Fabrice Wallois

c-fos-like immunoreactivity in the cat's neuraxis following moderate hypoxia or hypercapnia

The overall pattern of c-fos immunoreactivity was studied in the brainstem and spinal cord of cats subjected to moderate hypoxia or hypercapnia. In control cats (normoxic, normocapnic), c-fos was expressed mainly in pontine and periaqueductal grey but not in brainstem structures engaged in respiratory control nor in the spinal cord. Both hypoxia and hypercapnia induced c-fos expression in the parabrachial area (pneumotaxic center). In the retrotrapezoid nucleus, a structure involved in respiratory rhythmogenesis and chemoreception, immunoreactivity was detected in hypoxic but not in hypercapnic cats. Neurons in the nucleus raphe pallidus preferentially expressed c-fos in response to hypercapnia. Labelled neurons were concentrated in the dorsal and gelatinosus subnuclei of the solitary tract following hypoxia and hypercapnia, respectively. Our data suggest that some neurons that express c-fos in hypoxic or hypercapnic cats may be involved in coordination of cardiovascular and respiratory function.

Trigeminal nasal receptors related to respiration and to various stimuli in cats

In twenty adult cats of either sex under nembutal anaesthesia, we aimed at delineating the sensitive territory of trigeminal nerves innervating the nasal mucosa. The different trigeminal nerves (anterior ethmoidal, posterior nasal and infraorbital nerves) were dissected in the orbit. Activity of these nerves was recorded during spontaneous nasal and tracheal breathing and in response to various stimuli: mechanical (manual probing and air jets) and irritants (ammonia vapours). Multiple and unitary activity recorded in nerve filaments enabled a classification of the receptors on the basis of their discharge pattern as rapidly-, intermediately- or slowly adapting receptors, and as drive or non-drive nasal receptors depending on whether or not the respiratory modulation was preserved during tracheal breathing.

C-Fos-like immunoreactivity in the cat brainstem evoked by sneeze-inducing air puff stimulation of the nasal mucosa.

Sneeze is one of the most important protective reflex of the respiratory tract. It is elicited from trigeminal peripheral fields and results in major changes in the discharge patterns of the medullary respiratory-related neurons. The pattern of c-Fos-like immunoreactivity evoked by sneezing was explored as a structural approach to the networks involved in this particular model of trigemino--respiratory interactions. Sneezes were elicited in anaesthetized adult cats by driving air puffs to the superior nasal meatus through a catheter. Additional cats were used as controls for anaesthesia and for catheter insertion into the nostril. In sneezing cats, immunoreactivity was evoked in projection areas of the ethmoidal afferents which innervate the superior nasal meatus, e.g. the subnuclei caudalis, interpolaris and in the interstitial islands of the trigeminal sensory complex. Immunoreactivity was also markedly enhanced in the areas devoted to respiratory control in the medulla (solitary complex, nucleus retroambiguus) and in the pontine parabrachial area. C-Fos expression was also evoked in the lateral aspect of the parvicellular reticular formation in sneezing cats. This area might be of major importance in the adaptation of the ventilatory system to expulsive functions.

Autonomic response and Fos expression in the NTS following intermittent vagal stimulation: Importance of pulse frequency

Chronic intermittent stimulation of the vagus nerve (VNS) is an approved adjunctive therapy of refractory epilepsy. Nevertheless, the circuits triggered by VNS under the variable conditions used in patients are not well understood. We analyzed the effect of increasing pulse frequency on physiological variables (intragastric pressure, cardiac and respiratory frequencies) and neuronal activation in the solitary tract nucleus (NTS), the entry level of peripheral vagal afferents, in the rat. For this purpose, we compared the subnuclear distribution of Fos-like immunoreactivity within the NTS following VNS at frequencies selected for their low (1 Hz) or high (10 Hz) therapeutic efficacy. In addition, NADPH diaphorase histochemistry was conducted in double-labeling experiments to check whether activated neurons may express nitric oxide (NO). We demonstrated that increasing pulse frequency had a major influence on the cardiorespiratory response to VNS and on the amount of activated neurons within NTS subdivisions engaged in cardiorespiratory control. These data, in line with clinical observations, suggested that within the range of therapeutic frequency, VNS may favor the regulation by vagal inputs of cortical activities within limbic areas involved in both epileptogenesis and cardiorespiratory afferent control. Furthermore, we did not find any evidence that anticonvulsant VNS might trigger NOergic neurons in the NTS.

Influence of trigeminal nasal afferents on bulbar respiratory neuronal activity.

