Cyclical modulation of human ventricular repolarization by respiration.

BACKGROUND: Respiratory modulation of autonomic input to the sinus node results in cyclical modulation of heart rate, known as respiratory sinus arrhythmia (RSA). We hypothesized that the respiratory cycle may also exert cyclical modulation on ventricular repolarization, which may be separately measurable using local endocardial recordings. METHODS AND RESULTS: The study included 16 subjects with normal ventricles undergoing routine clinical electrophysiological procedures for supraventricular arrhythmias. Unipolar electrograms were recorded from 10 right and 10 left ventricular endocardial sites. Breathing was voluntarily regulated at 5 fixed frequencies (6, 9, 12, 15, and 30 breaths per min) and heart rate was clamped by RV pacing. Activation-recovery intervals (ARI: a surrogate for APD) exhibited significant (p < 0.025) cyclical variation at the respiratory frequency in all subjects; ARI shortened with inspiration and lengthened with expiration. Peak-to-peak ARI variation ranged from 0-26 ms; the spatial pattern varied with subject. Arterial blood pressure also oscillated at the respiratory frequency (p < 0.025) and lagged behind respiration by between 1.5 s and 0.65 s from slowest to fastest breathing rates respectively. Systolic oscillation amplitude was significantly greater than diastolic (14 ± 5 vs. 8 ± 4 mm Hg ± SD, p < 0.001). CONCLUSIONS: Observations in humans with healthy ventricles using multiple left and right ventricular endocardial recordings showed that ARI action potential duration (APD) varied cyclically with respiration.

Cardioprotection evoked by remote ischaemic preconditioning is critically dependent on the activity of vagal pre-ganglionic neurones.

AIMS: Innate mechanisms of inter-organ protection underlie the phenomenon of remote ischaemic preconditioning (RPc) in which episode(s) of ischaemia and reperfusion in tissues remote from the heart reduce myocardial ischaemia/reperfusion injury. The uncertainty surrounding the mechanism(s) underlying RPc centres on whether humoral factor(s) produced during ischaemia/reperfusion of remote tissue and released into the systemic circulation mediate RPc, or whether a neural signal is required. While these two hypotheses may not be incompatible, one approach to clarify the potential role of a neural pathway requires targeted disruption or activation of discrete central nervous substrate(s). METHODS AND RESULTS: Using a rat model of myocardial ischaemia/reperfusion injury in combination with viral gene transfer, pharmaco-, and optogenetics, we tested the hypothesis that RPc cardioprotection depends on the activity of vagal pre-ganglionic neurones and consequently an intact parasympathetic drive. For cell-specific silencing or activation, neurones of the brainstem dorsal motor nucleus of the vagus nerve (DVMN) were targeted using viral vectors to express a Drosophila allatostatin receptor (AlstR) or light-sensitive fast channelrhodopsin variant (ChIEF), respectively. RPc cardioprotection, elicited by ischaemia/reperfusion of the limbs, was abolished when DVMN neurones transduced to express AlstR were silenced by selective ligand allatostatin or in conditions of systemic muscarinic receptor blockade with atropine. In the absence of remote ischaemia/reperfusion, optogenetic activation of DVMN neurones transduced to express ChIEF reduced infarct size, mimicking the effect of RPc. CONCLUSION: These data indicate a crucial dependence of RPc cardioprotection against ischaemia/reperfusion injury upon the activity of a distinct population of vagal pre-ganglionic neurones.

Impact of lung inflation cycle frequency on rat muscle and skin sympathetic activity recorded using suction electrodes.

Microneurography has been used in humans to study sympathetic activity supplying targets within skeletal muscle and skin. Comparable animal studies are relatively few, probably due to the technical demands of traditional fibre picking techniques. Here we apply a simple suction electrode technique to record cutaneous (CVC) and muscle (MVC) vasoconstrictor activities and describe and investigate the basis of the frequency dependence of lung inflation related modulation. Hindlimb MVC and CVC activities were recorded concurrently. The magnitude of MVC and CVC activities at the lung inflation cycle frequency was significantly less at 2.0 Hz than at lung inflation cycle frequencies < or =1.0 Hz. As lung inflation cycle frequency was increased the coherence between lung inflation cycle or BP and MVC or CVC waveforms decreased. Consistent with the hypothesis that much of the coherence between lung inflation cycle and nerve activity waveforms is secondary to oscillating baroreceptor activity attributable to BP waves, partialization with the BP waveform significantly decreased the coherence between lung inflation cycle and nerve waveforms, and there was an absence of coherence between these waveforms following sinus and aortic denervation. Our data extend findings from other laboratories and establish the value of a suction electrode technique for recording MVC and CVC activities. Furthermore, our observations describe the rates of positive pressure ventilation that avoid strong and regular gating of sympathetic activity.

Immunohistochemical detection of connexin36 in sympathetic preganglionic and somatic motoneurons in the adult rat.

