Rostral Ventral Respiratory Group Research Articles

Identification of the axon pathways which mediate functional recovery of a paralyzed hemidiaphragm following spinal cord hemisection in the adult rat

Despite extensive neurophysiological work carried out to characterize the crossed phrenic phenomenon, relatively little is known about the morphological substrate of this reflex which restores function to a hemidiaphragm paralyzed by spinal cord injury. In the present study WGA-HRP was injected into normal and functionally recovered hemidiaphragm muscle in rats during the crossed phrenic phenomenon. The retrograde transynaptic transport characteristics of WGA-HRP was utilized to delineate the source of the neurons which mediate the crossed phrenic phenomenon. The results indicated that the neurons which drive phrenic motoneurons in spinal hemisected rats during the crossed phrenic phenomenon are located bilaterally in the rostral ventral respiratory group (rVRG) of the medulla. No transneuronal labeling of propriospinal neurons was noted in either normal or spinal-hemisected rats. Thus, propriospinal neurons do not relay respiratory drive to phrenic motoneurons. The neurons of the rVRG project monosynaptically to phrenic motoneurons. The present results suggest that both crossed and uncrossed bulbospinal pathways from the rVRG collateralize to both the left and right phrenic nucleic and functional recovery of a hemidiaphragm paralyzed by ipsilateral spinal cord hemisection is mediated by supraspinal neurons from both sides of the brain stem. These results are important to our complete understanding of the mechanisms which govern motor recovery in mammals following spinal cord injury.

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 characteristics series of synchoronous 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 Bo¨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 synchornous 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.

Reciprocal connections between rostral ventrolateral medulla and inspiration-related medullary areas in the cat

We investigated connections between the rostral ventrolateral medulla (rVLM) and the two main inspiration-related medullary areas, i.e., the dorsal respiratory group (DRG) and the rostral ventral respiratory group (rVRG) in the cat. Non respiration-related tonically firing units encountered in the rVLM displayed either antidromic or orthodromic responses to DRG or rVRG microstimulation. Some units responded to the stimulation of both regions. We suggest that at least part of rVLM neurons are components of medullary loops operating in the control of breathing.

Decussation of bulbospinal respiratory axons at the level of the phrenic nuclei in adult rats: A possible substrate for the crossed phrenic phenomenon

The axonal trajectories of inspiratory bulbospinal neurons were examined after deposition of the anterograde neuronal tracer phaseolus vulgaris leucoagglutinin (PHA-L) into the rostral ventral respiratory group in rats. At the level of the phrenic nucleus, PHA-L-labeled bulbospinal axons crossed the midline of the spinal cord in both the anterior gray and the anterior white commissure. These spinally decussating neurons provide a possible anatomical substrate for the respiratory reflex known as the crossed phrenic phenomenon.

Subnuclear organization of the lateral tegmental field of the rat. II: Catecholamine neurons and ventral respiratory group

Bulbospinal and propriobulbar respiratory neurons of the ventral respiratory group and catecholamine neurons of the A1 and C1 cell groups were simultaneously labelled in the rat medulla by a combination of retrograde tracing and immunohistochemical identification. The ventral respiratory group and catecholamine cell groups form adjacent, parallel cell columns in the lateral tegmental field of the ventrolateral medulla. The ventral respiratory group is located immediately dorsal to the A1 and C1 groups, although some A1 neurons are intermingled with neurons of the rostral ventral respiratory group, and some C1 neurons are intermingled with those of the Bötzinger complex. The proximate populations of respiratory, catecholamine, and (presumptive) cardiovascular neurons identified in this study provide further support to the hypothesis that this region of the lateral tegmental field of the ventrolateral medulla is a site of cardiorespiratory coordination.

