Primary graft dysfunction after liver transplantation: from pathogenesis to prevention

Spontaneous bladder contractions (SBCs) in decerebrate, vagotomized, paralyzed, ventilated cats have been shown to decrease phrenic and hypoglossal inspiratory nerve activities, as well as the activities of other respiratory motor nerves. To determine whether vagal afferents from the lung influence the respiratory inhibition associated with SBCs, we recorded phrenic and hypoglossal nerve activities in decerebrate, paralyzed, vagally intact cats. The animals were ventilated by a servo-respirator, which inflated the lungs in accordance with integrated phrenic nerve activity. Maintained increases in end-expiratory lung volume were produced by the application of 2-10 cm H2O positive end-expiratory pressure (PEEP). SBCs were accompanied by decreases in both phrenic and hypoglossal peak integrated nerve activities, as well as by marked decreases in respiratory frequency. The reduction of respiratory frequency was greater with higher levels of PEEP, a few animals becoming apneic during SBCs. After bilateral vagotomy, SBCs continued to decrease phrenic and hypoglossal peak integrated nerve activities as previously reported, but the reduction of respiratory frequency was much less striking than when the vagi were intact. These results indicate that activity of vagal afferents from the lung augments the respiratory influence of SBCs. Furthermore, SBCs in vagally intact animals can induce periodic breathing.     

Changes in serum pepsinogen, gastrin, and immunoglobulin G antibody titers in helicobacter pylori-positive gastric ulcer after eradication of infection

The feasibility of using the spiral nerve cuff electrode design for recordings of respiratory output from the hypoglossal (HG) and phrenic nerves is demonstrated in anesthetized, paralyzed, and artificially ventilated cats. Raw neural discharges of the HG nerve were analyzed in terms of signal-to-noise ratios and frequency spectra. The rectified and integrated moving average activity of the HG nerve had a peak value of 1.74 +/- 0.21 microV and a baseline value of 0.72 +/- 0.11 microV at elevated respiratory drive induced by increases in CO2 or oxygen deprivation when recorded with 10-mm-long cuffs. The frequency content of the HG electroneurogram extended from several hundred hertz to 6 kHz. Spiral nerve cuff recordings without desheathing of the nerve provided large enough signal-to-noise ratios that allowed them to be used as a measure of respiratory output and had much wider frequency bandwidths than the hook electrode preparations. A major advantage of the cuff electrode over the hook electrode was its mechanical stability, which significantly improved the reproducibility of the recordings both in terms of signal amplitudes and frequency contents.     

The effects of caffeine on the respiratory depression by morphine

The effects of intravenous administration of caffeine on the discharge of the phrenic nerve were studied following vagotomy in 7 pentobarbital anesthetized mechanically ventilated rats. Morphine (0.4 mg.kg-1.min-1) was administered until the respiratory rate decreased to about half of the baseline respiratory rate. In those state, we first administered caffeine (20 mg.kg-1), intravenously and then administered naloxone (0.02 mg) intravenously. The increase of inspiratory time from 0.49 +/- 0.16 to 2.01 +/- 0.47 s by morphine recovered to 0.86 +/- 0.38 s by caffeine and 0.50 +/- 0.22 s by naloxone. Expiratory time did not change during each drug administration. The decrease of respiratory rate from 46.6 +/- 5.9 to 20.6 +/- 4.1 breaths.min-1 by morphine recovered to 39.6 +/- 6.1 breaths.min-1 by caffeine and 47.6 +/- 4.6 breaths.min-1 by naloxone. Amplitude of integrated phrenic nerve discharge increased to 117 +/- 32% by caffeine and 156 +/- 39% by naloxone compared to the baseline. These results suggest that caffeine acts as a respiratory stimulant on the respiratory depression by morphine.     

