Analysis of reflex activity in cardiac sympathetic nerve induced by myelinated phrenic nerve afferents

Electrical stimulation of the phrenic nerve afferents evoked excitatory responses in the right inferior cardiac sympathetic nerve in chloralose-anaesthetized cats. The reflex was recorded in intact and spinal cats. The latency and threshold of the volley recorded from the phrenic nerve as well as of the cord dorsum potentials evoked by electrical stimulation of the phrenic nerve indicated that group III afferents were responsible for this reflex. The phrenicocardiac sympathetic reflex recorded in intact cats was followed by a silent period. The maximum amplitude of the reflex discharges was 800 microV, the latency was 83 ms and the central transmission time 53 ms. Duration of the silent period lasted up to 0.83 s. In spinal cats the reflex was recorded 5.5-8 h after spinalization. The maximum amplitude of the spinal reflex discharges ranged from 22 to 91 microV and the latency from 36 to 66 ms.     

Supraspinal involvement in the phrenic-to-phrenic inhibitory reflex

Phrenic afferents are capable of attenuating the phrenic motor response elicited by the intercostal-to-phrenic excitatory reflex in decerebrate, paralyzed cats. High spinal transection eliminates the attenuating effect of the bilateral phrenic-to-phrenic inhibitory reflex. These results indicate that although phrenic nerve afferents do exert an inhibitory influence in the cervical spinal cord, some of the inhibitory effects are likely to involve supraspinal mechanisms.     

Neural elements subserving pulmonary stretch receptor-mediated facilitation of phrenic motoneurons

The neural elements responsible for facilitation of phrenic nerve activity by lung inflation were investigated in cats by the simultaneous recording of individual pulmonary stretch receptor afferents, respiratory neurons of the ventrolateral nucleus of the tractus solitarius and phrenic nerve activity. Monosynaptic excitation of I beta neurons by slowly adapting pulmonary stretch receptors was demonstrated by cross-correlational analysis. It was also demonstrated that the majority of these same I beta neurons projected to the contralateral C5 phrenic motoneuron pool. Thus, this study has shown that I beta neurons can act as central neural elements to mediate the facilitatory effect of lung inflation upon phrenic nerve activity.     

Pulmonary vagal sensory afferents and spontaneous EEG rhythms in the cat sensorimotor cortex

Interactions between vagal afferent fibres and spontaneous electroencephalographic (EEG) activity, recorded on the sensorimotor cortex of the cat, were studied during the mechanical activation of pulmonary afferents. The interactions were compared to the cortical effects of the electrical stimulation of all vagal fibers or to the chemical activation of unmyelinated vagal afferents (C-fibers) by phenyldiguanide. The present study was performed on anesthetized cats, artificially ventilated with open chest. Over 60 locations were explored on the posterior sigmoid gyrus. Repetitive electrical stimulation (30 Hz, 0.8 ms shock duration) of the contralateral cervical vagus nerve or of both nerves induced within less than 5 s changes in the pattern and periodicity of EEG spindles, associated with depressed background rhythms or rhythmic EEG activities. Cortical responses were also observed after i.v. injection of phenyldiguanide. Changes in activity of pulmonary stretch receptors by lung hyperinflation or suppression of phasic lung inflations ('stop pump') had no effect on the EEG rhythms. On the other hand, expiratory threshold loading or passive hyperdeflation of the lungs elicited EEG changes similar to those obtained by electrical stimulation of all vagal fibers. After bilateral vagotomy, all these responses disappeared or were delayed. The present observations strongly suggest that sensory information carried by thin vagal fibers greatly influences cortical rhythms in the cat sensorimotor cortex.     

Restorative respiratory pathways after partial cervical spinal cord injury: role of ipsilateral phrenic afferents

After disruption of the descending respiratory pathways induced by unilateral cervical spinal cord injury (SCI) in rats, the inactivated ipsilateral (ipsi) phrenic nerve (PN) discharge may partially recover following some specific experimental procedures [such as contralateral (contra) phrenicotomy (Phx)]. This phrenic reactivation involves normally silent contra pathways decussating at the level of the phrenic nucleus, but the mechanisms of this crossed phrenic activation are still poorly understood. The present study investigates the contribution of sensory phrenic afferents to this process by comparing the acute effects of ipsi and contra Phx. We show that the phrenic discharge (recorded on intact PNs) was almost completely suppressed 0 h and 3 h after a lateral cervical SCI, but was already spontaneously reactivated after 1 week. This ipsi phrenic activity was enhanced immediately after contra Phx and was completely suppressed by an acute contra cervical section, indicating that it is triggered by crossed phrenic pathways located laterally in the contra spinal cord. Ipsi phrenic activity was also abolished immediately after ipsi Phx that interrupts phrenic sensory afferents, an effect prevented by prior acute ablation of the cervical dorsal root ganglia, indicating that crossed phrenic activation depends on excitatory phrenic sensory afferents but also putatively on inhibitory non-phrenic afferents. On the basis of these data, we propose a new model for crossed phrenic activation after partial cervical injury, with an essential role played by ipsi-activating phrenic sensory afferents.     

