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Biomedical subjects

A L Bianchi

Publications and source records attributed to A L Bianchi.

At least 19 recordsLinked to original sources

Activity of respiratory laryngeal motoneurons during fictive coughing and swallowing.

Membrane potential changes and discharges from 28 laryngeal motoneurons were recorded intracellularly in the caudal nucleus ambiguus of decerebrate, paralyzed and ventilated cats. Electrical activities were recorded from 17 expiratory laryngeal motoneurons (ELMs) with maximal depolarizing membrane potential in early expiration, and from 11 inspiratory laryngeal motoneurons (ILMs) with maximal depolarizing membrane potential in inspiration. Activities during breathing were compared with those observed during fictive coughing and swallowing evoked by electrical stimulation of the superior laryngeal nerves. These non-respiratory behaviors were evidenced in paralyzed animals by characteristic discharge patterns of the phrenic, abdominal nerves and pharyngeal branch of the vagus nerve. We recorded the activity of 11 ELMs and 5 ILMs during coughing in which ELMs, but not ILMs, exhibited increased membrane depolarization and discharge frequencies. Membrane depolarization and discharge frequencies of all ELMs were also significantly increased during swallowing. In addition, membrane depolarization of most ELMs (15/17) was preceded by a short-lasting hyperpolarization due to chloride-dependent inhibitory mechanisms occurring at the onset of swallowing. Out of 10 ILMs tested during swallowing, 7 exhibited membrane depolarization, preceded in 5 cases by a short-lasting hyperpolarization. Discharge frequencies of ILMs were significantly reduced during swallowing. The same pattern of phasic activities of ILMs and ELMs was observed during coughing and breathing, suggesting the involvement of similar excitatory pathways in both behaviors. These results imply that the duration of activation and the discharge frequency of neurons of the central generator for breathing that drive laryngeal motoneurons are enhanced during coughing. During swallowing, in addition to central excitatory mechanisms, laryngeal motoneurons are subjected to an initial inhibition of unknown origin. This inhibition probably contributes to the temporal organization of the swallowing motor sequence.

Animals↗

Potent inhibition of both the acute and delayed emetic responses to cisplatin in piglets treated with GR205171, a novel highly selective tachykinin NK1 receptor antagonist.

The effects of GR205171, a selective tachykinin NK1 receptor antagonist, were investigated on both the acute and delayed phases of cisplatin-induced nausea-like behaviour and vomiting in the conscious piglet. Animals receiving cisplatin (5.5 mg kg(-1), i.v.) were observed for 60 h. Fifteen min prior to cisplatin infusion (T0(-15 min)), eight piglets acting as controls received an intravenous injection of saline solution (1 ml kg(-1)), whereas experimental animals received a single i.v. administration of GR205171 (1 ml kg(-1)) at a dose of 0.01 (n=8), 0.03 (n=8), 0.1 (n = 8), 0.3 (n = 16) or 1.0 (n = 13) mg kg(-1). In eight additional piglets, GR205171 (1 mg kg(-1)) was administered 15 min before the onset of the delayed phase (T16(-15 min)). A further five piglets received GR205171 (1 mg kg(-1)) every 6 h throughout the experiment. The latencies of the first emetic episode (EE) and nausea-like behavioural episode (NE) increased in all experimental groups treated at T0(-15 min), and the total number of both EE and NE during the 60 h was reduced in a dose-dependent manner. In piglets treated at T0(-15 min) with GR205171 1 mg kg(-1), eight out of 13 (62%) did not vomit throughout the experiment. Animals treated with GR205171 (1 mg kg(-1)) at T16(-15 min) exhibited an acute response to cisplatin but did not vomit during the delayed phase. The greatest inhibition of both nausea-like behaviour and vomiting was observed in piglets receiving multiple injections of GR205171. These results demonstrate the long-lasting anti-emetic effects of GR205171, and confirm the key role of substance P within the emetic reflex.

Animals↗

Differential brainstem Fos-like immunoreactivity after laryngeal-induced coughing and its reduction by codeine.