Progeroid syndrome with characteristic facial appearance and hand anomalies in father and son

The effects of microinjection of sodium glutamate and glycine into ventrolateral nucleus of tractus solitarius (VLNTS) on the discharges of phrenic nerve were observed on 40 anaesthetized, vagotomized, paralyzed and artificially ventilated rabbits. The results were as follows: Phrenic nerve discharges were increased, inspiratory duration prolonged, expiratory duration shortened and respiratory frequency not changed by microinjection of sodium glutamate into VLNTS. While by microinjection of glycine, phrenic nerve discharges were decreased, even abolished, inspiratory duration shortened, expiratory duration irregularly prolonged and respiratory frequency decreased. The above results show that VLNTS exert important effects on the genesis of respiratory rhythm.     

Respiratory activity of the rat posterior cricoarytenoid muscle

An anatomic and electrophysiological study of the rat posterior cricoarytenoid (PCA) muscle is described. The intramuscular nerve distribution of the PCA branch of the recurrent laryngeal nerve was demonstrated by a modified Sihler's stain. The nerve to the PCA was found to terminate in superior and inferior branches with a distribution that appeared to be confined to the PCA muscle. Electromyography (EMG) recordings of PCA muscle activity in anesthetized rats were obtained under stereotaxic control together with measurement of phrenic nerve discharge. A total of 151 recordings were made in 7 PCA muscles from 4 rats. Phasic inspiratory activity with a waveform similar to that of phrenic nerve discharge was found in 134 recordings, while a biphasic pattern with both inspiratory and post-inspiratory peaks was recorded from random sites within the PCA muscle on 17 occasions. The PCA EMG activity commenced 24.6 +/- 2.2 milliseconds (p < .0001) before phrenic nerve discharge. The results are in accord with findings of earlier studies that show that PCA muscle activity commences prior to inspiratory airflow and diaphragmatic muscle activity. The data suggest that PCA and diaphragm motoneurons share common or similar medullary pre-motoneurons. The earlier onset of PCA muscle activity may indicate a role for medullary pre-inspiratory neurons in initiating PCA activity.     

Effects of halothane on the phrenic nerve responses to carbon dioxide mediated by carotid body chemoreceptors in vagotomized dogs

BACKGROUND: Previous studies in dogs showed that the phrenic nerve response to an acute hypoxic stimulus was dose dependently depressed by 0.5-2.0 minimum alveolar concentration (MAC) of halothane but not abolished. Because a carbon dioxide stimulus is transduced by a different mechanism in the carotid body chemoreceptors (CBCRs) than is a hypoxic stimulus, inhalational anesthetics may preferentially depress one of these transduction processes, the central neuronal processing, or both, of the integrated responses to these two types of inputs. METHODS: Carotid body chemoreceptor stimulation was produced by short (1-1.5 s), bilateral, 100% carbon dioxide in saline infusions into the carotid arteries during neural inspiration in unpremedicated, halothane-anesthetized, paralyzed, vagotomized dogs during constant mechanical ventilation. The phrenic neurogram quantified the neural inspiratory response. Four protocols were performed in the study: (1) the dose-dependent effects of halothane anesthesia (0.5-2.0 MAC) during hyperoxic hypercapnia on phrenic nerve activity, (2) the effects of three background levels of the partial pressure of carbon dioxide (PaCO2) on the magnitude of the carbon dioxide infusion responses at 1 MAC halothane, (3) the effects of anesthetic type on the magnitude of the carbon dioxide infusion response, and (4) the effects of CBCR denervation. RESULTS: Peak phrenic nerve activity (PPA) increased significantly during the carbon dioxide-stimulated phrenic burst in protocols 1-3; after denervation there was no response (protocol 4). Halothane produced a dose-dependent reduction in the PPA of control and carbon dioxide infusion-stimulated phrenic bursts and in the net carbon dioxide response. The net PPA responses for the different PaCO2 background levels were not different but were somewhat larger for sodium thiopental anesthesia than for 1.0 MAC halothane. CONCLUSIONS: The phrenic nerve response to an acute, severe carbon dioxide stimulus was dose dependently depressed by surgical doses of halothane. The observed responses to carbon dioxide infusion were mediated by the CBCRs because they were eliminated by CBCR denervation. These results suggest that the CBCR transduction and central transmission of the carbon dioxide signal in terms of inspiratory excitatory drive are not abolished at surgical levels of halothane anesthesia.     

