Connections between retrotrapezoid nucleus and nucleus tractus solitarii in cat.

The retrotrapezoid nucleus (RTN), a part of the rostral ventrolateral medulla, is involved in the control of breathing. The mechanisms by which the RTN modulate the activity of respiratory neurons during chemoreceptor stimulation are not fully understood. This electrophysiological study performed in the cat demonstrates that 18 out of 22 RTN neurons receive inputs from the commissural subnucleus of the solitary tract (cNTS), the peripheral chemoreceptor afferents projection site. Moreover, six RTN neurons are found to present interconnection between RTN and the ventrolateral subnucleus of the solitary tract, an area containing mainly bulbo-spinal respiratory neurons. The present data suggest that RTN play a key role by concomitantly integrating chemosensitive informations relayed by the cNTS and providing an influence on the respiratory network.

Effect of hypoxia on the activity of respiratory and non-respiratory modulated retrotrapezoid neurons of the cat.

The retrotrapezoid nucleus (RTN), a part of the rostral ventrolateral medulla, is involved in the control of breathing. A recent immunohistological study suggested a possible involvement of the RTN in hypoxic chemoreflex loop. The present electrophysiological study performed in the cat demonstrates that 23 out of 24 RTN neurons were stimulated during the biphasic respiratory response to hypoxia, which consists of a reinforcement followed by a depression of respiratory activity. This confirms the previous immunohistological study. While 15 RTN neurons might integrate either phase I (n = 7) or phase II (n = 8) O2-chemosensitive inputs, the remaining eight RTN neurons stimulated by hypoxia are susceptible to integrate both phase I and phase II O2-chemosensitive inputs. In conclusion, our results suggest that the different subsets of RTN neurons may influence respiratory output by conveying signals originating from peripheral and/or central chemoreceptors stimulated during hypoxia.

Role of respiratory and non-respiratory neurones in the region of the NTS in the elaboration of the sneeze reflex in cat.

Extracellular recordings were made in the dorsal respiratory group (DRG) and adjacent reticular formation following single-shock stimulation of the anterior ethmoidal nerve (AEN) and during sneeze evoked by repetitive stimulation of the AEN in nembutal-anaesthetized, curarized and ventilated cats. These neurones were characterised according to (i) their activity during the respiratory cycle (as inspiratory augmenting or decrementing (I Aug or I Dec), expiratory augmenting or decrementing (E Aug or E Dec), silent or tonic), and (ii) their axonal projection (bulbospinal or non-bulbospinal-non-vagal (BS or NBS-NV)). Following single-shock stimulation of the AEN, most of the inspiratory neurones were transiently inhibited, whereas E Aug neurones were activated and E Dec neurones were activated and then inhibited. Silent neurones responded with a multispike or a paucispike pattern. Following repetitive stimulation of the AEN and during the resulting sneeze reflex, I Aug neurones increased their activity in parallel with the phrenic activity, I Dec neurones fired at the onset and at the end of the inspiration, E Dec and some silent neurones fired either during the compressive phase or after the expulsive phase, whereas E Aug and some silent neurones fired during the expulsive phase. We conclude that sneeze involves a reconfiguration of the central respiratory drive which uses, at least partly, the respiratory network to trigger a non-ventilatory defensive motor act.

[The inhibitory action of a saccharide derivative on a calcium channel in the oocyte of Xenopus]. / Mise en évidence de l'action inhibitrice d'un dérivé saccharidique sur le canal calcique de l'ovocyte de Xénope.

Endogenous calcium channels of Xenopus oocyte membrane do not fit with pharmacological classification of calcium channels. The present study demonstrates that the saccharidic derivate, OC8-MAGlu-MAGlu, has potent inhibitory effect on this channel activity.

Interaction between Ca2+ release from inositol trisphosphate sensitive stores and Ca2+ entry through neuronal Ca2+ channels expressed in Xenopus oocyte.

Rat cerebellar RNA injected into Xenopus oocytes leads to the expression of putative P-type voltage-dependent Ca2+ channels (VDCCs). The monitoring of intracellular Ca2+ variations by recording the Ca2+ dependent chloride current in voltage clamped oocytes indicates that activation of these Ca2+ channels by depolarization gives rise to two distinct components of cytosolic Ca2+ elevation. If the early component (T1) can be directly attributed to the Ca2+ entry through VDCCs, the second delayed one (T2) is related to a Ca2+ release from InsP3 sensitive stores activated following Ca2+ entry. Modifications of cytosolic Ca2+ by direct injection of Ca2+ into oocytes or by increasing the Ca2+ influx through VDCCs suggest that the Ca2+ release from intracellular InsP3 sensitive stores can be modulated in a differential manner. Namely, discrete elevations of cytosolic Ca2+ switch on the Ca2+ release whereas higher Ca2+ concentrations dampen the release. These results suggest a functional coupling between P-type VDCCs and InsP3 receptors.