Brainstem connections of the rostral ventral respiratory group of the rat

The pontomedullary connections of the rostral division of the ventral respiratory group (rVRG), the largest medullary population of inspiratory bulbospinal and propriobulbar neurons, were identified in the rat by retrograde and anterograde tracing techniques. These experiments revealed that: (i) the Kölliker-Fuse nucleus, portions of the medial and lateral parabrachial nuclei, and all levels of the ipsilateral and contralateral VRG complex have dense reciprocal connections with the rVRG; (ii) the lateral paragigantocellular nucleus has reciprocal but less dense connections with rVRG; (iii) portions of the nucleus of the solitary tract have prominent projections to, but weaker inputs from rVRG; (iv) the raphe, magnocellular tegmental field and spinal trigeminal nuclei have minor projections to rVRG and receive only sparse inputs from rVRG, and; (v) the retrotrapezoid nucleus, pontine lateral tegmental field and area postrema each have only efferent projections to rVRG. These findings are consistent with previous studies of pontomedullary connections of rVRG in the cat, and further document the extensive reciprocal connections between principal respiratory groups. These connections are likely to be important for generation of the respiratory pattern, for the coordination of effector responses of the cranial and spinal respiratory motor neurons to afferent stimuli, and coordination of the central respiratory and cardiovascular control systems.

Origin of serotonin-containing projections to the ventral respiratory group in the rat

The major purpose of the present study was to determine the origin of the serotonin-containing neurons which project to the rostral ventral respiratory group in the rat. This was accomplished by using the technique of retrograde tracing with rhodamine-labeled latex microspheres (beads) combined with immunohistochemistry. The rhodamine-labeled beads were microinjected into electrophysiologically identified groups of inspiratory neurons in the rostral ventral respiratory group to retrogradely label neurons projecting to this site. Immunohistochemical processing of the tissue was then done to determine if serotonin was present in the retrogradely-labeled neurons. Serotonin-containing neurons projecting to the rostral ventral respiratory group were found in the raphe magnus, raphe obscurus, raphe pallidus and in the paraolivary region extending to the ventral medullary surface. No serotonin-containing neurons in more rostrally located raphe nuclei were found to project to the rostral ventral respiratory group. The findings suggest that caudal raphe serotonergic projections may affect the activity of respiratory neurons in the rostral ventral respiratory group. Projections to the rostral ventral respiratory group from other pontomedullary nuclei were also identified. Rhodamine-labeled neurons were found in the area of the Kölliker-Fuse nucleus, lateral and medial parabrachial nuclei, retrofacial nucleus, nucleus ambiguus/retroambigualis, nucleus tractus solitarius, A5 region, nucleus paragigantocellularis lateralis, retrotrapezoid nucleus, area postrema and spinal trigeminal nucleus. The projections to the rostral ventral respiratory group in the rat are similar to those previously described in the cat and suggest a common circuitry for the CNS control of breathing.

Are there serotonergic projections from raphe and retrotrapezoid nuclei to the ventral respiratory group in the rat?

The relationship of serotonergic neurons in raphe and ventrolateral medullary regions to neurons projecting to the rostral ventral respiratory group in the rat were investigated using combined immunohistochemical and retrograde labeling techniques. Serotonergic and non-serotonergic neurons retrogradely labeled with rhodamine beads were found intermingled in the larger population of serotonin-immunoreactive neurons in raphe pallidus, obscurus and magnus. In addition, a cell group analogous to the retrotrapezoid nucleus in the cat was identified in the rat ventrolateral medulla. Retrotrapezoid neurons, which exhibited exclusively ipsilateral projections to the ventral respiratory group, were located lateral to clusters of serotonergic cells near the ventral surface, but were not serotonin immunoreactive.

Monosynaptic transmission of respiratory drive to phrenic motoneurons from brainstem bulbospinal neurons in rats.

The termination patterns in the rat phrenic nucleus of neurons within two respiratory cell groups of the ventrolateral medulla (Bötzinger Complex and the rostral ventral respiratory group) were determined. The plant lectin, Phaseolus vulgaris leuco-agglutinin, was used as an anterograde tracer to label presynaptic processes of bulbospinal neurons, and horseradish peroxidase was used simultaneously to label phrenic motoneurons. Labeled bulbospinal axons ended with dense terminal arborizations within the phrenic cell column and on radial phrenic motoneuron dendrite bundles, which represented the exclusive site of termination of Bötzinger Complex and rostral ventral respiratory group neurons in the lower cervical spinal cord. Terminals of these descending axons formed presumptive synaptic contacts within longitudinal and radial dendrite bundles, and on the cell somata of phrenic motoneurons.