We used the expression of the immediate-early gene c-fos, a marker of neuronal activation, to localize brainstem neuronal populations functionally related to fictive cough (FC). In decerebrate, paralyzed, and ventilated cats, the level of Fos-like immunoreactivity (FLI) was examined in five groups of animals: (1) controls, sham-operated unstimulated animals; (2) coughing cats, including both animals in which FC was elicited by unilateral electrical stimulation of the superior laryngeal nerve (SLN) and (3) those in which FC was elicited by bilateral SLN stimulation; (4) stimulated-treated cats, in which bilateral SLN stimulation was applied after selective blockade of FC by codeine; and (5) codeine controls, sham-operated unstimulated cats subjected to administration of codeine. Fifteen brainstem structures were compared for numbers of labeled cells. Because codeine selectively blocks FC, brainstem nuclei activated specifically during FC were identified as regions showing increased FLI after FC and significant reductions in FLI after FC suppression by codeine in stimulated-treated cats. In coughing animals, we observed a selective immunoreactivity in the interstitial and ventrolateral subdivisions of the nucleus of the tractus solitarius, the medial part of the lateral tegmental field, the internal division of the lateral reticular nucleus, the nucleus retroambiguus, the para-ambigual region, the retrofacial nucleus, and the medial parabrachial nucleus. FLI in all these nuclei was significantly reduced in stimulated-treated cats. Our results are consistent with the involvement of neurons overlapping the main brainstem respiratory-related regions as well as the lateral tegmental field and the lateral reticular nucleus in the neural processing of laryngeal-induced FC.

Animals↗

Activity of dorsal respiratory group inspiratory neurons during laryngeal-induced fictive coughing and swallowing in decerebrate cats.

Membrane potential changes and/or discharges from 36 inspiratory neurons were recorded intracellularly in the dorsal respiratory group (DRG; i.e., the ventrolateral subdivision of the nucleus tractus solitarii) in decerebrate, paralyzed, and ventilated cats. Electrical activities were recorded from both somata (n = 10) and axons (n = 26). Activities during quiet breathing were compared with those observed during fictive coughing and swallowing evoked by repetitive electrical stimulation of afferent fibers of the superior laryngeal nerve (SLN). These nonrespiratory behaviors were evident in paralyzed animals as characteristic discharge patterns of the phrenic, abdominal, and hypoglossal nerves. Twenty-six neurons exhibiting antidromic action potentials in response to electrical stimuli applied to the cervical (C3-5) spinal cord were classified as inspiratory bulbospinal neurons (IBSNs). These neurons were considered as premotoneurons. The remaining 10 inspiratory neurons (I-NAA) were not antidromically activated by electrical stimuli applied to either cervical spinal cord or ipsilateral cervical vagus. These neurons are thought to be propriobulbar neurons. We recorded the activity of 31 DRG inspiratory neurons (24 IBSNs and 7 I-NAA) during coughing. All but one (a late-recruited IBSN) discharged a burst of action potentials during the coughing-related phrenic nerve activity. Typically, ramp-like membrane depolarization trajectories and discharge frequencies during coughing were similar to those observed during inspiration. We recorded the activity of 33 DRG inspiratory neurons (23 IBSNs and 10 I-NAA) during swallowing. Most (28/33) neurons were briefly activated, i.e., discharged a burst of action potentials during swallowing, but peak discharge frequency decreased compared with that measured during inspiration. The membrane potentials of nine somata exhibited a brief bell-shaped depolarization during swallowing, the amplitude of which was similar to that observed during inspiration. These results suggest that some inspiratory premotoneurons and propriobulbar neurons of the DRG might be involved in nonrespiratory motor activities, even if clearly antagonistic to breathing (e.g., swallowing). We postulate the existence in the medulla oblongata of adult mammals of neurons exhibiting a "functional flexibility".

Action Potentials↗

Brainstem neurons with projecting axons to both phrenic and abdominal motor nuclei: a double fluorescent labeling study in the cat.