[Effects of the monosaccharide derivative 8RN-DAGal on the putative P-type calcium channel expressed in Xenopus oocytes]. / Effets du dérivé monosaccharidique 8RN-DAGal sur un courant calcique apparenté au type P exprimé dans l'ovocyte de Xénope.

P-type calcium channels are expressed in Xenopus oocytes after injection of rat cerebellar mRNA. The FTX and omega-Aga-IVa toxins extracted from Agelenopsis aperta venom are known to inhibit the activity of this channel. The present results demonstrate that 8RN-DAGal is also a antagonist of P-type calcium channels. The inhibition of the current, obtained with Ba2+, as charge carrier, is voltage dependent.

Positive regulation by protein kinase C of slow Na current in Xenopus oocytes.

The slow inward Na current observed during sustained depolarization of the Xenopus oocyte membrane is due to a complex mechanism described as the induction of the channels. The present work investigates the role of protein phosphorylation in Na channel function. Injection of alkaline phosphatase in the oocytes decreased inward current. Therefore, the possible involvement of protein kinase in Na channel induction was explored. Treatment of oocytes with two activators of protein kinase C (PKC) resulted in enhanced Na current amplitude, whereas the treatment of oocytes with two potent PKC inhibitors decreased the inward current. These results imply that PKC phosphorylation is a fundamental step of Na channel induction. The possibility that the depolarization of the oocyte membrane may be the factor involved in PKC activation is discussed.

[Structure-activity relationship of a new family of calcium antagonist molecules derived from monosaccharides]. / Relations structure-activité d'une nouvelle famille de molécules antagonistes calciques derivées de monosaccharides.

The Ca++ antagonist effects of new drugs derived from monosaccharides were tested in the rat duodenal muscle preparation in vitro. The structure-activity relationship shows that: 1. The monoacetonide products (nR-O-MAG and nR-S-MAG) with alkyl chain from 8 to 9 C atoms induce a maximal reduction of muscular tonus and contraction. 2. The inhibitory effect rapidly decreases when the alkyl chain has a number of C atoms smaller than 7 or larger than 9. 3. The diacetonide (DAG) and acetonide (G) products induce an inhibition of less extent. 4. The Ca++ antagonist effect is very slightly changed by the type of heteroatom (O or S) linking the alkyl chain to the monosaccharide. 5. The type of monosaccharide affects the Ca++ antagonist activity.

Regulation by protein kinase-C of putative P-type Ca channels expressed in Xenopus oocytes from cerebellar mRNA.

Xenopus oocytes injected with rat cerebellar mRNA expressed functional voltage-dependent Ca channels detected as an inward Ba current (IBa). The pharmacological resistance to dihydropyridines and omega-conotoxin together with the blockade obtained with Agelenopsis aperta venom suggest that these channels could be somehow assimilated to P-type Ca channels. The precise nature of the transplanted Ca channels was assessed by hybrid-arrest experiments using a specific oligonucleotide antisense-derivated from the recently cloned alpha 1-subunit of P channels (BI-1 clone). In addition, we demonstrate that exogenous Ca channel activity was enhanced by two different PKC activators (a phorbol ester and a structural analog to diacylglycerol). The general electrophysiological and pharmacological properties of the stimulated Ca channels remain unchanged. This potentiation induced by PKC activators is antagonized by a PKC inhibitor (staurosporine) and by a monoclonal antibody directed against PKC. It is concluded that P-type Ca channels are potentially regulated by PKC phosphorylation and the functional relevance of this intracellular pathway is discussed.

Somatosensory evoked potentials after median and tibial nerve stimulation in healthy newborns.

Cortical and spinal somatosensory evoked potentials (SEPs) have been recorded after median and tibial nerve stimulation in healthy newborns. Spinal SEPs were readily obtained and recorded in all but one neonates after stimulation of both nerves. Cortical SEPs were more frequently recorded after median nerve (87%) than after tibial nerve stimulation (73%) but the shape of cortical SEPs obtained after tibial nerve stimulation was less variable. The mean feature of cortical SEPs was a negative wave (N27) for median nerve and a positive wave (P32) for tibial nerve. The present results demonstrate the feasibility of obtaining in the same baby, spinal and cortical SEPs after stimulation of median and tibial nerve, giving information on the functional integrity of central and peripheral somatosensory pathways which supply upper and lower limbs.

Phrenic-to-phrenic inhibition and excitation in spinal cats.