Ultrastructural changes in the rat phrenic nucleus developing within 2 h after cervical spinal cord hemisection

The present study was conducted to describe the ultrastructural changes which occur in the young adult rat phrenic nucleus within 2 h after an ipsilateral C2 spinal cord hemisection. The main objective was to determine if there is a temporal relationship between specific ultrastructural changes in the phrenic nucleus and a significant augmentation of crossed phrenic nerve activity which occurs as early as 2 h after hemisection. Phrenic motoneurons were identified at electron microscopic levels by retrograde HRP labeling. Ultrastructural features in the phrenic nucleus of control and experimental rats were qualitatively analyzed and then quantitated. At 2 h posthemisection, there was a significant increase in the mean percentage of phrenic dendrodendritic appositions. In the control rats, 4.73 +/- 0.18% of phrenic dendrites were in apposition, and this percentage increased significantly to 8.58 +/- 0.54% at 2 h after injury. Furthermore, the mean lengths of asymmetrical and symmetrical synaptic active zones increased significantly at 2 h posthemisection from control lengths of 0.372 +/- 0.009 microns and 0.404 +/- 0.007 microns to 0.410 +/- 0.011 microns and 0.513 +/- 0.032 microns, respectively, in experimental rats. The phrenic nucleus is therefore capable of morphological plasticity as early as 2 h after spinal cord hemisection and this plasticity coincides temporally with the physiological augmentation of crossed phrenic nerve activity at 2 h. The data further suggest that these morphological changes may be part of the substrate for the unmasking of ineffective synapses during the crossed phrenic phenomenon.     

Effects of carotid Body chemoreceptor stimulation by 5-HT on phrenic nerve activity and ventilation in the rabbit

Effects of carotid body chemoreceptor stimulation by 5-hydroxytryptamine (5-HT) on the phrenic nerve activity and ventilation were studied by injecting it into the external carotid artery of the rabbits. 5-HT induced an immediate and transient increase of ventilatory rate which became more pronounced with increasing the dose, but it was accompanied by decreases in integrated phrenic nerve activity and tidal volume. The excitatory response was followed by an inhibition in 35% of the experiments. The 5-HT-induced excitation was unaffected by pretreatment with atropine and mecamylamine, but it was completely blocked by tetrodotoxin (TTX) which failed to inhibit the excitation induced by NaCN. Prior administration of atropine abolished the inhibitory effect of 5-HT and the ventilatory excitation induced by baroreceptor stimulation. It may be suggested that the excitation induced by 5-HT results from stimulation of chemoreceptor nerve endings, whereas the inhibition is probably due to its action on baroreceptor nerve endings.     

Effects of intraphrenic injection of potassium on diaphragm activation

The purpose of the present study was to determine whether potassium, injected into the arterial supply of the diaphragm, would reflexly alter efferent diaphragmatic motor outflow and systemic arterial pressure. Studies were performed using in situ canine diaphragm muscle strips in which the inferior phrenic artery and vein were cannulated and all other sources of strip blood flow were ligated. Injection of potassium (0.1 meq) into the inferior phrenic artery elicited a small transient (1-2 breaths) decrease in the peak strip tension developed during spontaneous muscle contractions, in peak integrated strip electromyographic (EMG) activity, and in the peak integrated EMG activity of the contralateral hemidiaphragm. This was followed by a more pronounced and more sustained increase in each of these parameters as well as an increase in systemic arterial pressure. This latter excitatory response was qualitatively similar to that induced by the injection of capsaicin (5 and 25 micrograms) into the phrenic artery. Section of the left phrenic nerve abolished the effects of intra-arterial potassium and capsaicin on systemic arterial pressure and right hemidiaphragm EMG activity. These data support the existence of a potent excitatory phrenic-to-phrenic reflex that can be activated by potassium injection into the diaphragm. Activation of this pathway increases diaphragm motor activation and augments systemic arterial pressure.     

