Inspiratory on-switch evoked by mesencephalic stimulation: activity of medullary respiratory neurones

Summary The activity of medullary inspiratory and expiratory neurones was studied in urethan-chloralose anaesthetized cats during stimulus — evoked inspiratory phase (inspiratory on-switch). All neurones were characterized according to their axonal destination (i.e. bulbospinal neurones or vagal motoneurones) or the absence of such axonal projections (i.e. propriobulbar neurones), and to their location in the dorsal or ventral respiratory nuclei. 1. The inspiratory on-switch effects were elicited during expiration (E phase) by brief tetanic electrical stimulation (50 to 100 ms duration; 0.5 mA; 300 Hz) delivered to the mesencephalic periaqueductal central gray and the adjacent reticular formation. The evoked inspiratory effects observed on the phrenic nerve discharge consisted of: (i) an immediate response (latency 20 ± 5 ms) of stable duration related to the stimulus (primary response: Prim.R.), (ii) a delayed response (patterned response: Patt.R.) appearing after a latent period (silent phase: Sil.P.) of 100 ms maximal duration. The later the stimulus in the E phase, the longer was the duration of the Patt.R. (300 to 1000 ms). 2. The stimulation evoked an earlier activation of the inspiratory bulbospinal neurones (latency 12 ± 6 ms) than that obtained in the phrenic nerve (Prim.R.). Hence, the Prim.R. originated from the bulbospinal pathway and not from a pathway directly impinging on the motoneurones. Conversely during stimulation very few inspiratory propriobulbar neurones were activated and no expiratory neurone discharged. 3. During the phrenic Sil.P., 46% of the inspiratory bulbospinal neurones continued to discharge with a firing rate lower than that during the stimulus train, while most of the inspiratory propriobulbar and expiratory neurones were not active. 4. During the Patt.R. all inspiratory bulbospinal neurones discharged early and were strongly activated whatever the Patt.R. duration whereas the expiratory neurones were not active. Inspiratory propriobulbar neurones were either not recruited or recruited later, and the number of active neurones increased as the duration of the Patt.R. lengthened. 5. Our results suggest that the eliciting of the stimulus-evoked inspiration (Patt.R.) primarily depends on the activation of the inspiratory bulbospinal neurones. These neurones therefore would not only be the output neurones of the medullary respiratory centres, but they would serve other roles such as building up of the excitation in other respiratory neurones, thus acting as a component of the inspiratory ramp generator.Abbreviations Prim.R Primary response - Patt.R Patterned response - Sil.P Silent phase - I phase Inspiratory phase - E phase Expiratory phase - IBSN Inspiratory bulbospinal neurones - IPBN Inspiratory propriobulbar neurones - EBSN Expiratory bulbospinal neurones - EPBN Expiratory propriobulbar neurones - DRN Dorsal respiratory nucleus - VRN Ventral respiratory nucleus Supported by CNRS (LA 205 and ATP no 4188) and Fondation pour Ia recherche médicale     

Membrane potentials of individual cells of isolated gastric glands of rabbit

In urethane-anaesthetized, paralyzed and artificially ventilated rabbits, medullary respiration-related neurons (RRU) were classified according to the phase relation of their burst discharge to phrenic nerve activity. Phase-bound inspiratory (I) or expiratory (E) neurons were discriminated from phase-spanning expiratory-inspiratory (EI) or inspiratory-expiratory (IE) units. Mechanisms of termination of inspiration by electrical stimulation of rostral pontine nuclei (Nc. parabrachialis medialis; Lc. coeruleus) were examined firstly to demonstrate whether RRU receive descending excitatory and inhibitory afferents as well as ascending efferents and secondly to analyse the time course of the neuronal pathways involved. Of 120 RRU, 38 neurons were demonstrated to receive pontine afferents. About 33% of all E neurons became orthodromically excited during rostral pons stimulation whereas 18.2% of all I cells became orthodromically inhibited. Some RRU were shown to project up to the rostral pons. 50% of these were of the phase-spanning IE type. The onset of inspiratory inhibition induced by rostral pons stimulation occurred 3.4 ms after the onset of single electrical pulse stimulation. Based on these results a neuronal model for a pontine mechanism terminating inspiration is proposed.     

Lateralized phrenic nerve responses to stimulating respiratory afferents in the cat.

