Axonal projections of medullary swallowing neurons in guinea pigs.

A central pattern generator (CPG) for swallowing in the medulla oblongata generates spatially and temporally coordinated movements of the upper airway and alimentary tract. To reveal the medullary neuronal network of the swallowing CPG, we examined the cytoarchitecture of the swallowing CPG and axonal projections of its individual neurons by extracellular recording and juxtacellular labeling of swallowing-related neurons (SRNs) in the medulla in urethane-anesthetized and paralyzed guinea pigs. Three major types of neuronal discharge patterns were identified during fictive swallowing induced by stimulation of the superior laryngeal nerve: early (burst-like activation during the pharyngeal stage), late (activation after the pharyngeal stage), and inhibited (inhibition during the pharyngeal stage) types.

Respiration-related afferents to parabrachial pontine regions.

The dorsolateral pons around the parabrachial nucleus is an important participant in respiratory control. This area involves various respiration-related neurons, and their respiratory modulation is thought to arise from afferents from medullary respiratory neurons. Today, however, only a limited number of afferent sources have been identified.

Overall distribution of GLYT2 mRNA-containing versus GAD67 mRNA-containing neurons and colocalization of both mRNAs in midbrain, pons, and cerebellum in rats.

We aimed to clarify the overall distribution of glycinergic neurons in the midbrain, pons, and cerebellum in rats, using in situ hybridization for mRNA encoding glycine transporter 2 (GLYT2), which reliably detects glycinergic cell bodies. We combined this method with in situ hybridization for mRNA encoding glutamic acid decarboxylase isoform 67 (GAD67), and have presented for the first time global and detailed views of the distribution of glycinergic neurons in relation to GABAergic neurons. In addition to this single-detection study, we performed double-detection of GLYT2 mRNA and GAD67 mRNA to determine the distribution of neurons co-expressing these mRNAs.

Reflections on respiratory rhythm generation.

Knowledge about neuronal mechanisms that control respiration is being advanced rapidly by studies that make use of both mature in vivo animals and in vitro neonates. The available data suggest that particular types of neurons within selected networks of the ventrolateral medulla are essential for respiratory rhythm generation. There are many uncertainties, however, about the correspondence between neurons identified by the above two approaches, because there are virtually no studies that have combined them.

A review of functional and structural components of the respiratory center involved in the arousal response.

State-dependent changes influencing both the central chemoreceptor and vagal inputs to respiratory neurons may provide useful markers to assess some intrinsic factors of the respiratory center. In this paper we discuss the following topics from our recent experiences, and their relevance to the assessment of sleep-related phenomena. (1) 'Post-sigh' apnea appears predominantly during non-REM sleep in control subjects, and is a potential marker of respiratory dysfunction during this stage of sleep.

Glycine is used as a transmitter by decrementing expiratory neurons of the ventrolateral medulla in the rat.

The medullary respiratory network involves various types of respiratory neurons. The present study focused on possible inhibitory neurons called decrementing expiratory (E-DEC) neurons and aimed to determine whether their transmitter is glycine or GABA. In Nembutal-anesthetized, neuromuscularly blocked, and artificially ventilated rats we labeled E-DEC neurons with Neurobiotin and processed the tissues for detection of mRNA encoding either glycine transporter 2 (GLYT2) as a marker for glycinergic neurons or glutamic acid decarboxylase isoform 67 (GAD67) as a marker for GABAergic neurons, using in situ hybridization.

Distribution of glycine transporter 2 mRNA-containing neurons in relation to glutamic acid decarboxylase mRNA-containing neurons in rat medulla.

We studied the distribution of medullary glycinergic neurons in relation to GABAergic neurons, by using in situ hybridization method for mRNA encoding either glycine transporter 2 (GLYT2) or glutamic acid decarboxylase isoform 67 (GAD67). GLYT2 mRNA-positive (GLYT2+) neurons were distributed widely and clustered in (1). the respiration-related area of the ventrolateral medulla called the Bötzinger complex, (2).

Activity of neurons in ventrolateral respiratory groups during swallowing in decerebrate rats.

To elucidate the neuronal basis of the coordination between swallowing and respiration, we examined the swallowing-related activity of respiratory neurons in the ventrolateral respiratory groups of the medulla oblongata of decerebrate, paralyzed and artificially ventilated rats (n = 14). Extracellular recording was made during fictive swallowing evoked by the electrical stimulation of the superior laryngeal nerve from a total of 141 neurons with respiratory rhythm (99 expiratory and 42 inspiratory neurons). The burst of discharge by the hypoglossal nerve was used to monitor the pharyngeal phase of swallowing.