This study examined the influence of nasal trigeminal afferents, the anterior ethmoidal nerve (AEN) and posterior nasal nerves (PNN) on the spike discharges of respiratory-related neurons recorded in the ventral respiratory group (VRG) (2.6-3.5 mm lateral to the midline, from 1 mm rostral to 3 mm caudal to the obex and at depth of 2-4 mm below the dorsal surface). Electrical stimulations to the AEN and PNN were administered to 10 pentobarbital anaesthetized cats and to 8 ketamine anaesthetized, vagotomized, curarized and ventilated cats. Single shock stimulations of either nerve evoked transient and total inhibition of inspiratory activities. Expiratory-related neurons of the VRG presented three patterns of activity in response to stimulation:excitation, inhibition or inhibition followed by excitation. More generally, expiratory units are activated with a short latency. In the course of repetitive stimulation of the AEN and PNN we observed a prolongation of the spontaneous inspiratory discharge which presented transient, short inhibition in response to each shock. Most expiratory units presented a short activation which was synchronous with the transient inhibition of inspiratory activities. When repetitive stimulation provoked a sneeze-like response, we observed a progressive increase in the duration of transient inspiratory inhibition first, associated with a progressive reinforcement of transient expiratory activation. Secondarily, just before the expiratory thrust, we noted a stronger inhibition of the inspiratory activity which preceded a high-frequency (400 Hz) expiratory discharge. Nasal afferents exert a forceful excitatory effect on bulbospinal (BS) and non-bulbospinal-non-vagal (NBS-NV) expiratory cells of the VRG. The effects due to vagotomy and curarization are discussed.

Trigeminal afferences implied in the triggering or inhibition of sneezing in cats

The aim of this study was to precise the role of the different trigeminal nerve branches involved in innervation of the nostril in triggering the sneeze reflex. Electrical stimulation of the anterior ethmoidal, posterior nasal and infraorbital nerves was performed in anaesthetized cats. Stimulation of these 3 nerves produced sneezing identical to that induced by mechanical stimulation. Our results emphasize inhibition of the sneeze reflex related to stimulation of the anterior ethmoidal or the posterior nasal nerves by stimulation of the infraorbital nerve.

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.

Postnatal development of the anterior ethmoidal nerve in cats: unmyelinated and myelinated nerve fiber analysis.

This is the first quantitative electron microscopic study of anterior ethmoidal nerve in adult and newborn cats. The adult nerve comprises about 1,000 myelinated fibers including A delta (65%) and A beta (35%) fibers and 6,000 unmyelinated fibers. At birth, only 27% of the adult myelinated fibers complement is already present. The immaturity of the nerve is discussed in relation to that of the sneeze reflex.

Nasal air puff stimulations and laryngeal, thoracic and abdominal muscle activities

In cats, we studied the activity of laryngeal, thoracic and abdominal muscles and the variations in oesophageal pressure in response to air puff stimulations of the nasal mucosa. Following single stimulations, inspiratory and laryngeal dilator muscles were transiently inhibited. During inspiratory inhibition, expiratory muscles and laryngeal constrictor were transiently activated. Repetitive air puff stimulations, which induced sneeze, evoked a similar pattern of transient activities during the inspiratory preparation of sneeze. This resulted in transient fluctuations of the oesophageal pressure, whose mean value became more negative as the preparatory inspiration enhanced. Our results suggest that the entire pool of respiratory neurons (bulbospinal, vagal and facial) works together in the sneeze reflex. Study of transient inspiratory inhibition demonstrates two periods during the preparatory inspiration phase of sneeze. In the first period transient effects are related to each shock of the stimulation. The second period is characterized by a diminution or a lack of transient effects associated with the stimulation. At the end of the expulsive phase, the diaphragm and the glottal dilator were further activated.

Influence of vagal afferents in the sneeze reflex in cats

We studied the effects of bilateral vagotomy and step pulmonary inflations (5, 10, 15 mmHg, i.e., 0.66, 1.33, 2 kKPa) on sneeze reflex in ketamine-anaesthetized cats. Bilateral vagotomy lengthens the duration of preparatory inspiration and diminishes the amplitude of expiratory activities in sneeze. In contrast, 5 mmHg pulmonary inflation facilitates the sneeze. It shortens the inspiratory preparation and increases the frequency of sneeze attacks. At 10 mmHg pulmonary inflations, inspiration is inhibited and only expiratory thrust occurs. At 15 mmHg pulmonary inflations, vagal afferent stimulations inhibit the sneeze.