The distribution of retrogradely doubly labeled brainstem neurons were analyzed in the cat after injection of two different fluorescent markers into the phrenic and abdominal motor nuclei. Diamidino Yellow (DY) was first injected either ipsilaterally or bilaterally into the ventral horn of lumbar spinal cord, and then Fast Blue (FB) into the right ventral horn of cervical spinal cord. Doubly labeled neurons were mainly found in the caudal ventrolateral medulla (retroambiguus region), in the dorsomedial and dorsolateral regions of the nucleus of the tractus solitarius (NTS) and in the raphe nuclei. In addition, doubly labeled neurons were found in the parabrachial and Kölliker-Fuse nuclei. Our results give anatomical evidence that pontine and medullary neurons are the source of a common pathway to both phrenic and abdominal motor nuclei. These neurons might be involved in strain efforts for expulsion such as vomiting or defecation.

Abdominal Muscles↗

Psychometric properties of the Brazilian Health Locus of Control Scale.

Based on social learning theory, the construct of health locus of control has proven valuable in predicting a wide variety of health-related behaviors. In studying this concept among Brazilians, the psychometric properties of the Multidimensional Health Locus of Control Scale translated into Brazilian Portuguese were investigated in a sample of 280 middle-class persons. Three types of health locus of control were verified, internal, powerful others, and chance. Further refinement of subscales is needed to improve internal consistency reliabilities.

Adult↗

[A simplified technique for decerebrating rats].

A simple and effective technique is described for decerebration of rat. The brain tissue at the precollicular level was gently and slowly aspirated. The two cut surfaces of the brainstem were covered with small pieces of gelfoam sponge or cotton balls soaked in thrombin solution. No ligation or clamp of the common carotid arteries of their branches was performed. After decerebration with this technique, the respiration rate, the heart rate and the mean femoral arterial pressure of the animals were stable for more than 10 hours. Successful intracellular recordings and labelings of medullary respiratory neurons were obtained from this type of preparation of rat. The technique for decerebrating rats described here may be used for acute experiments in which brainstem regulatory functions are investigated.

Animals↗

Intracellular electrophysiological and morphological study of the medullary inspiratory neurons of the decerebrate rat.

Intracellular recordings and labelings with horseradish peroxidase (HRP) of inspiratory neurons were performed in decerebrate, paralyzed and ventilated rats. A total of 58 neurons were located within the ventrolateral medulla. They were identified as bulbospinal neurons (n = 15), cranial motoneurons (n = 9) and not antidromically activated (NAA) neurons (n = 34) by antidromic stimulation or HRP labeling, or both. These inspiratory neurons had rhythmical changes in membrane potentials similar to those reported in cats, i.e. an abrupt depolarization at the onset of phrenic discharge followed by trajectories of depolarization which evolved into augmenting I, bell-shaped I or decrementing I patterns until a rapid repolarization at the start of expiration. All types were hyperpolarized during expiration by chloride-dependent inhibitory postsynaptic potentials (IPSPs) which were demonstrated in 13 neurons from which the reversal was obtained. Such IPSPs were apparent in two waves throughout expiration, an early one in post-inspiration (stage I of expiration) and a late one in late expiration (stage II of expiration). These properties are also similar to those of feline inspiratory medullary neurons. Four labeled bulbospinal neurons had axonal collaterals which were ipsi- and contralateral to the site of their somata. Two of 6 labeled NAA neurons exhibited profuse axonal arborizations within various medullary nuclei. No medullary axonal collateral was seen from 6 labeled motoneurons. These results indicate that even though in the rat a single concentration of inspiratory neurons within the ventrolateral medulla has been demonstrated, there is no fundamental difference in the organization of the inspiratory neuronal network compared to that of the cat.

Animals↗

Medullary expiratory neurons in the decerebrate rat: an intracellular study.