Behavior of inhibitory and excitatory propriobulbar respiratory neurons during fictive vomiting.

The behavior of propriobulbar respiratory neurons was studied during fictive vomiting in decerebrate, paralyzed, artificially ventilated cats. Fictive vomiting was identified by a characteristic series of synchronous phrenic and abdominal nerve bursts, induced by electrical stimulation of abdominal vagal afferents and/or i.v. infusion of emetic drugs. Data were obtained from inspiratory neurons having decrementing (I-DEC) or constant (I-CON) discharge patterns and expiratory decrementing (E-DEC) neurons located in the B?tzinger complex and adjacent rostral ventral respiratory group. These neurons are known to make excitatory (I-CON) and inhibitory (I-DEC, E-DEC) connections with a variety of medullary respiratory neurons. During fictive vomiting: 8 of 14 I-DEC neurons fired in phase with synchronous bursts of phrenic and abdominal nerve discharge; the other 6 were silent. Of 12 I-CON neurons, 5 fired in phase with phrenic and abdominal bursts; 7 were silent. All (6) E-DEC neurons were either silent or fired weakly between bursts of phrenic and abdominal discharges. The possible roles of I-DEC and I-CON neurons in actively reorganizing the behavior of other respiratory neurons during fictive vomiting are discussed. In particular, the firing of many I-DEC neurons was found to be appropriate to inhibit inspiratory, and two types of expiratory, bulbospinal neurons during fictive vomiting.     

Development of callosal connections in the sensorimotor cortex of the hamster.

To investigate the development of corpus callosal connectivity in the hamster sensorimotor cortex, we have used the sensitive axonal tracer 1,1 dioctadecyl-3,3,3',3', tetramethylindocarbocyanine perchlorate (DiI), which was injected either in vivo or in fixed brains of animals 3-6 days postnatal. First, to study changes in the overall distribution of developing callosal afferents we made large injections of DiI into the corpus callosal tract. We found that the anterogradely labeled callosal axons formed a patchy distribution in the contralateral sensorimotor cortex, which was similar to the pattern of adult connectivity described in earlier studies of the rodent corpus callosum. This result stands in contrast to previous retrograde studies of developing callosal connectivity which showed that the distribution of callosal neurons early in development is homogeneous and that the mature, patchy distribution arises later, primarily as a result of the retraction of exuberant axons. The initial patchy distribution of callosal axon growth into the sensorimotor cortex described in the present study suggests that exuberant axons destined to be eliminated do not enter the cortex. In addition, small injections of DiI into developing cortex resulted in homotopic patterns of callosal topography in which reciprocal regions of sensorimotor cortex are connected, as has been shown in the adult. Second, to study the radial growth of callosal afferents we followed the extension of individual callosal axons into the developing cortex. We found that callosal axons began to invade the contralateral cortex on about postnatal day 3, with little or no waiting period in the callosal tract. Callosal afferents then advanced steadily through the cortex, never actually invading the cortical plate but extending into layers on the first day that they could be distinguished from the cortical plate. The majority of callosal axons grew radially through the cortex and did not exhibit substantial branching until postnatal day 8, the age when the cortical plate disappears and callosal afferents reach the outer layer of cortex. This mode of radial growth through cortex prior to axon branching could serve to align callosal afferents with their radial or columnar targets before arborizing laterally.     

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.     

Anesthesia affects respiratory and sympathetic nerve activities differentially.