We stimulated electrically pharyngeal branch of both glossopharyngeal nerves (PGLN), internal branch of superior laryngeal nerves (ISLN), and carotid sinus nerves (CSN) in anesthetized cats. We recorded simultaneously, averaged, and compared bilaterally evoked phrenic nerve (PHR) activity. Our objective was to demonstrate a short-latency evoked response in the PHR contralateral to the stimulus. Low-intensity stimulation of PGLN and ISLN during inspiration evoked a short-latency contralateral excitation with a latency of 5.2 ms +/- 0.2 SE (16 cats) for PGLN, and 3.8 ms +/- 0.1 SE (13 cats) for ISLN. This excitation could follow stimuli delivered at 100 Hz. Stimulation during expiration did not result in a lateralized excitation. The excitation is followed by bilateral inhibition. Neither strychnine nor picrotoxin prevented either the lateralized response or the inhibition, though strychnine diminished a delayed bilateral excitation following PGLN stimulation. This dalayed (latency 18.7 ms +/- 0.7 SE) bilateral excitation corresponds to the sniff reflex. CSN stimulation did not result in lateralized excitation. We suggest that the lateralized evoked response results from a gated paucisynaptic reflex pathway involving the PGLN and ISLN, ipsilateral inspiratory neurons, and contralateral PHR motoneurons.     

Afferent and efferent connections of cat omnipause neurons

Summary Afferent and efferent connections of behaviorally identified omnipause neurons involved in saccadic eye movements were investigated electrophysiologically in cats anesthetized with ketamine hydrochloride. Pause cells were polysynaptically excited by electrical stimulation of the optic chiasm (mean latency = 8.3 ms), the visual cortex (mean latency = 7.3 ms), and the superior colliculus (mean latency = 2.6 ms). Bilateral removal of either the visual cortex or the superior colliculus 1 week prior to the experiment abolished optic chiasm responses. Pause cells were antidromically activated by electrical stimulation of the prerubral fields (mean latency = 1.1 ms), or the pontine and medullary reticular formation (mean latency = 1.0 ms). Frequently, the same pause cell was antidromically excited by prerubral and pontine or medullary reticular stimulation indicating that its axon was branched. The spontaneous discharge of pause cells was polysynaptically suppressed by sustained galvanic polarization of either labyrinth, or by multiple shock stimulation in the reticular formation.     

Spinal connections of ventral-group bulbospinal inspiratory neurons studied with cross-correlation in the decerebrate rat

We examined the synaptic connections from ventral-group bulbospinal inspiratory neurons to upper cervical inspiratory neurons and phrenic and intercostal motoneurons in decerebrate rats using cross-correlation. Inspiratory neurons were recorded in the medulla (n=28) at the level of the obex and from the upper-cervical segments (C1 and C2) of the spinal cord (n=29) in 18 vagotomized, paralyzed, ventilated, and decerebrated rats. The neurons were identified by their inspiratory firing pattern and antidromic activation from the spinal cord at C7. Whole-nerve recordings were made using bipolar electrodes from the central cut ends of the C5 phrenic nerve and the external and internal intercostal nerves at various thoracic levels. Cross-correlation histograms were computed between these recordings to detect short time scale synchronizations indicative of synaptic connections. Cross-correlation histograms (n=20), computed between the activities of ventral-group bulbospinal inspiratory neurons and the phrenic nerve, all showed peaks (mean half-amplitude width±SD, 1.1±0.3 ms) at short latencies (mean latency±SD, 2.0±0.6 ms) suggestive of monosynaptic excitation. Cross-correlation histograms (n=33), computed between the activities of ventral-group bulbospinal inspiratory neurons and upper-cervical inspiratory neurons, displayed four (12%) peaks (mean halfamplitude width±SD, 0.9±0.1 ms) at short latencies(mean latency±SD, 1.8±0.6 ms) suggestive of monosynaptic excitation, and six (18%) peaks (mean half-amplitude width±SD, 1.4±0.4 ms) at latencies near zero suggestive of excitation fro m a common source. Cross-correlation histograms (n=34), computed between the activities of ventral-group bulbospinal inspiratory neurons and the internal and external intercostal nerves at various thoracic levels (T2-8), showed six (18%) peaks (mean half-amplitude width±SD, 2.5±0.5 ms) at short latency (mean latency±SD, 4.5±1.1 ms) suggestive of oligosynaptic connections. Cross-correlation histograms (n=42) computed between activities of intercostal nerves at various levels of the thoracic spinal cord showed central peaks suggestive of excitation from a common source. Although the size of the peaks decreased with segmental separation, the displacement of the peaks from time zero did not increase with segmental separation (mean displacement±SD, 0.6±0.6 ms) as would be expected if the common excitation resulted from a descending monosynaptic excitation by a source such as the ventral-groupbulbospinal inspiratory neurons. We conclude that all ventral-group bulbospinal inspiratory neurons make monosynaptic connections to phrenic motoneurons, a few make monosynaptic connections to upper-cervical inspiratory neurons, but connections to intercostal motoneurons are made via interneurons.     

