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J J Puizillout

Publications and source records attributed to J J Puizillout.

At least 19 recordsLinked to original sources
PubMed1998

Substance P post-synaptically potentiates glutamate-induced currents in dorsal vagal neurons.

We examined the post-synaptic actions of glutamate, N-methyl-D-aspartate (NMDA) and substance P on dorsal vagal neurons, using the patch-clamp technique on brainstem slices of young rats. The vagal neurons were identified electrically and histologically. All vagal neurons responded to glutamate and NDMA and about 30% to substance P, with dose-dependent inward currents. The glutamate-induced currents were blocked partially by either CPP (3((R)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid) or CNQX (6-cyano-7-nitro-quinoxaline-2,3-dione), indicating that these currents resulted from the activation of at least two types of glutamate receptors: NMDA receptors and AMPA/kainate receptors. The NK1 receptor-selective antagonist, RP67580, blocked substance P-induced currents, suggesting that NK1 receptors do coexist with NMDA receptors and AMPA/Kainate receptors. Substance P potentiated the effects of glutamate. This potentiation lasted 10-20 min and was blocked by CPP and by RP67580, but not by CNQX, demonstrating that the increase in glutamate-induced currents resulted from the interaction between NK1 receptors and NMDA channels. These results provided the first evidence that the receptors for substance P and glutamate coexist on dorsal vagal neurons and interact with each other to modulate visceral efferent functions.

Animals↗
PubMed1996

Effects of tachykinins on identified dorsal vagal neurons: an electrophysiological study in vitro.

Intracellular current-clamp recordings were performed using in vitro brainstem slice preparations to compare the actions of substance P, neurokinin A, neurokinin B and their agonists on rat dorsal vagal nucleus neurons with or without antagonists of neurokinin 1 and 2 receptors. The agonists used were either [Sar9,Met(O2)11]substance P or septide for neurokinin 1 and [Nle10]neurokinin A(4-10) for neurokinin 2 receptors. The antagonists were spantide, SR 140333 or RP 67580 for neurokinin 1 receptors and SR 48968 for neurokinin 2 receptors. Identification of vagal neurons was achieved electrophysiologically by testing antidromic responses and confirmed morphologically by an intracellular injection of biocytin. Of the 70 neurons tested, substance P led to depolarization in 36, hyperpolarization in six and no effect in 28. Depolarization was concentration dependent and generally associated with an increase of the membrane input resistance. Addition of tetrodotoxin (1 microM) to the medium had no effect on depolarization. RP 67580 (1 microM) blocked depolarization, but spantide and SR 140333 (microM to 50 microM) did not. Hyperpolarization was never observed using agonists. Neurokinin A and neurokinin 2 agonist induced concentration-dependent depolarization associated with an increase in membrane input resistance in eight of 14 neurons and in four of nine neurons, respectively. Depolarization was only partially abolished by the neurokinin 2 antagonist SR 48968. Neurokinin B had no effect in any of the eight neurons tested. These data prove that vagal neurons have neurokinin 1 and 2 receptors and that tachykinin could produce either depolarization or hyperpolarization. Since membrane potential variations were associated with an increase (during depolarization) or decrease (during hyperpolarization) in the membrane input resistance and since the reversal potential was close to the potassium equilibrium potential, we speculate that these effects are mediated by modulation of potassium conductance.

Action Potentials↗
PubMed1995

A-current modifies the spike of C-type neurones in the rabbit nodose ganglion.