Intracellular recordings and labelings with horseradish peroxidase (HRP) of expiratory (E) neurons were performed in decerebrate, paralyzed, and ventilated rats. A total of 37 neurons were recorded, from which 4 cells and 1 axon were labeled. They were located in two regions of the ventrolateral medulla. One was in the rostral portion of the nucleus ambiguus just caudal to the facial nucleus, and the other in the nucleus retroambiguus at the level of the caudal medulla. These expiratory neurons had rhythmical changes in membrane potential similar to those reported in cat, i.e., a depolarization in the intervals between phrenic bursts which evolved in an augmenting (E-aug, n = 15), or bell-shaped or 'plateau' (E-all, n = 22) pattern until a rapid hyperpolarization at the start of inspiration. Both types were hyperpolarized during inspiration by chloride-dependent, inhibitory postsynaptic potentials (IPSPs) which were demonstrated in 17 neurons (10 E-aug and 7 E-all) from which reversal was obtained. Such IPSPs also existed during post-inspiration (stage I of expiration) in 4 of the 10 augmenting E neurons. They were identified by antidromic stimulation or HRP labeling, or both, as bulbospinal neurons (n = 2), cranial motoneurons (n = 4), or not antidromically activated (NAA) neurons (n = 31). All the identified bulbospinal neurons and the motoneurons exhibited an E-all pattern. The expiratory neurons of the caudal medulla had various projections as demonstrated with HRP labeling: one bulbospinal neuron with ipsilateral axon giving off intramedullary collaterals, and NAA neurons with rostral medullary projections or with axons crossing the midline.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Study of the topographical distribution of different populations of motoneurons within rat's nucleus ambiguus, by means of four different fluorochromes.

The topographical neuronal distribution within the rat nucleus ambiguus has been studied with the simultaneous retrograde labeling technique by means of four different fluorochromes injected within the various muscles and/or nerves of the oro-pharyngeal region. This technique has permitted the identification of several types of neurons along the same coronal plane. Most were motoneurons innervating the various muscles of the upper airway, including pharyngeal constrictor, stylopharyngeal, intrinsic laryngeal and the upper portion of the esophagus. Some neurons may have been preganglionic parasympathetic neurons. No evidence of axonal branching of any of the labeled motoneurons or parasympathetic neurons was found.

Animals↗

Discharge patterns of phrenic motoneurons during fictive coughing and vomiting in decerebrate cats.

In decerebrate, paralyzed, and ventilated cats, we recorded the activity of 100 spontaneously active phrenic motor axons during the increased phrenic discharges characteristic of fictive vomiting (FV) and coughing (FC). During control respiratory cycles, approximately one-half the neurons were recruited in the first decile of inspiration; recruitment continued throughout inspiration. During FV, the duration of phrenic discharge was halved; 20 of 26 motoneurons studied were recruited in the first decile of the burst. During FC, recruitment times did not change compared with control, although the duration of the phrenic burst doubled. Discharge frequencies increased and recruitment order of phrenic motoneurons was virtually unaffected during FC and FV. Limited recruitment of previously inactive neurons in the filaments from which we recorded was found during FV and FC. During FV, 1 previously inactive motoneuron was recruited in 16 filaments containing 25 spontaneously active motor axons. During FC, 3 new motoneurons were recruited in addition to the 64 already active in 35 filaments. Recruitment during FV and FC was absent even when recording from filaments known, on the basis of antidromic activation, to contain inactive motor axons. During FV, 10 of 26 motoneurons began their discharges with doublets (interspike interval < 10 ms); doublets occurred in only 4 of 67 motoneurons during FC. Already active phrenic motoneurons contributed to the intense phrenic activity associated with both respiratory (coughing) and nonrespiratory (vomiting) behavior by increases in discharge frequency, earlier recruitment, and doublets; the contribution of previously quiescent motoneurons remains uncertain.

Animals↗

Membrane potential changes of phrenic motoneurons during fictive vomiting, coughing, and swallowing in the decerebrate cat.