Phrenic and cervical sympathetic nerve responses to hypercapnia were examined before and after anesthesia in twelve midcollicularly decerebrated, vagotomized, glomectomized, paralyzed and ventilated cats. We measured responses of integrated phrenic and cervical sympathetic nerve activities to increases in end-tidal PCO2 (PETCO2) from apneic threshold to approximately 30 torr above threshold. All cats were studied first in the unanesthetized state. Six cats were then restudied after a quarter of a usual dose of chloralose/urethane (10 mg/kg and 62.5 mg/kg, respectively) and then after half the usual dose of chloralose/urethane (20 mg/kg and 125 mg/kg). The other six animals were restudied after quarter of a standard dose of pentobarbital (9 mg/kg), after half the standard dose (18 mg/kg) and then after the full (35 mg/kg) dose. Both anesthetic agents led to significant increases in apneic thresholds for both phrenic and sympathetic nerve activities. These agents also caused dose-dependent decreases in peak, tonic and respiratory-related sympathetic nerve activities. Peak (tidal) phrenic nerve activities, in comparison, were much less affected by the anesthetic agents. CO2 response curves showed that both of these anesthetic agents depressed, at any given level of PETCO2, respiratory-related sympathetic nerve responses more than the responses found in the phrenic nerve. We conclude that the relations between peak, tonic (i.e. between phasic bursts) and respiratory-related sympathetic nerve activities and phrenic nerve activity can be altered by anesthesia.     

Motor recovery after cross-reinnervation of a forelimb nerve by the phrenic nerve in cats

Electromyographic (EMG) activities of forelimb muscles and the diaphragm following cross-reinnervation of the elbow flexor nerve by the phrenic nerve sampled simultaneously with cinematographic recording of forelimb movements, in order to examine whether the respiratory nervous system in adult cats is capable of compensation for movement disorders. The right phrenic and musculocutaneous nerves were cut preparatory to cross-union, the proximal stump of the phrenic nerve being joined to the peripheral stump of the musculocutaneous nerve. Reinnervation of the elbow flexor muscles (m. biceps brachii and m. brachialis) was confirmed in terminal experiments by observing muscular contractions induced by nerve stimulations and/or retrogradely labeled motoneurons in cervical (C5-C7) segments following treatment for horseradish peroxidase (HRP) which had been injected previously into the m. biceps brachii of both sides. Behavioral observations were made in the chronically cross-reinnervated cats. Bipolar EMG recording electrodes were implanted chronically in the right m. biceps brachii, m. triceps brachii and in the left intact diaphragm. In freely moving cats 3-22 months after cross-union surgery, EMG activities in the elbow flexor usually were synchronized with those of the left diaphragm. However, occasionally, in cats cross-reinnervated more than 6 months previously, EMG activities in the right biceps muscle were dissociated from those of the left diaphragm during voluntary movements. During walking, EMG activities in the cross-reinnervated biceps muscle were not coordinated with the walking cycle. By contrast, during voluntary goal directed movements, the cats were capable of reaching their right forepaws to the target in a smooth manner using elbow flexion that was associated with EMG activity of the biceps muscles.(ABSTRACT TRUNCATED AT 250 WORDS)     

Consequences of damage to the sensorimotor cortex in neonatal and adult cats. II. Maintenance of exuberant projections

After chronic sensorimotor cortex ablations, sparing and greater recovery of function were seen in neonatally operated cats compared with adult operated cats. These results suggested that undamaged cortex in neonatal operates might display projections different from those of adult operates. Injections of horseradish peroxidase-wheat germ agglutinin (HRP-WGA) were made in ipsilateral parietal cortex adjacent to the sensorimotor cortex ablations or in the contralateral sensorimotor cortex. No changes in the projections of the parietal cortex were seen in operated cats or in the projections of the undamaged sensorimotor cortical projections of adult operates. In contrast, the intact sensorimotor cortex of neonatal operates exhibited crossed corticothalamic and corticorubral projections not present in normal or adult operated animals, whereas the corticospinal tract (CST) was unchanged by the ablations. Analysis of neurons within the ventroanterior-ventrolateral nuclear complex of the thalamus ipsilateral to the ablation showed that the surviving cells of neonatal operates were equal in number but were, on average, larger than those of normals and adult operates. Some neurons in neonatal operates were larger than any seen in adult operates and normals. Injections of HRP/WGA were also made into the sensorimotor cortex of normal newborn animals. Dense bilateral corticothalamic and corticorubral projections were present. The CST had extended to lumbar levels by the day of birth but projections to the grey matter were sparse. Thus, bilateral projections seen in neonatal operates probably represent retention of some exuberant projections present in normal neonatal animals. The CST which exhibited no exuberant projection was unchanged by the lesion. The crossed corticothalamic and corticorubral projections are likely to play a role in sparing and recovery of function particularly in sparing of contact placing.     

Substantia nigra unit responses to trigeminal sensory stimulation.