1. In the rabbit nodose ganglion, C-type fibre neurones (C neurones) can be divided into two subtypes according to their after-hyperpolarizing potential (AHP) i.e. those with a fast AHP only and those with a fast AHP and a subsequent slow AHP produced by a slow calcium-dependent potassium current. In addition we have shown that some C neurones can be divided into two groups according to the effect of membrane hyperpolarization on their spikes i.e. type 1 in which duration and amplitude do not change and type 2 in which duration and amplitude decrease significantly. 2. In the present report we studied the effect of A-current (IA) on spike duration, amplitude and slow AHP using intracellular recording techniques. 3. To detect the presence of IA, we first applied a series of increasing rectangular hyperpolarizing pulses to remove IA inactivation and then a short depolarizing pulse to trigger a spike. In type 1 C neurones the lag time of the spike in relation to hyperpolarization remains constant whereas in type 2 C neurones the spike only appears after IA inactivation and lag time in relation to hyperpolarization is lengthened. Thus, type 2 C neurones have an IA while type 1 C neurones do not. The fact that addition of cadmium did not change the lag time in type 2 C neurones shows that the IA is not calcium dependent. 4. Nodose neurones can be orthodromically activated by stimulation of the vagal peripheral process. In this way, after a hyperpolarizing pulse, IA can be fully activated by the orthodromic spike itself. Under these conditions it is possible to analyse the effects of IA on the spike. This was done by increasing either the hyperpolarizing potential, pulse duration, or the delay of the spike after the end of the pulse. We observed that maximum IA inactivation removal was always associated with the lowest duration and amplitude of the spike. 5. When IA inhibitors, 4-aminopyridine (4-AP) or catechol, were applied to type 2 C neurones, the delay of the spike after the hyperpolarization-depolarization test was no longer observed. In addition 4-AP abolished the shortening of the duration of the spike induced by steady hyperpolarization. 6. In type 2 C neurones with slow AHP, the IA-related decrease in spike duration was associated with a disappearance of the slow AHP. This indicates that IA decreases the calcium influx during the spike.(ABSTRACT TRUNCATED AT 400 WORDS)

4-Aminopyridine↗
PubMed1995

Effects of substance P on cardiovascular regulation in the rabbit.

The effect of substance P on blood pressure and aortic reflex was investigated in rabbits. Microinjections of substance P and Sar9, Met(O2)11-SP (a selective NK1-receptor agonist) into the floor of the fourth ventricle led to a dose-dependent increase of blood pressure and a sharp enhancement of the baroreflex. These effects were abolished by pretreatment with SR 140333 (a selective NK1-receptor antagonist). Intraventricular injection of the antagonist alone significantly decreased the amplitude of the aortic reflex. After bivagotomy, the amplitude of the parasympathetic component of the baroreflex decreased dramatically and substance P injections were no longer effective. Our results demonstrate that substance P activation of NK1 receptors plays a major role in the modulation of the parasympathetic component of the baroceptor reflex.

Animals↗
PubMed1995

A new interface chamber for the study of mammalian nervous tissue slices.

We describe a new interface-type chamber for electrophysiological studies in mammalian brain slices. Thermoregulation of the inner recording chamber is achieved using the Peltier effect and a feedback control unit. Between 15 and 40 degrees C, and for perfusion rates from 1 to 5 ml/min, the temperature can be maintained within +/- 0.1 degrees C of the command value; it can also be rapidly and reliably changed. An external bath, heated by a coiled resistor, generates a humidified, oxygenated atmosphere diffusing above the slices. Survival of neuronal tissue is excellent and stable intracellular recordings can be obtained using either sharp or patch-clamp micropipettes. Perfusion solutions can be readily exchanged, rendering this chamber suitable for the study of bath-applied neuroactive compounds.

Animals↗
PubMed1994

Induction of Fos-like protein in neurons of the medulla oblongata after electrical stimulation of the vagus nerve in anesthetized rabbit.

Antibodies against the c-fos protein product Fos were used to map the first- and higher-order neurons in the rabbit medulla oblongata after electrical stimulation of the vagus nerve. Fos immunoreactivity appeared bilaterally except in the nucleus tractus solitarii. Seven areas were labeled: the nucleus tractus solitarii, the area postrema, the subnucleus lateralis caudalis magnocellularis medullar oblongata, the lateral reticular nucleus, the ambiguus nucleus, the dorsal part of the spinal trigeminal nucleus, the nucleus reticularis lateralis, the lateral border of the external cuneatus nucleus, the medial part of the inferior olivary nucleus (subnucleus beta). The last two areas have never been visualized with conventional tracing techniques and may represent higher-order neurons connected to visceral vagal pathways. No labeling was observed in the nodose ganglion.

Anesthetics↗
PubMed1993

Spike conduction properties of T-shaped C neurons in the rabbit nodose ganglion.

