesophageal sphincter following acidification of the distal part of the oesophagus.