Transient respiratory augmentation elicited by acute head-down tilt in the anesthetized cat

Acute head-down tilt (AHDT, -30 degrees) in humans induces a transient ventilatory augmentation for 1-2 min accompanied by a high venous return. However, the mechanisms underlying this respiratory response remain obscure because of limitations of experiments carried out in human subjects. The present study was undertaken to determine whether AHDT-induced respiratory augmentation exists in the anesthetized, paralyzed, and ventilated cat and, if so, whether this response depends on 1) the cerebellum, 2) the carotid sinus (CS) and/or vagal afferents, and 3) elevation of central venous return. The integrated phrenic neurogram, arterial blood pressure, central venous pressure (CVP), and end-tidal PCO2 were recorded before, during, and after AHDT. The results showed that AHDT produced a transient ( approximately 2 min) enhancement of minute phrenic activity (approximately 30%) primarily via an increase in peak integrated phrenic neurogram amplitude associated with a remarkable elevation of CVP (approximately 3 min). Cerebellectomy, CS denervation, bilateral vagotomy, or clamping CVP did not affect the presence of the AHDT-induced minute phrenic activity response. These findings demonstrate that the anesthetized cat is a suitable model for investigating the mechanisms involved in AHDT-induced respiratory augmentation. Preliminary studies suggest that this response does not require the cerebellum, CS/vagal afferents, or an associated rise in central venous return.     

Medullary respiratory neuronal activity modulated by stimulation of the fastigial nucleus of the cerebellum

The ability of the rostral fastigial nucleus (FNr) of the cerebellum to modulate medullary respiratory neuronal activity was examined in 17 anesthetized, paralyzed and ventilated cats. A bipolar stimulating electrode was positioned into the FNr and tungsten microelectrodes used to record units within the nucleus tractus solitarius (NTS), nucleus ambiguus (NA) and nucleus retroambigualis (NRA). Transient stimuli (< 150 microA, 5-200 Hz) were delivered during inspiration or expiration, and the effects noted on medullary neuronal activity and the phrenic neurogram. The results showed that FNr stimulation: (1) modulated inspiratory and expiratory neuronal (ramp-, early- and late-inspiratory and stage I and II expiratory) discharges recorded from the NTS, NA and NRA (n = 67, 14 and 28) when stimuli (> or = 20-50 Hz) were delivered during either the inspiratory or expiratory phases; (2) terminated the burst durations of inspiratory (77%) and expiratory (94%) neurons with stimulus-response latencies of 28.2 +/- 3.1 ms (inspiratory) and 29.4 +/- 3.6 ms (expiratory); (3) elicited changes in phrenic neurogram concomitant with the effects noted on medullary neuronal activities; (4) failed to change heart rate and arterial blood pressure; and (5) did not affect medullary neuronal and phrenic nerve activity following kainic acid injection into the FNr. We conclude that activation of the FNr (likely its cell bodies) can modulate the respiratory output via influences on medullary respiratory-related neurons. The primary cerebellar effect across all sub-types of respiratory neurons was early termination.     

Diaphragm activation with intramuscular stimulation in dogs

We studied in 10 supine anesthetized dogs diaphragm contraction produced by electrical activation with intramuscular electrodes surgically implanted in the ventral surface of the diaphragm and compared this with activation of the ipsilateral phrenic nerve (C5, 6, and 7) before it entered the thorax. Repetitive 40-Hz pulse trains with supramaximal current stimulus were used after hyperventilation of the animals to apnea. A single intramuscular electrode within 1 to 2 cm of the site of phrenic nerve entry into the diaphragm produced a mean transdiaphragmatic pressure of 12.0 cm H2O +/- 0.97 SE and mean tidal volume of 0.27 L +/- 0.04 SE. Mean values observed with phrenic nerve stimulation were not statistically different, and both electrode systems produced equivalent outward abdominal motion and upper rib cage paradox, as monitored by inductive plethysmography. There was no difference in gas exchange during stimulation with a single hemidiaphragm electrode and mechanical ventilation compared at the same tidal volume and respiratory rate. Blockade of neuromuscular transmission with curare eliminated intramuscular and phrenic nerve stimulation proportionately, suggesting that activation of the diaphragm is dependent in both cases on the phrenic nerve. This technique does not entail manipulation of the phrenic nerve and may have clinical application as an alternative technique for diaphragm pacing.     