The influence of trigeminal sensory stimulation on substantia nigra units was investigated in cats anesthetized with chloralose. Low-intensity electrical stimulation was applied to primary sensory afferents (inferior dental nerve, trigeminal mesencephalic nucleus), and a high proportion of substantia nigra units showed evoked changes in activity. Responses were typically excitatory. Peripheral receptive fields were assessed by applying natural mechanosensory and also electrical stimulation to the face, perioral, and intraoral tissue. Substantia nigra units responded to light tactile stimuli; the majority of the receptive fields were bilateral, and all subsumed at least two branches of the trigeminal nerve. Latency considerations suggest that these sensory responses were not conveyed via striatonigral connections. The findings have implications in relation to the role of the basal ganglia, in general, in oropharyngeal sensorimotor processes.     

Involvement of N-methyl-D-aspartate (NMDA) receptors in respiratory rhythmogenesis

The involvement of N-methyl-D-aspartate (NMDA) subtype of glutamate receptors in the control of inspiratory termination was studied in paralyzed decerebrated cats. Cats were either vagotomized, or had intact vagus nerves and were ventilated with a ventilator driven by the discharge of the phrenic nerve. The systemic administration of NMDA antagonists acting non-competitively (MK-801, ketamine, phencyclidine) or competitively (2-amino-7-phosphonoheptanoic acid: AP7), produced an apneusis in vagotomized animals or in animals transiently deprived of vagal pulmonary feedback by the 'no inflation test'. After NMDA receptor blockade, the inspiratory phase could be terminated by lung inflation or sensory stimulation. Thus pharmacologically distinct mechanisms control the termination of inspiration: vagal afferents which are NMDA-independent, and a central mechanism acting through the activation of NMDA receptors. The apneustic pattern induced by NMDA receptor blockade was characterized by a decrease of the amplitude of integrated phrenic nerve activity, the persistence of CO2 sensitivity and an enhancement of apneusis by anaesthesia. After injection of NMDA antagonists there was a decrease of the duration of expiration which thereafter remained constant and dissociated from inspiratory duration. The possible mechanisms by which NMDA receptors may contribute to respiratory rhythmogenesis are discussed.     

Phrenic nerve palsy as a feature of chronic inflammatory demyelinating polyradiculoneuropathy

We report four patients with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) who presented with phrenic nerve palsy, which was unilateral in two instances. The two patients with bilateral phrenic nerve involvement required mechanical ventilation. After treatment with intravenous immunoglobulins (i.v.Ig) or steroids, the sensorimotor deficit and respiratory parameters improved in three patients, but the fourth patient remained ventilator dependent and died from pulmonary infection. Although rare, phrenic nerve palsy may be a feature of CIDP and may be responsive to treatment with i.v.Ig or steroids.     

Role of nucleus retroambigualis in respiratory reflexes evoked by superior laryngeal and vestibular nerve afferents and in emesis

An ascending projection from the medullary nucleus retroambigualis (NRA) has recently been described as important for the control of the upper airway during vocalization. We evaluated the importance of this projection in other behaviors by making localized injections of the neurotoxin kainic acid in the NRA in decerebrate cats, most of which were paralyzed and artificially ventilated. In contrast to its importance for vocalization, the NRA is not essential for activation of upper airway musculature during respiration, swallowing, vomiting, or reflexes elicited by superior laryngeal or vestibular nerve afferents. However, kainic acid injections in the NRA and adjacent reticular formation prolonged the inhibitory phrenic motoneuronal response to superior laryngeal nerve stimulation and abolished or reduced abdominal motoneuronal responses during respiration, vomiting, and superior laryngeal nerve stimulation. Thus, of the behaviors we investigated, the importance of the ascending projection from the NRA appears to be limited to vocalization, while descending projections from the NRA region are important in a number of behaviors.     

Effect of electric and chemical stimulation of the raphe obscurus on phrenic nerve activity in the cat

The effect of electrical and chemical (l-glutamate) stimulation of the raphe obscurus on phrenic nerve activity was examined in the cat. Phrenic nerve activity was recorded from a C5 nerve root in anesthetized, paralyzed and artificially ventilated cats. Neural discharge was quantitated by integrating the phrenic nerve activity. The respiratory frequency was determined from the integrated nerve signal. Focal electrical stimulation (18–144 μA; 5–40 Hz; 100 μs pulse duration) resulted in significant (P < 0.05) increases in both integrated phrenic nerve (IPN) amplitude and respiratory frequency. These changes were dependent upon current intensity and frequency of stimulation. The largest increases in IPN amplitude and respiratory frequency were47 ± 17%and146 ± 8%, respectively. To insure that the changes in integrated phrenic nerve activity (IPNA) were the result of stimulation of cell bodies and not axons of passage,l-glutamate (100, 200 nmol) was microinjected (100 nl) into the raphe obscurus. Significant (P < 0.05) dose-related changes occurred in integrated phrenic nerve amplitude with an increase of44 ± 13% at 100 nmol and80 ± 13% at 200 nmoll-glutamate. No significant increase in respiratory frequency was observed withl-glutamate microinjection. The results suggest that the raphe obscurus may be involved in respiratory control.     