Inspiratory on-switch evoked by stimulation of the mesencephalon: Activity of phrenic and laryngeal motoneurones

Summary In anaesthetized cats (chloralose-urethan) the effects of brief tetanic electrical stimulation (50 to 100 ms) of the mesencephalic central gray matter and reticular formation on the inspiratory on-switch were studied during the expiratory (E) phase on the gross and unitary activities of phrenic, laryngeal inspiratory and laryngeal expiratory nerves. On the inspiratory laryngeal and phrenic nerves, stimulation elicited a short latency gross response concomitant with the train: the inspiratory Primary Response (Prim.R.) which is followed by an inspiratory Patterned Response (Patt.R.) of longer duration which corresponded to the inspiratory on-switch. The Patt.R. generally appeared from the Prim.R. within a latent period (Silent Phase: Sil.P.) as long as 100 ms. On the expiratory laryngeal nerve, stimulation elicited a brief activation (expiratory Prim.R.) concomitant with the beginning of the inspiratory laryngeal Prim.R. and which rapidly stopped as the latter continued during the stimulus train. The inspiratory Prim.R. corresponded to a simultaneous activation of both early and late (so defined during their spontaneous discharge) inspiratory motoneurones. The laryngeal motoneurones were more strongly activated than the phrenic ones. During the inspiratory Patt.R. all the phrenic motoneurones presented a recruitment delay earlier, compared with the spontaneous one, whereas the recruitment drastically changed from an inspiratory laryngeal motoneurone to another. Thus, the two pools of motoneurones presented different properties of activation. During the inspiratory Sil.P. no concomitant expiratory laryngeal activation was observed when most of the inspiratory motoneurones were inactive. As some inspiratory laryngeal motoneurones did not stop firing, the existence of some central respiratory neurones exhibiting a similar persistent activity and subserving the inspiratory on-switch mechanisms may be hypothesized.Supported by CNRS (LA 205 and ATP n 4188) and Fondation pour la Recherche Médicale     

Role of the ventrolateral region of the nucleus of the tractus solitarius in processing respiratory afferent input from vagus and superior laryngeal nerves