In decerebrate, C2-spinalized cats, stimulation of the C6-phrenic root produces a weak activation of phrenic motoneurons in the adjacent C5 segment in a few animals (23%). When phrenic motoneurons are electrically excited by testing stimulation applied to the spinal cord or internal intercostal nerve, the evoked responses recorded in a cervical phrenic root are partly inhibited by conditioning stimulation applied to another ipsilateral or contralateral cervical phrenic root. We therefore conclude that phrenic fibers exert both inhibitory and excitatory effects on adjacent phrenic motoneurons in the cervical spinal cord.

Effects of ipsilateral and contralateral cervical phrenic afferents stimulation on phrenic motor unit activity in the cat.

The phrenic-to-phrenic inhibitory reflex has been analyzed using recordings of activity of single C5-phrenic motor units (PMUs). After ipsilateral or contralateral stimulation of the C6-phrenic root, early and late PMUs exhibit a similar inhibition. After contralateral stimulation, the duration of the inhibition is smaller and the threshold stimulus is higher than the corresponding values observed after ipsilateral stimulation. The latency of the inhibition is similar for both stimulations. Hemispinalization, contralateral to the recording site, affects weakly the phrenic-to-phrenic reflex. We conclude that early and late PMUs receive a similar inhibitory input from phrenic afferents and that the inhibition observed after cervical phrenic nerve stimulation may involve spinal cord circuits.

[Conduction velocities of motor, cutaneous sensory, and IA proprioceptive fibers in normal newborn infant at term]. / Vitesses de conduction des fibres motrices, sensitives cutanées et proprioceptives IA chez le nouveau-né normal à terme.

Conduction velocities of non-nociceptive cutaneous and IA afferent fibres and alpha-motor fibres from ulnar, median, deep peroneal, posterior tibial and sural nerves have been measured in healthy full-term newborns, using surface electrodes. There is an overlapping in the distribution of the individual values of conduction velocities for the different types of fibres. The conduction velocities of cutaneous afferent fibres are not different in the upper or lower limbs. The conduction velocities of motor and IA fibres are higher in the upper than in the lower limb. In the upper limb, conduction velocities of IA fibres are higher than those of motor fibres, as they are in the lower limb for the posterior tibial nerve. Moreover, conduction velocities of motor fibres do not differ in males and females.

Inhibitory and excitatory effects on respiration by phrenic nerve afferent stimulation in cats.

Respiratory effects of electrical stimulation of phrenic nerve afferents were studied in anesthetized cats, either spontaneously breathing or paralyzed and ventilated. The type of phrenic afferent fibers activated was controlled by recording the evoked action potentials from dorsal root fibers. In both preparations, stimulation at a strength sufficient to activate small diameter myelinated phrenic nerve afferents induced a biphasic response. The first phase lasted a few respiratory cycles and was inhibitory and consisted of a decrease in tidal volume (VT) or phrenic activity (NA), inspiratory time (TI), respiratory duty cycle (TI/Ttot) and instantaneous ventilation (VE) or minute phrenic activity (NMA). Expiratory time (TE) increased and breathing frequency (f) and mean inspiratory flow (VT/TI) or mean inspiratory neural activity (NA/TI) did not change. This short-term response was suppressed in animals pretreated with bicuculline. The second phase was a long-term excitation in which VT or NA, f, VE or NMA and VT/TI increased whereas both TI and TI/Ttot decreased and TE did not change. Unlike published data, our results suggest that small-diameter myelinated phrenic nerve afferents are involved in these responses. These phrenic fibers, like afferents from other muscles, affect respiratory output and may play a role in the control of breathing.

The excitation by neurotensin of nucleus tractus solitarius neurons induces apneustic breathing.

The possible involvement of neurotensin in the regulation of respiratory drive has been tested on single brainstem respiratory related neurons and on the global respiratory output. The neuropeptide was locally applied either by microiontophoresis or by pressure injection in the dorsal and ventral respiratory areas of the anesthetized bivagotomized cat. Effects of neurotensin applications were studied, on the one hand on the firing discharge of respiratory related neurons and on the other hand on the phrenic nerve activity and on arterial blood pressure. An increase of the firing frequency of respiratory related neurons was induced by neurotensin applied by iontophoresis or by pressure injection (0.005-33.5 fmol/s) on single neurons. In the latter case, neurotensin was active at concentration 10(3) times lower than glutamate. A bilateral apneustic pattern was induced on the phrenic nerve activities by microinjection of neurotensin (0.23-0.54 pmol/s) in one ventrolateral nucleus tractus solitarius without alteration of arterial blood pressure. These results suggest that the release of neurotensin in the nucleus tractus solitarius regulates respiratory rhythmogenesis by increasing the inspiratory duration.