1. The patterns of membrane potential changes of phrenic motoneurons were compared during fictive vomiting, fictive coughing, and fictive swallowing in decerebrate, paralyzed cats. These fictive behaviors were identified by motor nerve discharge patterns similar to those recorded from the muscles of nonparalyzed animals. Phrenic motoneurons (n = 54) were identified by antidromic activation from the thoracic phrenic nerve. Intracellular recordings were obtained from 27 motoneurons during fictive vomiting, 40 during fictive coughing, and 27 during fictive swallowing. Sixteen motoneurons were recorded during both fictive coughing and fictive swallowing, eight during both fictive coughing and fictive vomiting, and two during both fictive vomiting and fictive swallowing. Seven motoneurons were studied during all three behaviors. 2. Fictive vomiting, typically evoked by electrical stimulation of abdominal vagal afferents, was characterized by a series of bursts of coactivation of phrenic and abdominal motor nerves, culminating in an expulsion phase in which abdominal discharge was prolonged both with respect to phrenic discharge and to abdominal discharge during the preceding retching phase. During fictive vomiting, phrenic motoneurons depolarized abruptly, and the amplitude of depolarization was significantly greater than during control inspirations. They then repolarized slowly throughout the phrenic burst, rapidly repolarizing at the end of each phrenic burst during retching and reaching a level similar to that observed during expiration. During the expulsion phase, the pattern was initially the same. However, after the cessation of phrenic discharge, the membrane potential repolarized slowly until the end of the abdominal burst, exhibiting greater synaptic noise than during expiration. One phrenic motoneuron, presumably innervating the periesophageal region of the diaphragm, received a strong hyperpolarization just before the onset of the emetic episode and fired for shorter periods during fictive vomiting than did other phrenic motoneurons.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Localization of respiratory bulbospinal and propriobulbar neurons in the region of the nucleus ambiguus of the rat.

The location of neurons within the ventral respiratory group (VRG) of rat was mapped following injections of 3 different fluorochrome tracers into different sites known to receive projections from VRG neurons. Injection sites included muscles innervated by the vagus (X) and glossopharyngeal (IX) nerves, and the sites of expiratory activity in the caudal medulla and of inspiratory activity in the spinal cord at the C4 level. Labeling of vagal motoneurons resulting from fluorochrome injections into muscles innervated by X and IX nerves was always ipsilateral to the site of injection. Both propriobulbar and bulbospinal neurons had primarily ipsilateral projections. No double-labeled cell bodies were observed. The cell bodies of the 3 types of neurons, propriobulbar, bulbospinal and vagal/glossopharyngeal, were unevenly distributed along the rostrocaudal axis of the VRG, suggesting a complex mosaic of neurons which regulate respiratory-related functions such as swallowing and vocalization.

Animals↗

Are the post-inspiratory neurons in the decerebrate rat cranial motoneurons or interneurons?

We examined the membrane potentials of 63 respiratory neurons in the ventrolateral medulla of decerebrate rats, whose trajectories had the characteristics of the post-inspiratory neurons, i.e. exhibiting hyperpolarization during inspiration, rapid depolarization at end-inspiration and progressive repolarization with a decrementing pattern during the intervals between phrenic bursts. Synaptic responses of 6 post-inspiratory neurons which were tested by stimulation of cervical vagus or superior laryngeal nerves were excitatory. Eleven of these 63 post-inspiratory neurons were labeled by intracellular injection of horseradish peroxidase (HRP). Ten of these 11 labeled neurons were motoneurons since their axons exited the medulla after joining the roots of cranial nerves. However, only one of these motoneurons was antidromically activated by stimulation of the ipsilateral cervical vagus nerve. We assumed that most of the post-inspiratory medullary neurons of the present study were motoneurons, but not interneurons, although antidromic invasion was not possible after stimulation of the cervical vagus and superior laryngeal nerves. Two post-inspiratory neurons of this sample had bulbospinal axons, which were revealed by antidromical activation of spinal cord and HRP labeling, respectively. The axon of the labeled bulbospinal neuron had axonal collaterals which were distributed within the region of the nucleus ambiguous of the ipsilateral medulla. The functional significance of this type of post-inspiratory neuron is discussed.