Summary The role of respiratory neurons located within and adjacent to the region of the ventrolateral nucleus of the tractus solitarius (vlNTS) in processing respiratory related afferent input from the vagus and superior laryngeal nerves was examined. Responses in phrenic neural discharge to electrical stimulation of the cervical vagus or superior laryngeal nerve afferents were determined before and after lesioning the vlNTS region. Studies were conducted on anesthetized, vagotomized, paralyzed and artificially ventilated cats. Arrays of 2 to 4 tungsten microelectrodes were used to record neuronal activity and for lesioning. Constant current lesions were made in the vlNTS region where respiratory neuronal discharges were recorded. The region of the vlNTS was probed with the microelectrodes and lesions made until no further respiratory related neuronal discharge could be recorded. The size and placement of lesions was determined in subsequent microscopic examination of 50 m thick sections. Prior to making lesions, electrical stimulation of the superior laryngeal nerve (4–100 A, 10 Hz, 0.1 ms pulse duration) elicited a short latency increase in discharge of phrenic motoneurons, primarily contralateral to the stimulated nerve. This was followed by a bilateral decrease in phrenic nerve discharge and, at higher currents, a longer latency increase in discharge. Stimulation of the vagus nerve at intensities chosen to selectively activate pulmonary stretch receptor afferent fibers produced a stimulus (current) dependent shortening of inspiratory duration. Responses were compared between measurements made immediately before and immediately after each lesion so that changes in response efficacy due to lesions per se could be distinguished from other factors, such as slight changes in the level of anesthesia over the several hours necessary in some cases to complete the lesions. Neither uni- nor bi-lateral lesions altered the efficacy with which stimulation of the vagus nerve shortened inspiratory duration. The short latency excitation of the phrenic motoneurons due to stimulation of the superior laryngeal nerve was severely attenuated by unilateral lesions of the vlNTS region ipsilateral to the stimulated nerve. Neither the bilateral inhibition nor the longer latency excitation due to superior laryngeal nerve stimulation was reduced by uni- or bi-lateral lesions of the vlNTS region. These results demonstrate that extensive destruction of the region of the vlNTS: a) does not markedly affect the inspiratory terminating reflex associated with electrical stimulation of the vagus nerve in a current range selective for activation of pulmonary stretch receptor afferents, and b) abolishes the short-latency increase, but not the bilateral decrease or longer latency increase in phrenic motoneuronal discharge which follows stimulation of the superior laryngeal nerve. We conclude that respiratory neurons in the region of the vlNTS do not play an obligatory role in the respiratory phase transitions in this experimental preparation. Neurons in the vlNTS region may participate in other reflexes, such as the generation of augmented phrenic motoneuronal discharge in response to activation of certain superior laryngeal or vagus nerve afferents.     

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.     

Dopamine1 receptor agonists reverse opioid respiratory network depression, increase CO2 reactivity

In adult pentobarbital-anesthetized and unanesthetized decerebrate cats, the D(1)R agonists (6-chloro-APB, SKF-38393, dihydrexidine) given intravenously restored phrenic nerve and vagus nerve respiratory discharges and firing of bulbar post-inspiratory neurons after the discharges were abolished by the micro-opioid receptor agonist fentanyl given intravenously. Reversal of opioid-mediated discharge depression was prevented by the D(1)R antagonist SCH23390. Iontophoresis of the micro-opioid receptor agonist DAMGO depressed firing of medullary bulbospinal inspiratory neurons. Co-iontophoresis of SKF-38393 did not restore firing and had no effect on bulbospinal inspiratory neuron discharges when applied alone. The D(1)R agonists given intravenously prolonged and intensified phrenic nerve and bulbospinal inspiratory neuron discharges. They also increased reactivity to CO(2) by lowering the phrenic nerve apnea threshold and shifting the phrenic nerve-CO(2) response curve to lower et(CO(2)) levels. Intravenous fentanyl on the other hand decreased CO(2) reactivity by shifting the phrenic nerve apnea threshold and the response curve to higher et(CO(2)) levels. Fentanyl effects on reactivity were partially reversed by D(1)R agonists.     

Nasal trigeminal inputs release the A5 inhibition received by the respiratory rhythm generator of the mouse neonate

Experiments were performed on neonatal mice to analyze why, in vitro, the respiratory rhythm generator (RRG) was silent and how it could be activated. We demonstrated that in vitro the RRG in intact brain stems is silenced by a powerful inhibition arising from the pontine A5 neurons through medullary alpha(2) adrenoceptors and that in vivo nasal trigeminal inputs facilitate the RRG as nasal continuous positive airway pressure increases the breathing frequency, whereas nasal occlusion and nasal afferent anesthesia depress it. Because nasal trigeminal afferents project to the A5 nuclei, we applied single trains of negative electric shocks to the trigeminal nerve in inactive ponto-medullary preparations. They induced rhythmic phrenic bursts during the stimulation and for 2-3 min afterward, whereas repetitive trains produced on-going rhythmic activity up to the end of the experiments. Electrolytic lesion or pharmacological inactivation of the ipsilateral A5 neurons altered both the phrenic burst frequency and occurrence after the stimulation. Extracellular unitary recordings and trans-neuronal tracing experiments with the rabies virus show that the medullary lateral reticular area contains respiratory-modulated neurons, not necessary for respiratory rhythmogenesis, but that may provide an excitatory pathway from the trigeminal inputs to the RRG as their electrolytic lesion suppresses any phrenic activity induced by the trigeminal nerve stimulation. The results lead to the hypothesis that the trigeminal afferents in the mouse neonate involve at least two pathways to activate the RRG, one that may act through the medullary lateral reticular area and one that releases the A5 inhibition received by the RRG.     