The electrical activity of C-type neurons was recorded intracellularly in the rabbit nodose ganglion maintained in vitro. The initial segment of their axon is spirally wound close to the cell body and a primary branching point divides it into a central process (CP) projecting to the nucleus of solitary tract in the medulla oblongata and a peripheral process (PP) which conveys sensory inputs from the viscera. Stimulation of the CP induced either somatic ("S") spikes or low-amplitude axonal ("A") spikes ("A1" or "A2"). In some cases abrupt changes in the latency of "S" or "A" spikes (jumps) were observed by gradually increasing the stimulus intensity. They are discussed in relation to a secondary branching on the central axon located inside or near the ganglion. Collision experiments showed that antidromic "A" spikes are blocked at the primary bifurcation of the axon (T-shaped neuron). Stimulation of the PP induced either "S" spikes or high amplitude "A" spikes ("A3" or "A4"). Orthodromic spikes could be blocked either before or after the primary bifurcation. When blocking occurs after the bifurcation on the stem axon, the spike can invade the central axon without invading the soma. The study of the refractory periods of the two processes and the application of high frequency stimulation showed that the PP allows higher frequencies than the soma and the CP, and thus that branching and the CP act as low-pass filters. These data support the view that the primary branching point and the CP of these T-shaped cells represent a strategic area to modulate visceral afferent messages.

Action Potentials↗
PubMed1991

Ultrastructural organization of the interstitial subnucleus of the nucleus of the tractus solitarius in the cat: identification of vagal afferents.

This electron microscopic study, based on serial section analysis, describes the synaptic organization of the interstitial subnucleus of the nucleus of the solitary tract and identifies the terminals of the vagal primary afferents utilizing degeneration and HRP transport. The interstitial subnucleus contains sparsely scattered cell bodies, numerous dendrites and axon terminals, and bundles of unmyelinated and myelinated axons. The cell bodies which are small in diameter have an organelle poor cytoplasm and a large invaginated nucleus. Axon terminals can be classified into two main types according to their vesicular shape. The first type contains clear, round vesicles and can be further subdivided into two subgroups on the basis of their morphology and the size of their vesicles. In the first subgroup the terminals are small, contain a few mitochondria and their vesicles are densely packed with an homogeneous size. In the second subgroup the terminals which vary from small to large, contain many mitochondria and contain round vesicles which are heterogeneous in size. The second main terminal type consists of axon terminals containing pleomorphic vesicles which are associated with asymmetrical or symmetrical synaptic contacts on dendrites. Axo-axonic contacts are present in the interstitial subnucleus. In general, the presynaptic axon terminals contain pleomorphic vesicles and the postsynaptic elements contain round vesicles of varying size. In some dendrites, identified by the presence of ribosomes, groups of round and/or pleomorphic vesicles are found associated with synaptic contacts. These dendrites are presynaptic to conventional dendrites and postsynaptic to axon terminals. After removal of the nodose ganglion, degenerative alterations are seen only at the caudal and middle levels of the interstitial subnucleus. Degeneration occurs in a few myelinated axons and in axon terminals which usually contain a mixture of small and larger round, clear vesicles. After HRP injection into the vagus nerve, the HRP reaction product is visible in axon terminals filled with clear, round vesicles which are heterogeneous in size. The labelled axon terminals establish single or multiple synaptic contacts. This study demonstrates that terminals of vagal primary afferents consist principally of terminals of the second subgroup. The morphology of these terminals are compared to primary afferents in the brainstem and spinal cord.

Afferent Pathways↗
PubMed1990

Serotonergic projections from the nodose ganglia to the nucleus tractus solitarius: an immunohistochemical and double labeling study in the rat.

Possible projections of serotonin (5-HT)-immunoreactive neurons in the nodose ganglia (NG) to the nucleus tractus solitarius (NTS) were investigated in the rat using a double labeling method combining retrograde transport and 5-HT immunohistochemistry. After injection of a complex of colloidal gold-apo-horseradish peroxidase into the medio-caudal and commissural parts of the NTS, most of the 5-HT-immunoreactive neurons were found to be labelled by the gold complex. The present study provides direct evidence for the existence, in the rat, of a serotonergic NG-NTS system. This system may be involved in the regulation of blood pressure and vigilance states.

Animals↗
PubMed1989

Electrophysiological and morphological properties of type C vagal neurons in the nodose ganglion of the cat.

Some passive and active electrical properties of type C neurons were studied intracellularly, in situ, in the nodose ganglia of adult cats. From the neuronal responses to hyperpolarizing and depolarizing rectangular current pulses it was possible to determine the input resistance (34.4 M omega) and specific membrane resistance (2373 omega.cm2). Significant changes in magnitude and duration of the action potential evoked by vagal stimulation result from changes in the resting potential caused by the passage of steady polarizing currents across the cell membrane. The action potentials evoked by infranodose vagal stimulation had a long duration, a long latency and comprised several components. The fast main spike was followed by a long post-hyperpolarization. The double shock technique showed that the fast main potential was composed of an initial segment spike ('A spike') and a somatic spike ('S spike'), and made it possible to determine the somatic refractory periods. After electrical identification of the cells, horseradish peroxidase was injected ionophoretically into the soma, and it was shown that the central processes were about four times smaller in diameter than the peripheral processes.