Locations of medullary neurons with non-phasic discharges excited by stimulation of central and/or peripheral chemoreceptors and by activation of nociceptors in cat

The activity of medullary neurons (146 units) with non-phasic discharges was recorded extracellulary in decerebrated, spontaneously breathing cats. The firing rate changes were studied during injections of 100% CO 2-saturated saline into the vertebral artery and into the carotid artery. Thirty-nine of the 146 units were excited by the vertebral artery injections in the same time course as ventilatory augmentation. Eighteen of the 39 units did not react to peripheral chemoreceptor stimulation, i.e. they responded exclusively to central chemoreceptor stimulation. These 18 units were distributed in the caudal (‘C’) chemoceptive area of the ventral surface, in the vicinity of ventral respiratory group (VRG) neurons, and in the dorsal area ventral to the solitary tract. Twenty-one of the 39 units were excited by peripheral chemoreceptor stimulation, i.e. neurons with integrations of central and peripheral chemoreceptor inputs. They were densely packed in the nucleus paragigantocellularis lateralis, although also found in and around the rostral VRG neurons. Fifty-one of the 146 non-phasic units were excited (44) or inhibited (7) immediately after the vertebral artery injections, and their discharges returned to pre-injection levels prior to or during the early period of ventilatory augmentation. They also reacted to skin pinching, i.e. neurons with nociceptive action, and were found scattered variously in the ventral half of the medulla. Fourteen of the 146 units reacted to both chemoreceptor and nociceptor stimulations. The remaining 42 units were non-responsive. In summary, the distributions of the tonically active neurons excited by the central and/or peripheral chemoreceptors were not restricted to the ventral surface of the medulla.

Functional associations among simultaneously monitored lateral medullary respiratory neurons in the cat. I. Evidence for excitatory and inhibitory actions of inspiratory neurons.

Data were obtained from 45 anesthetized (Dial), paralyzed, artificially ventilated, bilaterally vagotomized cats. Arrays of extracellular electrodes were used to monitor simultaneously the activities of lateral medullary respiratory neurons located in the rostral and caudal regions of the ventral respiratory group. The average discharge rate as a function of time in the respiratory cycle was determined for each neuron and concurrent phrenic nerve activity. Most cells were tested for axonal projections to the spinal cord or the ipsilateral vagus nerve using antidromic stimulation techniques. Seven hundred and sixty-one pairs of ipsilateral respiratory neurons that contained at least one neuron whose maximum discharge rate occurred during the inspiratory phase were analyzed by cross-correlation of the simultaneously recorded spike trains. Twenty-three percent of the 410 pairs of inspiratory (I) neurons showed short time scale correlations indicative of functional association due to paucisynaptic connections or shared inputs. Eight per cent of the 351 pairs composed of an I cell and and expiratory (E) neuron were correlated. We found evidence for excitation of both bulbospinal I neurons and I cells that were not antidromically activated by stimulation of the spinal cord and vagus nerve (NAA neurons) by NAA I cells. We also obtained data suggesting inhibitory actions of cells whose maximum discharge rate occurred in the first half of the I phase (I-DEC neurons). These actions included inhibition of other I-DEC neurons, inhibition of cells whose greatest firing rate occurred in the last half of the I phase (I-AUG neurons), inhibition of E-DEC neurons, and inhibition of E-AUG cells. Sixty-two percent (31/50) of the correlations that could be interpreted as evidence for an excitatory or inhibitory paucisynaptic connection were detected in pairs composed of a caudal and a rostral ventral respiratory group neuron. Eighty-eight percent (14/16) of proposed intergroup excitatory connections involved a projection from the rostral neuron of the pair to the caudal cell, whereas 73% (11/15) of proposed inhibitory connections involved a caudal-to-rostral projection. These results support and suggest several hypotheses for mechanisms that may help to control the development of augmenting activity in and the timing of each phase of the respiratory cycle.