Identification of the medullary swallowing regions in the rat

Summary The aim of the present study was to identify the central structures involved in the organization of the swallowing reflex in the rat. Using concentric bipolar electrodes, the medulla and pons were systematically explored in order to determine which central areas responded to stimulation by inducing swallowing. These areas, which were located in the dorsal medulla oblongata, were the solitary tract, the nucleus of the solitary tract (NST) and the adjacent reticular formation. Stimulation of the ventral ponto-medullary regions was ineffective with regard to the initiation of the swallowing reflex. The activity of medullary swallowing neurons was recorded using extracellular microelectrodes. These swallowing neurons responded with a burst of spikes (swallowing activity) which was closely linked to the swallowing reflex elicited by stimulation of the superior laryngeal nerve (SLN). Under SLN stimulation, the activity of some of the swallowing neurons furthermore showed an initial response consisting of 1 or 2 spikes after a brief latency. According to their location and the latency of their initial response, swallowing neurons were divided into two groups. Group I neurons were located in a dorsal area of the medulla oblongata corresponding to the NST and the adjacent reticular formation. All these neurons exhibited an initial response with a very short latency (1 to 4 ms), the swallowing activity of most of these neurons started before the onset of the swallowing motor sequence. Group II neurons were located either in a ventral area corresponding to the nucleus ambiguus and the surrounding reticular formation or in a dorsal and medial area corresponding to the hypoglossal nucleus and its vicinity. Some of these neurons also exhibited an initial response to SLN stimulation, but with a longer latency (7–12 ms). Motor paralysis of the animal (performed by curare injection) did not affect the swallowing activity of the neurons belonging to either group. Thus, the swallowing activity of the medullary neurons studied was a truly central activity. It is concluded that the swallowing neurons studied belonged to the medullary swallowing center; the group II neurons were motoneurons and interneurons forming the efferent stage of the swallowing center, and the group I neurons were the interneurons which largely belong to the center which programs the swallowing motor sequence.This study was supported, in part, by grants from CNRS (LA 205) and INRA     

Short latency inputs to phrenic motoneurones from the sensorimotor cortex in the cat

Summary Short latency responses were recorded from C₅ phrenic roots and intracellularly from phrenic motoneurones following stimulation of the pericruciate cortex or medullary pyramids in cats anaesthetized with Nembutal or chloralose-urethane. Focal stimulation of the cortical surface (single pulses, 0.5–2 ms, 0.3–8 mA) during inspiration evoked EPSPs (latency 4.7 ± 1.7 ms, rise time 1.9 ± 1.1 ms, amplitude 0.22 to 3.94 mV) in 42% of motoneurones studied (n = 107). The EPSPs were absent, or on average 60% smaller, following stimulation during expiration. In all but two motoneurones, during both inspiration and expiration, hyperpolarizing potentials were observed either following the initial depolarization or alone. They could be reversed by hyperpolarizing current or chloride injection. Stimulation of the pyramidal tract at mid medullary level (1 to 3 pulses, 0.2 ms) evoked short latency excitation in phrenic motoneurones only with currents of more than 200 A. Smaller stimuli applied to the medial reticular formation above the pyramidal tract evoked excitation (onset latency 1.5–3.2 ms) in which the earliest part was probably monosynaptic. These results show that the corticospinal responses in phrenic motoneurones are both excitatory and inhibitory. They are not transmitted through the pyramidal tract and are at least disynaptic. Excitation evoked from the medullary pyramidal tract can be explained by current spread beyond the pyramidal tract fibres.     