5-HT-1A receptor-mediated modulation of medullary expiratory neurones in the cat

The involvement of the 5-HT-1A receptor in serotoninergic responses of stage 2 expiratory (E-2) neurones was investigated in pentobarbitone-anaesthetized, mechanically ventilated cats. The specific agonist of the 5-HT-1A receptor, 8-hydroxy-diproplaminotetralin (8-OH-DPAT), administered systemically or by ionophoresis directly on to the neurones, had a clear depressant effect. Administration of 8-OH-DPAT at doses of 10-50 micrograms kg-1 (I.V.) increased the membrane hyperpolarizations of E-2 neurones during the inspiratory and postinspiratory phases, and shortened their duration of activity in association with shortening of phrenic nerve activity. Discharges of E-2 neurones were also less intense. At doses of 50-90 micrograms kg-1, 8-OH-DPAT reduced or abolished inspiratory hyperpolarizations, and reduced expiratory depolarizations of membrane potential and discharge in parallel with inhibition of phrenic nerve discharges. The effects of the larger doses were reversed by I.V. injection of NAN-190, an antagonist at the 5-HT-1A receptor. Dose-dependent effects on the membrane potential and discharge of E-2 neurones, but not on phrenic nerve activity, were also seen by ionophoretic administration of 8-OH-DPAT on to E-2 neurones. At low currents, ejection of 8-OH-DPAT hyperpolarized the neurones without affecting the duration of inspiratory hyperpolarization and expiratory depolarization. This hyperpolarization depressed the intensity and the duration of expiratory discharges. Ejection with larger currents hyperpolarized the E-2 neurones further, and depressed expiratory depolarization leading to blockade of expiratory discharges. The effects on membrane potential were accompanied by decreased neuronal input resistance. This depressed the excitability of E-2 neurones as tested by discharge evoked by intracellular current injection. The amplitudes of action potentials decreased in parallel with the changes in input resistance. The effects were attributed to a postsynaptic effect of 8-OH-DPAT leading to a gradually developing inhibition by activation of 5-HT-1A receptors. Hyperventilatory apnoea depressed on-going synaptic activity and unmasked the effect of ionophoretically applied 8-OH-DPAT. The responses of the E-2 neurone were enhanced, as evidenced by increased membrane hyperpolarization and greater reduction of input resistance. Both responses faded appreciably, indicating receptor desensitization. The degree and rate of apparent desensitization depended on the dose/ejecting current. The greater sensitivity and faster desensitization to 8-OH-DPAT were attributed to the hyperventilatory alkalinization of the extracellular fluid, which might influence agonist binding to 5HT-1A receptors and/or receptor properties.     

The pathophysiology of REM-sleep behavior disorder

Congenital partial pericardial defect is a rare anomaly that causes no symptoms and is often noticed by chance at autopsy or thoracotomy. During an operation on a patient with bronchiectasis, a partial pericardial defect and anomaly of left phrenic nerve were found incidently. A 58-year-old man complaining hemoptysis was referred to our hospital for surgical treatment of the left cystic bronchiectasis. During a thoracotomy, a partial pericardial defect was noticed. Moreover the left phrenic nerve could not be found within the operative field. We performed left pneumonectomy without repair of pericardial defect, and the patient had a satisfactory postoperative course. A relationship was suggested between congenital pericardial defect and the anomaly of the phrenic nerve.     

Pressure-independent inhibition of sympathetic activity by noradrenaline: role of baroreceptor C fibres

The effect of i.v. infusion of noradrenaline on activity in the renal sympathetic nerve was studied in rabbits anesthetized with chloralose and urethane. Noradrenaline (3--8 microgram/kg-min) initially increased mean arterial pressure 20--40 mmHg and consequently reduced renal nerve activity. However, studies over a wide range of pressures--obtained by changing the blood volume, revealed that noradrenaline after a few minutes had induced a pressure-independent reduction of sympathetic discharge. The effect disappeared with baroreceptor denervation. An unchanged relationship between arterial pressure and integrated activity in the whole left aortic nerve (which is largely a measure of activity in A fibres) suggested that the sympathetic depression was due to excitation of aortic nerve C fibres. This conclusion was supported by studies of sympathetic responses to selective stimulation of aortic nerve A and C fibres at equal pressures before and during infusion of noradrenaline. Compared to the reflex activity from A fibres, C fibre stimulation was invariably less effective in suppressing renal nerve activity during the infusion. Our studies indicate that noradrenaline may effect a negative feedback control of sympathetic discharge through activation of baroreceptor C fibres.     