Animals↗

Patterns of membrane potentials and distributions of the medullary respiratory neurons in the decerebrate rat.

We analyzed the membrane potential of 161 respiratory neurons in the medulla of decerebrate rats which were paralyzed and ventilated. Three types of inspiratory (I) neurons were observed: those displaying progressive depolarization in inspiration (augmenting I neurons), those which gradually repolarized after maximal depolarization at the onset of inspiration (decrementing I neurons) and those exhibiting a plateau or bell-shaped membrane potential trajectory throughout inspiration (I-all neurons). Three types of expiratory (E) neurons were also encountered: those in which the membrane potential progressively depolarized (augmenting E neurons), those in which the membrane potential repolarized during the interval between phrenic bursts (decrementing E or post-I neurons) and those exhibiting a plateau or bell-shaped membrane potential trajectory throughout expiration (E-all neurons). Axonal projections of these medullary neurons were identified in the cranial nerves (n = 34), or in the spinal cord (n = 19) as revealed by antidromic stimulation and/or by reconstruction following horseradish peroxidase (HRP) labeling. The other 108 neurons were not antidromically activated (NAA) by the stimulations tested, or had their axons terminating inside the medulla as revealed by HRP labeling. All these respiratory neurons, except for 3 which were hypoglossal motoneurons, had their somata within the ventrolateral medulla, in the region of the nucleus ambiguus, homologous to the ventral respiratory group (VRG) of the cat. No dorsal respiratory group (DRG) was detected within the medulla of the rats. Due to this absence of a DRG, it is concluded that the neural organization of respiratory centers is quite different in cats and rats.

Animals↗

Activity of respiratory-related oropharyngeal and laryngeal motoneurones during fictive vomiting in the decerebrate cat.

Activities of respiratory laryngeal and oropharyngeal respiratory nerves were studied during fictive vomiting elicited by supradiaphragmatic vagus nerve stimulation in the decerebrate cat. Inspiratory laryngeal nerves were strongly inhibited throughout the retching and expulsion phase. Glossopharyngeal, hypoglossal and expiratory laryngeal nerves were coactivated with the phrenic and abdominal nerve bursts. The pharyngeal branch of the vagus nerve discharged during the phrenic and abdominal inter-burst of the retching phase, and was silent during the abdominal expulsion. These activities permit speculation about the role of upper airway muscles during vomiting.

Action Potentials↗

Discharge patterns of laryngeal motoneurones in the cat: an intracellular study.

In decerebrate cats, stable intracellular recordings were made from 37 laryngeal motoneurones, the membrane potentials of which varied in relation to respiration. These motoneurones were identified as laryngeal since all were antidromically activated by stimulation of the recurrent laryngeal nerve, but in two, the antidromic activity could only be elicited by vagal stimulation (vagotomized cats). The cell bodies were all located within the nucleus ambiguous. Sixteen cells were depolarized during the phrenic burst and were classified as inspiratory laryngeal motoneurones (ILM). They repolarized at end-inspiration and received two successive waves of postsynaptic inhibition during expiration: an early, strong one and a late (end-expiratory), weaker one. The decay of the first wave was related to the duration of postinspiratory phrenic activity. Twenty-one cells depolarized abruptly in early expiration followed by a more-or-less gradual repolarization. They were classified as expiratory laryngeal motoneurones (ELM). All ELM were strongly inhibited during inspiration. Some of them received weak inhibition during end expiratory phase. The rapid and large depolarization observed during early expiration (and consequent maximal discharge frequency) can be explained by two summating mechanisms: a postinhibitory rebound resulting from the removal of inhibition during inspiration, and an excitatory phenomenon of unknown origin. The amplitude of this excitatory phenomenon was largest in cats with the most residual (early expiratory) phrenic activity. To explain the hyperpolarizations occurring in ELM during late expiratory and inspiratory phases and those occurring in ILM during early expiration, we hypothesize that reciprocal inhibition exists between networks controlling ILM and ELM activities or between these motoneurones themselves.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