The role of dorsal respiratory group neurons studied with cross-correlation in the decerebrate rat

We examined the role of dorsal respiratory group (DRG) inspiratory neurons as transmitters of respiratory drive to phrenic and intercostal motoneurons and as relays of afferent information to ventral respiratory group (VRG) bulbospinal, inspiratory neurons. Attempts to antidromically activate 76 DRG neurons from the spinal cord at the C7 segment resulted in only 4 (5.3%) successes (3 contralateral, 1 ipsilateral). Cross-correlating DRG neuron discharge with that of the ipsilateral (56) and contralateral (20) phrenic nerve detected common activation peaks in 2 and 3 cases respectively, with no evidence for monosynaptic connections. Cross-correlating DRG neuron discharge with that of bulbospinal, inspiratory VRG neurons found some evidence for interaction. Peaks in 7 of 73 (10%) cross-correlation histograms were attributed to a monosynaptic excitation of DRG neurons by VRG neurons, although a common activation cannot be ruled out; troughs, some with an accompanying peak, in 9 (12.3%) histograms were interpreted as a combined excitation of the DRG neuron and delayed inhibition of the VRG neuron. In addition, 2 cross-correlation histograms showed peaks with latencies and half-amplitude widths consistent with a disynaptic excitation of a DRG neuron by a bulbospinal inspiratory VRG neuron. Cross-correlating the discharge of 57 pairs of DRG inspiratory neurons (6 contralateral) detected common activation peaks in 7 (12.3%) cases (none contralateral) and one case interpreted as evidence for a disynaptic excitation. These findings suggest that the role of the DRG inspiratory neurons in rats differs from that in cats, primarily because they do not act to transmit respiratory rhythmic drive directly to phrenic and intercostal motoneurons. The results offer some support for an excitation of DRG neurons by VRG inspiratory neurons, but no support for a role of DRG inspiratory neurons as mediators of afferent information transfer to VRG bulbospinal inspiratory neurons.     

Direct and synaptic responses of the cat's medullary respiratory neurons evoked by microstimulation

The responses of single medullary neurons were evoked by microstimulation and recorded extracellularly in cats with spontaneous breathing under pentobarbital anaesthesia. Direct and synaptic responses were observed. In the silent phases of the respiratory neurons the latencies of direct responses were always longer (0.25–0.5 ms) than during the spontaneous discharge of these neurons (0.12–0.35 ms). Such phase-related variations of latency persist even when maximal currents are applied.The latencies of synaptic responses of expiratory and inspiratory neurons differed. The expiratory neurons gave two types of synaptic responses with a latency of about 0.8 ms (monosynaptic) or about 1.5 ms (probably disynaptic). The latencies of the evoked monosynaptic responses of the expiratory neurons varied from 0.6 to 1.2 ms depending on the phase of the respiratory cycle. The minimal latency of the synaptic responses of the inspiratory neurons under study was 1.2 ms. The typical latency varied between 1.75 and 2.8 ms. Some neurons had the late synaptic responses with a latency of about 5–9 ms. The synaptic responses of all the inspiratory neurons as well as the disynaptic responses of the expiratory neurons were evoked only during a spontaneous discharge which was followed by the inhibition of the spontaneous activity (lasting 20–100 ms).Thus it can be concluded that the excitatory pathways to expiratory neurons activated by microstimulation are mono- and disynaptic, whereas excitatory pathways to inspiratory neurons are polysynaptic.     

Responses of feline medial medullary reticulospinal neurons to cardiac input

1. Responses of medial medullary reticulospinal (RS) neurons to electrical stimulation of cardiac sympathetic afferents, probing the epicardium and epicardial application of bradykinin, were determined in cats anesthetized with alpha-chloralose. Conduction velocity of RS cells averaged 67 m/s; these neurons were probably part of the RS motor pathway and the bulbospinal pathway that modulates ascending information. Fifty-three RS neurons had unilateral projections and 18 neurons bilateral projections to the thoracic spinal cord. 2. Maximal electrical stimulation of the left stellate ganglion excited 69% of 97 RS neurons with a mean latency of 15 +/- 1.0 ms. Mean spike discharge rate increased from 4 +/- 1.2 to 93 +/- 8.0 spikes/s for responsive neurons. 3. Epicardial bradykinin (0.04 mg) excited 34%, inhibited 2%, and did not affect 64% of 44 RS cells tested. Excited neurons increased their mean discharge rate from 12.3 +/- 3.6 to 18.2 +/- 4.3 spikes/s, with a latency of 24 +/- 3.0 s, in response to bradykinin. Response duration averaged 43 +/- 2.3 s. Neurons responsive to bradykinin had greater spontaneous discharge rates than unresponsive neurons. Thirteen of 16 RS cells excited by bradykinin were located in the gigantocellular tegmental field (FTG); the other three cells were in the paramedian nucleus (PR). 4. Epicardial bradykinin often elicited changes in aortic pressure. Although some neurons responded to altered blood pressure alone, these responses could not account for responses to bradykinin. Furthermore, the percentage of responsive neurons was similar in experiments with intact nerves as well as those with vagotomy and barodenervation. 5. Touching the epicardium with a blunt probe excited 19 of 60 (32%) RS neurons, and visceral receptive fields were mapped for 12 of these cells. Neurons responded with one to three spikes to probing. RS neurons responsive to probing were scattered throughout the medial reticular formation. 6. RS cells were also tested for somatic, auditory, and visual input. Of 63 neurons responsive to sympathetic afferent stimulation, only one did not receive convergent input from at least one of these sources; 51% received input from each tested source. Neurons responsive to bradykinin were more likely to receive visual input than the general population of neurons. RS neurons unresponsive to sympathetic afferent stimulation were less likely to receive convergent inputs from other sensory modalities.(ABSTRACT TRUNCATED AT 400 WORDS)     