Action Potentials↗
PubMed1986

Anti-5-hydroxytryptamine antibodies: studies on their cross-reactivity in vitro and their immunohistochemical specificity.

Antibodies to 5-hydroxytryptamine (5-HT) were obtained from 4 rabbits after injections of 5-HT coupled to bovine serum albumin by means of paraformaldehyde (PF). Two methods were used to monitor the development of antibodies (AB): the one based on the "in vitro" competitive binding properties of the antibodies with 3(H)5-HT, the other, on their "in situ" binding properties to endogenous 5-HT, using the peroxidase-anti-peroxidase immunohistochemical technique, applied to paraffin embedded sections of cat brainstem. No pharmacological processing, detergents or proteolytic enzymes were used. The specificity of the antiserum was tested by competitive procedures with 20 analogs using the "in vitro" and "in situ" techniques. "In vitro" studies were performed with 5-HT free analogs and with analogs previously coupled with PF to lysine. Radioimmunological tests showed that the antibodies recognize mainly the ethylamine (CH2-CH2-NH2)-chain of the free analogs and that the best specificity was obtained with the 5-HT conjugate (5-HT-lysine-PF). The results suggest that the hapten is coupled through the phenolic positions C4 or C5. The "in situ" immunohistochemical extinction assays also revealed a distinct specificity for 5-HT. Possible optical and ultrastructural applications are illustrated in the raphé nuclei of the cat. These results confirm the reliability of radioimmunological tests for studying the specificity of AB directed against haptens, provided that haptens and analogs tested were first chemically transformed to resemble the immunogen (herewith lysine-PF coupling) with regard to its antigenic structure.

Animals↗
PubMed1986

[Nucleus tractus solitarius, serotonin and regulation of vigilance].

A brief historical survey of the vagal influences on sleep is lightly sketched first by analyzing the hypnogenic effects of the medullar area of the solitary tract nucleus (STN) where the vagal afferents terminate. Then the vagal effects and more extensively the sinusal or aortic effects are developed. It is shown that serotonin could play a crucial role in this so-called "vago-aortic" sleep mechanisms. Finally, these results are discussed and confronted with other experimental and human pathological data.

Animals↗
PubMed1983

Quantitative autoradiographic studies of 5-HT-accumulating neurones in the nodose ganglia of the cat after perikaryal or terminal uptake.

The distributions of ganglionar cell bodies, specifically labeled with 3H-5-HT, were studied in light microscope autoradiographs of the cat nodose ganglion. The size, number and localization of labeled cells were examined under different experimental conditions: after in vitro incubation of the ganglion with 3H-5-HT and after retrograde transport of 3H-5-HT injected into the nucleus of the solitary tract (NST) which receives primary visceral sensory projections from nodose ganglia cells of the vagal nerve. Following incubation of the nodose ganglia with a low concentration of 3H-5-HT (10(-6) M), some ganglionar cell bodies took up and retained the tracer. In both the right and left ganglia, they were significantly smaller in size than the unreactive neurones. The mean diameter of their perikaryon was 36.97 +/- 0.52 microns, compared with 45.76 +/- 0.87 microns in unreactive neurones. About 600 labeled cell bodies were counted in each ganglion, corresponding to 2-3% of the total nodose ganglion cell population. These reactive neurones were not localized in one particular area of the ganglia, but scattered throughout both of them. Following bilateral or unilateral microinjections of 3H-5-HT in the NST, retrogradely labeled cell bodies were observed, 24 h later, in the nodose ganglia. Their mean diameter was estimated to be 36.14 +/- 0.69 microns and they represented approximately 2% of the total ganglion cell population. As in in vitro experiments, the labeled cells were not grouped in any particular region of the ganglion. These experiments show that the distribution of both populations of labeled cells, observed under these conditions, are comparable. On an anatomical and quantitative basis one may reasonably suppose that the perikaryal or terminal uptakes concern the same neuronal population.

Animals↗
PubMed1983

Simultaneous visualization of aortic and [3H]5-hydroxytryptamine-accumulating cell bodies in the nodose ganglion of the cat.