Neural respiratory responses to cortically induced seizures in cats

Seizure activity can lead to profound respiratory stimulation in spontaneously breathing animals with intact respiratory feedback mechanisms (Paydarfar et al., Am. J. Physiol. 260, R934, 1991). The present study was designed to test the hypothesis that peripheral respiratory feedback mechanisms are not important for the genesis of seizure-induced hyperpnea. Studies were performed in 16 anesthetized, vagotomized, glomectomized cats whose end-tidal PCO2 (PETCO2) was kept constant. Integrated phrenic nerve activity was used to represent respiration. Seizures were induced by injection of penicillin into the parietal cortex and electrocorticographic (ECoG) and biceps femoris nerve activities, arterial pressure, airway PCO2 and brain temperature were recorded continuously. Progressive seizure activity was associated with progressive increases of respiratory frequency and peak phrenic activity, despite constancy of PETCO2 and brain temperature. Patterns of entrainment were identified among ECoG spikes, biceps femoris nerve and phrenic nerve activities. Phrenic nerve activity became highly irregular during generalized ictal seizures and ceased to respond to changes of PETCO2. Acute intercollicular decerebration in all experiments resulted in normalization of respiratory rhythm even while ictal ECoG activity continued. We conclude that stimulation of breathing during seizures occurs in the absence of respiratory feedback mechanisms. The findings suggest that an important cause of the respiratory response is a feedforward mechanism, whereby activation of subcortical structures above medulla and pons results in stimulation of breathing.     

Diaphragm EMG measured by cervical magnetic and electrical phrenic nerve stimulation

The purpose of the study was to compare electrical stimulation (ES) and cervical magnetic stimulation (CMS) of the phrenic nerves for the measurement of the diaphragm compound muscle action potential (CMAP) and phrenic nerve conduction time. A specially designed esophageal catheter with three pairs of electrodes was used, with control of electrode positioning in 10 normal subjects. Pair A and pair B were close to the diaphragm (pair A lower than pair B); pair C was positioned 10 cm above the diaphragm to detect the electromyogram from extradiaphragmatic muscles. Electromyograms were also recorded from upper and lower chest wall surface electrodes. The shape of the CMAP measured with CMS (CMS-CMAP) usually differed from that of the CMAP measured with ES (ES-CMAP). Moreover, the latency of the CMS-CMAP from pair B (5.3 +/- 0.4 ms) was significantly shorter than that from pair A (7.1 +/- 0.7 ms). The amplitude of the CMS-CMAP (1.00 +/- 0.15 mV) was much higher than that of ES-CMAP (0.26 +/- 0.15 mV) when recorded from pair C. Good-quality CMS-CMAPs could be recorded in some subjects from an electrode positioned very low in the esophagus. The differences between ES-CMAP and CMS-CMAP recorded either from esophageal or chest wall electrodes make CMS unreliable for the measurement of phrenic nerve conduction time.     