Atonia-related regions in the rodent pons and medulla

Electrical stimulation of circumscribed areas of the pontine and medullary reticular formation inhibits muscle tone in cats. In this report, we present an analysis of the anatomical distribution of atonia-inducing stimulation sites in the brain stem of the rat. Muscle atonia could be elicited by electrical stimulation of the nuclei reticularis pontis oralis and caudalis in the pons as well as the nuclei gigantocellularis, gigantocellularis alpha, gigantocellularis ventralis, and paragigantocellularis dorsalis in the medulla of decerebrate rats. This inhibitory effect on muscle tone was a function of the intensity and frequency of the electrical stimulation. Average latencies of muscle-tone suppressions elicited by electrical stimulation of the pontine reticular formation were 11.02 +/- 2.54 and 20.49 +/- 3.39 (SD) ms in the neck and in the hindlimb muscles, respectively. Following medullary stimulation, these latencies were 11.29 +/- 2.44 ms in the neck and 18.87 +/- 2. 64 ms in the hindlimb muscles. Microinjection of N-methyl-D-aspartate (NMDA, 7 mM/0.1 microliter) agonists into the pontine and medullary inhibitory sites produced muscle-tone facilitation, whereas quisqualate (10 mM/0.1 microliter) injection induced an inhibition of muscle tone. NMDA-induced muscle tone change had a latency of 31.8 +/- 35.3 s from the pons and 10.5 +/- 0. 7 s from the medulla and a duration of 146.7 +/- 95.2 s from the pons and 55.5 +/- 40.4 s from the medulla. The latency of quisqualate (QU)-induced reduction of neck muscle tone was 30.1 +/- 37.9 s after pontine and 39.5 +/- 21.8 s after medullary injection. The duration of muscle-tone suppression induced by QU injection into the pons and medulla was 111.5 +/- 119.2 and 169.2 +/- 145.3 s. Smaller rats (8 wk old) had a higher percentage of sites producing muscle-tone inhibition than larger rats (16 wk old), indicating an age-related change in the function of brain stem inhibitory systems. The anatomical distribution of atonia-related sites in the rat has both similarities and differences with the distribution found in the cat, which can be explained by the distinct anatomical organization of the brain stem in these two species.     

The differential organization of medullary post-inspiratory activities

Membrane potential trajectories of 68 bulbar respiratory neurones from the peri-solitary and peri-ambigual areas of the brain-stem were recorded in anaesthetized cats to explore the synaptic influences of post-inspiratory neurones upon the medullary inspiratory network.A declining wave of inhibitory postsynaptic potentials resembling the discharge of postpinspiratory neurones was seen in both bulbospinal and non-bulbospinal inspiratory neurones, including alpha- and beta-inspiratory, early-inspiratory, late-inspiratory and ramp-inspiratory neurones.Activation of laryngeal and high-threshold pulmonary receptor afferents excited bulbar post-inspiratory neurones, whilst in the case of inspiratory neurones such stimulation produced enhanced postsynaptic inhibition during the same period of the cycle. Activation of post-inspiratory neurones and enhanced post-inspiratory inhibition of inspiratory bulbospinal neurones was accompanied by supression of the after-discharge of phrenic motoneurones.These results suggest that a population of post-inspiratory neurones exerts a widespread inhibitory function at the lower brain-stem level. Implications of such an inhibitory function for the organization of the respiratory network are discussed in relation to the generation of the respiratory rhythm.     