[Non-participation of bulbar inspiratory neurons in the esophago-diaphragmatic inhibitory reflex]. / Non-participation des neurones inspiratoires bulbaires dans le réflexe inhibiteur oesophago-diaphragmatique.

The effects of the distension of the lower oesophageal sphincter were studied on the inspiratory activity of 96 medullary neurons located either in the dorsal or in the ventral respiratory groups and on the inspiratory activity of the costal and crural parts of the diaphragm in barbiturate anaesthetized cat. Inhibition of the inspiratory activity of the crural part of the diaphragm during oesophageal distension was never associated with significant changes of the medullary inspiratory neuron discharge. These results suggest that the observed crural inhibition is due to reflex loop that does not include the inspiratory neurons belonging to the dorsal and the ventral respiratory groups.

Effects of stimulation of phrenic afferent fibers on medullary respiratory neurons in cat.

The effects of electrical stimulation of both cervical branches (C5 and C6) of the right phrenic nerve on medullary respiratory neuron activity were studied in anesthetized, spontaneously breathing cats. In 14 cats, the stimulation of the thin phrenic afferents had no effect on the inspiratory duration and evoked excitatory or inhibitory responses in only 3/86 inspiratory neurons tested. In 3 cats, the stimulation decreased the inspiratory duration and 26/26 inspiratory neurons showed a shortened discharge without modification of their discharge frequency. Although the effects of the stimulation were not analysed by averaging techniques, it is concluded that phrenic afferents do not exert an important control on the medullary respiratory neuron discharge.

Effects of tonic vagal input on breathing pattern in newborn rabbits.

Respiratory effects of positive and negative pressure breathing were studied in 1- and 4-day-old rabbit pups anesthetized with ketamine (50 mg/kg, im) and acepromazine (3 mg/kg, im). We recorded tidal volume (VT), tracheal pressure (Ptr), and integrated diaphragmatic EMG (DiEMG). Inspiratory (TI) and expiratory time (TE) were measured from the records of DiEMG. During breathing with increased Ptr by 1 or 2 cmH2O, VT, minute ventilation (VE), and respiratory rate (f) decreased. Changes in f relied on a TE prolongation. Neither DiEMG nor its rate of rise (DiEMGt) were affected. Except for VT decrease during positive Ptr, all other effects disappeared after vagotomy. Our results indicate that an increase in tonic vagal activity interacts with the mechanisms controlling TE and has no effect on depth and duration of inspiration. When Ptr decreased by 1 and 2 cmH2O, VE increased due to an increase in f. Increase in f relied on shortening of both TI and TE; the TE effect being more pronounced. DiEMG and DiEMGt also increased. Adverse effects of lung deflation and vagotomy strongly suggest that the respiratory reflex stimulation due to decrease in Ptr does not rely on inhibition of the slowly adapting stretch receptor activity. Therefore other excitatory vagal inputs must be responsible for this response. We propose two vagally mediated inputs: the irritant and/or the cardiac receptors.

Phrenic afferent input to the lateral medullary reticular formation of the cat.

The afferent inputs from phrenic nerve stimulation to the lateral reticular formation of the lower brain stem were studied in anesthetized spontaneously breathing cats. The activity of reticular neurons was recorded by means of extracellular tungsten microelectrodes. Electrical stimulation of the central end of the right phrenic nerve evoked excitatory or inhibitory responses in the lateral reticular nucleus (LRN), in the nucleus ambiguus (AMB) and in a region dorsal to the AMB of ipsi- and contralateral sides. Phrenic afferents belonging to the flexor reflex afferent group were involved in these responses. The discharge pattern of the respiratory related units (RRU) of the AMB were exceptionally affected by phrenic nerve stimulations. It is concluded that high threshold phrenic afferents relay in the LRN before projecting to the cerebellar cortex. The overlapping of respiratory and non-respiratory afferents in the reticular formation may participate to the adaptations of respiratory and somatomotor functions during specific behaviors.

Projection of phrenic afferents to the external cuneate nucleus in the cat.

The present study, performed on anesthetized, spontaneously breathing cats, deals with the projection of group I and II muscle afferents of the phrenic nerve (PN) to the external cuneate nucleus (ECN). Stimulation of the central end of the PN evoked a complex response in the ipsilateral ECN. Two principal components could be distinguished in this potential from the respective absolute refractory periods (ARP) and from the effect of antidromic stimulation in the ECN. Thus, the early group of waves may correspond to recordings of direct fibers and the later group to postsynaptic activations within the ECN. Similar to the forelimb nerves and intercostal nerves of the upper intercostal spaces, the larger muscle afferents of the PN project to the ECN.