Percutaneous repair of a traumatic bile duct laceration in a liver transplant allograft

BACKGROUND: Phrenic nerve palsy in infants and young children usually results from birth injury or iatrogenic damage. The newborn almost invariably presents with severe respiratory distress, diaphragmatic elevation, and paradoxical movement at the affected side. METHODS/RESULTS: In this retrospective analysis a group of 23 patients below the age of 1 year with an obstetric or postoperative phrenic nerve injury was studied and compared with cases in the literature. All patients were admitted between 1986 and 1997 to the Pediatric Surgical Center, Amsterdam. Thirteen of 18 patients with an obstetric phrenic nerve injury underwent plication of the diaphragm after an average observation period of 100 days. In the remaining five children with an obstetric phrenic nerve injury, spontaneous recovery appeared within 1 month. Only one of five patients with a phrenic nerve palsy after a cardiac surgical procedure underwent plication of the diaphragm. Fifteen of the 34 patients described in the literature underwent plication of the diaphragm after an average of 54 days. CONCLUSIONS: If after 1 month no spontaneous recovery of the diaphragmatic paralysis caused by a phrenic nerve injury occurs, plication of the diaphragm is indicated. This operation proved to be successful for relief of symptomatic phrenic nerve injury in all cases. If the condition of the patient clinically deteriorates during this first month of life, the patient should be operated on immediately.     

Effect of activation of 5-HT1A receptors at the ventral medulla on phrenic nerve activity

The purpose of this study was to determine the effects of the 5-HT1A receptor agonist, 8-hydroxy-2-(di-N-propylamino)tetralin hydrobromide (8-OH-DPAT), on respiratory activity (phrenic nerve activity) following application to the ventral medullary surface in the cat. In addition, in order to determine if the action of 8-OH-DPAT was localized to structures at the ventral medulla, we examined the distribution of [3H]8-OH-DPAT to other brain regions and to the peripheral circulation. 8-OH-DPAT (0.0625-8 micrograms) produced a dose-related increase in respiratory rate when applied at either the intermediate or caudal areas on the ventral surface of the medulla. The maximal change in respiratory rate was 9 +/- 2 and 8 +/- 1 breaths/min at the intermediate and caudal areas, respectively. Integrated phrenic nerve amplitude was not significantly affected at these sites except at the 8 micrograms dose where it was decreased. No change in phrenic nerve activity was observed with 8-OH-DPAT application at the rostral area. [3H]8-OH-DPAT was not found to distribute to other brain regions or to the peripheral circulation following application to the ventral medullary surface. The results of this study suggest that 8-OH-DPAT causes changes in respiratory activity, primarily respiratory rate, by acting on neuronal structures at the ventral surface of the medulla.     

Binding of annexin V to bilayers with various phospholipid compositions using glass beads in a flow cytometer

The effects of intravenous administration of flumazenil (n = 6) or bicuculline (n = 6) on the discharge of the phrenic nerve were studied following vagotomy in pentobarbital anesthetized mechanically ventilated rats. Morphine (0.4 mg.kg-1.min-1) was administrated until the respiratory rate decreased to about a half of the baseline respiratory rate. In this state, we first administered flumazenil (0.25 mg.kg-1) or bicuculline (0.4 mg.kg-1), intravenously and then administered naloxone (0.02 mg) intravenously in the two groups. The increase of inspiratory time from 0.7 +/- 0.1 to 2.0 +/- 0.5 s by morphine recovered to 0.8 +/- 0.2 s by bicuculline and to 0.6 +/- 0.1 s by naloxone. The increase of inspiratory time from 0.7 +/- 0.1 to 1.7 +/- 0.3 s by morphine, and to 2.1 +/- 0.5 s by flumazenil recovered to 0.6 +/- 0.1 s by naloxone. Expiratory time did not change during each drug administration in the two groups. The decrease of respiratory rate from 44 to 23 +/- 4 breaths.min-1 by morphine recovered to 37 +/- 5 breaths.min-1 by bicuculline and to 42 +/- 2 breaths.min-1 by naloxone. The decrease of respiratory rate from 45 +/- 3 to 22 +/- 6 breaths.min-1 by morphine, and to 18 +/- 4 breaths.min-1 by flumazenil recovered to 46 +/- 3 breaths.min-1 by naloxone. Amplitude of integrated phrenic nerve discharge increased to 125 +/- 42% by bicuculline and to 175 +/- 93% by naloxone compared to the baseline values. The decrease of amplitude to 54 +/- 18% by flumazenil recovered to 125 +/- 42% by naloxone. These results suggest that bicuculline not flumazenil antagonizes the respiratory depression of morphine by increasing the respiratory rate and respiratory movement.