Hypoglossal premotor neurons with rhythmical inspiratory-related activity in the cat: localization and projection to the phrenic nucleus

Localization and projection to the phrenic (PH) nucleus were studied in a sample of premotor neurons that directly projected to hypoglossal motoneurons (XII Mns) and showed respiratory-related patterns of activity. The experiments were carried out in cats, under pentobarbital anesthesia. In the first part of the study, the retrograde double-labeling technique was used to reveal the existence of neurons projecting to both the XII and the PH nuclei. Injection of a fluorescent dye (fast blue, FB) into the XII nucleus and another (nuclear yellow, NY) into the PH nucleus retrogradely labeled, with either FB or NY, medullary reticular neurons mainly in the regions ventrolateral to the nucleus of the tractus solitarius (vl-NTS), ventrolateral to the hypoglossal nucleus (vl-XII), and dorsomedial to the nucleus ambiguus (dm-AMB) bilaterally. In addition, some neurons in these regions were labeled with both FB and NY. In the second part of the study, unitary activity was recorded extracellularly from medullary respiratory neurons. In the regions vl-NTS, vl-XII, and dm-AMB, inspiratory neurons were found which antidromically responded to stimulation of the XII nucleus. Some of them also responded antidromically to stimulation of the PH nucleus. Averaging of rectified and integrated XII and PH nerve discharges by spontaneous spikes of single inspiratory neurons in the vl-NTS and dm-AMB regions revealed a facilitation in either XII nerve discharge or both XII and PH nerve discharges after a short latency of monosynaptic range. It is concluded that in the vl-NTS and dm-AMB regions there are inspiratory neurons that are excitatory premotor neurons projecting to XII Mns showing the respiratory-related activity. Some of them have excitatory synaptic connections to XII and PH Mns via bifurcating axons.     

The effect of procaine injection into the medullary reticular formation on forelimb muscle activity evoked by mesencephalic locomotor region and vestibular stimulation in the decerebrated guinea-pig.

The effect of procaine microinjection into the ventromedial portion of the medullary reticular formation on forelimb muscle activity evoked by electrical mesencephalic locomotor region and natural vestibular stimulation has been investigated in the decerebrated guinea-pig. This injection is followed by a reversible increase of the threshold of mesencephalic locomotor region stimulation necessary for the activation of muscle rhythmic activity. Procaine injection is accompanied by reduction of vestibular influence on flexor muscle activity evoked by electrical cutaneous and mesencephalic locomotor region stimulation. Vestibular influence on extensor muscle activity remains unchanged after the injection. The results indicate that medial medullary reticular formation is the site of the convergence of mesencephalic locomotor region and vestibular activity. It is suggested that the vestibular system contributes to the modulation of reticulospinal activity coupled with the initiation and control of locomotion.     

Functional distinction of perigeniculate and thalamic reticular neurons in the cat

Summary Two types of neurons can be recognized in the region above the lateral geniculate nucleus. One cell type is found in the caudal part of the reticular nucleus of thalamus; these cells are accordingly called reticular neurons. The other cell type is located in the perigeniculate nucleus immediately above lamina A of the lateral geniculate nucleus and in the intermediate zone between the perigeniculate nucleus and the reticular nucleus. These cells are referred to as perigeniculate neurons. Electrical stimulation of the optic tract and the visual cortex typically evokes a short burst of spikes in the perigeniculate neurons, and the excitation has a shorter latency from the cortex (range 1.2–2.5 ms) than from the optic tract (range 1.5–3.1 ms). The perigeniculate neurons are also activated by adequate visual stimuli. In contrast, the reticular neurons are unresponsive to visual stimuli and electrical stimulation of the optic tract but they may respond with a burst of spikes to cortex stimulation with rather long latency (range 2.7–5.5 ms). It is concluded that only perigeniculate neurons qualify as interneurons in the recurrent inhibitory pathway to principal cells in the lateral geniculate nucleus.Supported by Magnus Bergvalls Stiftelse and The Swedish Medical Research Council (Project no. 4767)F.-S. Lo had an exchange fellowship from the Royal Swedish Academy of Engineering