Quantitative EEG changes under various conditions of hyperventilation in the sensorimotor cortex of the anaesthetized cat.

The effects on the EEG rhythms recorded from the sensorimotor cortex (post-sigmoid gyrus) of anaesthetized cats were studied under 4 conditions of artificial mechanical hyperventilation (HV) before and after cervical bilateral vagotomy. In animals with intact vagus nerves, using visual examination, EEG changes were only observed within the 2nd min during HV produced by increased stroke volume (delta V) with associated hypocapnia. Quantitative EEG (qEEG) showed that, for the same increase in minute ventilation and the same degree of hypocapnia, delta V induced a greater and earlier relative decrease (2nd min) in the power density of delta, theta and alpha bands, than increased pump frequency (delta F). The delta F tests produced a fall only in the theta band and within the 3rd min. With constant paCO2, transient modifications occurred only with delta V and were limited to the first 30 sec. In bivagotomized cats, moderate EEG responses to delta V plus associated hypocapnia persisted partly in the alpha band. Finally, no changes appeared with delta V or delta F when the vagus nerves were cut and paCO2 was maintained constant. The present data suggest strongly that, in anaesthetized cats, peripheral vagal afferents from the respiratory system play a major role in the EEG changes caused by artificial hyperventilation.     

Afferent innervation of the gallbladder in the cat, studied by the horseradish peroxidase method

Following injection of horseradish peroxidase (HRP) into the wall of the gallbladder of cats, HRP-positive cells were found bilaterally in dorsal root ganglia T2-L3 (T2-L2, and T3-L2/L3 also observed in a few cats) and nodose ganglia. In about 33% of animals labelled cells were also distributed in cervical dorsal root ganglia C5-C7. Labelled cells were more frequently localized on the right side than the left. There was no apparent change in numbers of labelled cells in the nodose ganglion (NG) on either side following greater and lesser splanchnicotomy or section of the right phrenic nerve or removal of the celiac ganglion. After severing both the greater and lesser splanchnic nerves unilaterally, numbers of labelled afferent cells from the gallbladder in dorsal root ganglia (DRGs) significantly decreased on the ipsilateral side but there was no change in the pattern of distribution contralaterally. After section of the right phrenic nerve, labelled cells were not found in ipsilateral cervical ganglia. That some afferent fibers from the gallbladder travel via the phrenic nerves, particularly on the right side, may be a supplementary mechanism in the generation of referred pain in gallbladder disease. The splanchnic nerves are the main, but not the only pathway for afferent fibers from the gallbladder.     

Efferent activity in the phrenic nerve during the startle reflex in chloralose-anesthetized cats

Reflex activity in the phrenic nerve was studied in chloralose anesthetized cats during development of somatic startle reflexes in limb and lower intercostal nerves. It was shown that the main component of this activity during low-threshold reflexes evoked by acoustic, tactile and low-threshold somatic afferent stimulation was depression of phrenic inspiratory activity. The following reflex discharges were prevalent components of phrenic responses to high-threshold afferent stimulation: early, propriospinal (intercostal-to-phrenic reflex) and late, suprasegmental ones. The latter were of two types: inspiratory (observed mainly during inspiration in about 75% of experiments) and expiratory (observed during expiration in 25% of experiments) which could be classified as "phrenic startle reflexes". Modulation of all responses during the respiratory cycle was described. Structural characteristics of reflex responses evoked in the phrenic nerve by stimulation of various respiratory and nonrespiratory bulbar sites as well as their respiratory modulation have been analyzed. Organization of possible neurophysiological mechanisms of phrenic responses during startle reflexes is discussed.     

Serotonin immunoreactive boutons make synapses with feline phrenic motoneurons

In anesthetized cats, phrenic motoneurons were intracellularly labeled with HRP. Immunohistochemistry was used to localize serotonin-like immunoreactivity that was present in numerous boutons and nerve fibers within the ventral horn of the C5 spinal segment. Immunoreactive boutons were frequently found in apposition to phrenic motoneurons, but these close contacts were more common on the dendrites than the cell body. At the electron microscope level, serotonin-immunoreactive boutons were found to make synapses with well-defined postsynaptic densities on proximal and distal dendrites of phrenic motoneurons. These results suggest that serotonin-containing neurons may directly affect the excitability of phrenic motoneurons, mainly through an input onto their extensive dendritic trees.