Gap junctional communication in the adult CNS plays an important role in the synchronization of neuronal activities. In vitro studies have shown evidence of electrotonic coupling through gap junctions between sympathetic preganglionic motoneurons and between somatic motoneurons in the neonatal and adult rat spinal cord. Electrotonic transmission of membrane oscillations might be an important mechanism for recruitment of neurons and result in the generation of rhythmic sympathetic and somato-motor activity at the population level. Gap junctions in the adult spinal cord are constituted principally by connexin36 (Cx36). However, the distribution of Cx36 in specific neuronal populations of the spinal cord is unknown. Here, we identify Cx36-like immunoreactivity in sympathetic preganglionic and somatic motoneurons in thoracic spinal cord segments of the adult rat. For this purpose, double immunostaining against Cx36 and choline acetyltransferase (ChAT) was performed on transverse sections (20 microm) taken from spinal segments T6-T8. Cx36 punctate immunostaining was detected in the majority of ChAT-immunoreactive (-ir) neurons from lamina VII [intermediolateral cell column (IML) and intercalated cell group (IC)], lamina X [central autonomic nucleus (CA)] and in ventral horn neurons from laminae VIII and IX. Cx36 puncta were distributed in the neuronal somata and along dendritic processes. The presence of Cx36 in ChAT-ir neurons is consistent with electrical coupling between sympathetic preganglionic motoneurons and between somatic motoneurons through gap junctions in the adult spinal cord.

Sympathetic rhythms and nervous integration.

1. The present review focuses on some of the processes producing rhythms in sympathetic nerves influencing cardiovascular functions and considers their potential relevance to nervous integration. 2. Two mechanisms are considered that may account for rhythmic sympathetic discharges. First, neuronal elements of peripheral or central origin produce rhythmic activity by phasically exciting and/or inhibiting neurons within central sympathetic networks. Second, rhythms arise within central sympathetic networks. Evidence is considered that indicates the operation of both mechanisms; the first in muscle and the second in skin sympathetic vasoconstrictor networks. 3. Sympathetic activity to the rat tail, a model for the nervous control of skin circulation, is regulated by central networks involved in thermoregulation and those associated with fear and arousal. In an anaesthetized preparation, activity displays an apparently autonomous rhythm (T-rhythm; 0.4-1.2 Hz) and the level of activity can be manipulated by regulating core body temperature. This model has been used to study rhythm generation in central sympathetic networks and possible functional relevance. 4. A unique insight provided by the T rhythm, into possible physiological function(s) underlying rhythmic sympathetic discharges is that the activity of single sympathetic post-ganglionic neurons within a population innervating the same target can have different rhythm frequencies. Therefore, the graded and dynamic entrainment of the rhythms by inputs, such as central respiratory drive and/or lung inflation-related afferent activity, can produce graded and dynamic synchronization of sympathetic discharges. The degree of synchronization may influence the efficacy of transmission in a target chain of excitable cells. 5. The T-rhythm may be generated within the spinal cord because the intrathecal application of 5-hydroxytryptamine at the L1 level of the spinal cord of a rat spinalized at T10-T11 produces a T-like rhythm. Thus, induction and modulation of spinal cord oscillators may be mechanisms that influence ganglionic and neuroeffector transmission. 6. The study of sympathetic rhythms may not only further understanding of sympathetic control, but may also inform on the relevance of rhythmic nervous activities in general.

Generation of a physiological sympathetic motor rhythm in the rat following spinal application of 5-HT.

When applied in vitro to various CNS structures 5-HT and/or NMDA have been observed to generate rhythmic nervous activity. In contrast, reports of similar in vivo actions are relatively rare. Here we describe a physiological sympathetic motor rhythm regulating the thermoregulatory circulation of the rat tail (T-rhythm; 0.40-1.20 Hz) that can be elicited following intrathecal (i.t.) application of 5-HT to an in situ'isolated' spinal cord preparation (anaesthetized rats spinalized at T10-T11 and cauda equina cut). i.t. injections were delivered to L1 as sympathetic neuronal activity to the tail (SNAT) arises from preganglionic neurones at T11-L2. SNAT was abolished after spinal transection (n = 18) and it did not return spontaneously. The administration of 5-HT (250 nmol) generated rhythmic sympathetic discharges (n = 6). The mean frequency of the T-like rhythm during the highest level of activity was 0.88 +/- 0.04 Hz which was not significantly different from the T-rhythm frequency observed in intact animals (0.77 +/- 0.02 Hz; P > 0.05 n = 16). In contrast, NMDA (1 micromol) generated an irregular tonic activity, but it failed to generate a T-like rhythm (n = 9), even though the mean levels of activity were not significantly different to those produced by 5-HT. However, 5-HT (250 nmol) applied after NMDA generated a T-like rhythm (0.95 +/- 0.11 Hz, n = 6). Our observations support the idea that 5-HT released from rostral ventromedial medullary neurones, known to innervate sympathetic preganglionic neurones, can induce sympathetic rhythmic activity.

A comparison of simultaneously recorded muscle and skin vasoconstrictor population activities in the rat using frequency domain analysis.

In anaesthetized rats, an apparently autonomous sympathetic rhythm (T-rhythm, frequency range 0.4-1.2 Hz), has been observed in nerve activity controlling thermoregulatory circulations but not renal nerves. To further explore the differential control of sympathetic activity here, we investigate whether the so-called T-rhythm is a feature of muscle vasoconstrictor (MVC) population activity. Population activity was studied in vagotomised anaesthetised rats (alpha-chloralose or urethane maintenance, after barbiturate or halothane induction, respectively). Some rats were additionally sino-aortic denervated (SAD) and/or given a pneumothorax and neuromuscular blocked. Animals were held in central (hypocapnic) apnoea (ventilated at 2 Hz, tidal volume