1. Single- and double-tracer experiments were performed in cats to investigate the relationship between the aortic cells and the cell bodies accumulating 5-hydroxytryptamine (5-HT) in the nodose ganglion. In one series of experiments, horseradish peroxidase (HRP) was applied to the central end of the aortic nerve, anterogradely transported and accumulated in ganglionar perikarya. The distribution of HRP-positive neurones was reconstructed in serial sections through the nodose ganglion. In a second series of experiments, the distribution of [(3)H]5-HT-accumulating cell bodies was assessed following incubation of the nodose ganglion in [(3)H]5-HT. The third series of experiments combined the treatments of the preceding ones: anterograde transport of HRP in the aortic nerve followed by incubation of the nodose ganglion in [(3)H]5-HT.2. The results from these experiments provide more information with regard to (i) the topographical relationship between the aortic and [(3)H]5-HT-accumulating cell bodies in the same ganglion and (ii) the distribution and number of double-labelled neurones, giving further indications about histochemical components of the aortic nerve.3. The HRP experiments demonstrated that HRP-positive cells show a preferential pattern of topographical organization. They were mostly located in the medial border of the ganglion where the laryngeal and aortic nerves enter. On the other hand, [(3)H]5-HT-accumulating neurones were scattered throughout the ganglion.4. In double-tracer experiments, three populations of labelled cell bodies were distinguished in the same nodose ganglion: (1) single HRP-cells; (2) single [(3)H]5-HT-accumulating cells and (3) double-labelled cells. The distribution of the latter population exhibited no preferential localization. Quantitative estimates indicated that double-labelled neurones constituted 65-85% of the population of HRP-positive cell bodies.5. These results show that most aortic neurones are able to take up exogenous serotonin and may be serotonergic neurones. They suggest that serotonin may be involved in physiological effects mediated via the aortic nerves.

Animals↗
PubMed1982

Serotonin-like immunoreactivity in paraffin-sections of the nodose ganglia of the cat.

The presence of serotonin (5-HT)-like immunoreactivity has been detected in ganglionar cell bodies of the nodose ganglia of the cat, using the peroxidase-antiperoxidase immunohistochemical method, applied to sections of ganglia previously embedded in paraffin. The antibodies were raised in rabbits following injection of 5-HT conjugated to bovine serum albumin, with formaldehyde as the condensation reagent. Immunoreactive cell bodies are morphologically and topographically described. The ubiquitous distribution and non-uniform labelling suggest that these 5-HT-containing vagal neurones may give rise to several functionally different 5-HT types in vagal afferent fibres.

Animals↗
PubMed1982

Selective retrograde transport of 3H-serotonin in vagal afferents.

A new serotonergic afferent vagal component has been demonstrated in the cat by radioautography. Twenty-four hours after a bilateral injection of tritiated serotonin (3H-5-HT) into the area of the nucleus of the solitary tract (NST), heavily and lightly labelled cell bodies were observed in the nodose ganglia. After unilateral injections of 3H-5-HT into the same area, labelled ganglionar cell bodies were found in the ipsilateral nodose ganglion. Some were also found in the contralateral one, suggesting a serotonergic crossed fibers component. Dense clusters of silver grains, depicting typical labelling of neuronal varicosities, were observed in the NST. After destruction of the serotonergic terminals with 5,7-dihydroxytryptamine, followed by injection of 3H-5-HT, the number of labelled cell bodies decreased dramatically in the ipsilateral nodose ganglia and the clusters of silver grains disappeared in the NST. After ligature or section of the supranodose vagal nerve, following injection of 3H-5-HT into the NST, no radioautographic reaction was observed in the homolateral nodose ganglia. The present study demonstrates the existence of a peripheral serotonergic system in vagal afferents. The physiological implications of this new serotonergic visceral pathway remain to be studied.

Afferent Pathways↗
PubMed1982

A single radioimmunological assay for serotonin, N-acetylserotonin, 5-methoxytryptamine, and melatonin.

A radioimmunoassay of melatonin using a new iodinated derivative has been developed. Simple chemical treatments have then been designed to convert serotonin, N-acetylserotonin, and 5-methoxytryptamine to melatonin. Thus these four molecules, belonging to the same metabolic pathway, were separately assayed in the same radioimmunological system at the same sensitivity level (0.01 pmol). Some biological results on the circadian variations of melatonin and serotonin in blood, pineal, and miscellaneous brain structures are presented.

5-Methoxytryptamine↗