Activity of brainstem respiratory neurones just before the expiration-inspiration transition in the rat.

Inspiratory activity of the hypoglossal nerve (XIIn) often precedes that of the phrenic nerve (PHRn). By manipulating artificial respiration, this preceding activity (pre-I XIIn activity) can be lengthened or isolated prematurely (decoupled XIIn activity) without developing into overt PHRn-associated inspiratory bursts. We hypothesized that these pre-I and decoupled XIIn activities, collectively termed 'XIIn-w/o-PHRn activity', reflect certain internal states of the respiratory centre at the period just prior to the transition from the expiratory phase to the inspiratory phase.

Brainstem and spinal projections of augmenting expiratory neurons in the rat.

There are two types of expiratory neurons with augmenting firing patterns (E-AUG neurons), those in the Bötzinger complex (BOT) and those in the caudal ventral respiratory group (cVRG). We studied their axonal projections morphologically using intracellular labeling of single E-AUG neurons with Neurobiotin, in anesthetized, paralyzed and artificially-ventilated rats. BOT E-AUG neurons (n = 11) had extensive axonal projections to the brainstem, but E-AUG neurons (n = 5) of the cVRG sent axons that descended the contralateral spinal cord without medullary collaterals.

Intracellular activity of superior laryngeal nerve motoneurons during fictive swallowing in decerebrate rats.

We examined the swallowing-related intracellular activity of motoneurons of the superior laryngeal nerve (SLN) in decerebrate, paralyzed and artificially-ventilated rats, to elucidate the neuronal mechanism of the pharyngo-esophageal and laryngo-esophageal coordination during swallowing. The majority of the SLN motoneurons exhibited respiratory rhythm (n=16; 13 inspiratory, one expiratory and two non-respiratory neurons). During fictive swallowing evoked by electrical stimulation of the SLN, all these motoneurons showed a hyperpolarization-depolarization sequence in their membrane potentials.

Difference between hypoglossal and phrenic activities during lung inflation and swallowing in the rat.

We aimed in this study to elucidate the discharge properties and neuronal mechanisms of the dissociation between hypoglossal and phrenic inspiratory activities in decerebrate rats, which had been subjected to neuromuscular blockade and artificially ventilated. The discharge of the hypoglossal nerve and the intracellular activity of hypoglossal motoneurones were monitored during respiration and fictive-swallowing evoked by electrical stimulation of the superior laryngeal nerve, and were compared with the activity of the phrenic nerve. The hypoglossal nerve activity was characterized by its onset preceding the phrenic nerve activity ('pre-I' activity).

Morphology of the decrementing expiratory neurons in the brainstem of the rat.

In anesthetized and artificially-ventilated rats, the morphological properties of decrementing expiratory (E-DEC) neurons were studied using intracellular recording and labeling with Neurobiotin. Sixteen E-DEC neurons were successfully labeled; ten of which were cranial motoneurons located in the facial (FN) and ambiguus (NA) nuclei. Two interneurons were labeled in the Bötzinger complex (BOT) and the ventral respiratory group (VRG) rostral to the obex, and the remaining four in the VRG caudal to the obex.

Swallowing-related activities of respiratory and non-respiratory neurons in the nucleus of solitary tract in the rat.

Swallowing-related activity was examined in respiratory (n = 60) and non-respiratory (n = 82) neurons that were located in and around the nucleus of the solitary tract (NTS) in decerebrated, neuromuscularly blocked and artificially ventilated rats. Neurons that were orthodromically activated by electrical stimulation of the superior laryngeal nerve (SLN) were identified, and fictive swallowing was evoked by SLN stimulation. The pharyngeal phase of swallowing was monitored by hypoglossal nerve activity.

Axonal projections of pulmonary slowly adapting receptor relay neurons in the rat.

We elucidated efferent projections of second-order relay neurons (P-cells) activated by afferents originating from slowly adapting pulmonary receptors (SARs) to determine the central pathway of the SAR-evoked reflexes. Special attention was paid to visualizing the P-cell projections within the nucleus tractus solitarii (NTS), which may correspond to the inhibitory pathway from P-cells to second-order relay neurons (RAR-cells) of rapidly adapting pulmonary receptors. P-cells were recorded from the NTS in Nembutal-anesthetized, paralyzed, and artificially ventilated rats.