Tonic Elevations Research Articles

Generation, breakdown and responses of ATP in the carotid body

Purinergic signaling involving adenosine triphosphate (ATP) acting on P2X2/3 receptors modulates physiological carotid body (CB) afferent discharge and chemoreflex activation however, the upregulation of P2X3 receptors on petrosal sensory neurons is partly responsible for aberrant CB tonicity and hyperreflexia, and the increase in blood pressure in the Spontaneously Hypertensive rat (SHR). The enhanced chemoreflex occurs prior to the onset of hypertension and elevated CB activity is thought to contribute to the underlying pathophysiology of the disease but the exact mechanisms remain unclear. We hypothesize that there is greater release of ATP and/or decreased ATP breakdown in the pre‐hypertensive SHR CB compared with that in the Wistar rat, and this evokes an increase in the chemoreflex response and subsequent activation of the sympathetic nervous system. We performed three studies. (i) Using digital droplet PCR (ddPCR), we measured mRNA expression levels of enzymes involved in the breakdown of ATP in CBs extracted from male 4‐week‐old prehypertensive SHR and Wistar rats; these included: ectonucleotide pyrophosphatase/phosphodiesterase 1‐3 (Enpp1‐3), ectonucleoside triphosphate diphosphohydrolase 2‐3 (Entpd2‐3), and ecto‐5’‐nucleotidase (Nt5e). (ii) We quantified ATP release from CBs in vitro during baseline conditions and during cyanide‐evoked ATP release using a colorimetric assay (213‐579‐1 ‐ Millipore), comparing between male 4‐week‐old pre‐hypertensive SHR and Wistar rats. (iii) In an in situ working heart‐brainstem preparation of 4‐week‐old Wistar rats, simultaneous recordings were made from phrenic (PN), hypoglossal (HN), recurrent laryngeal (RLN), abdominal (ABN), and thoracic sympathetic nerves (tSNA). The peripheral chemoreceptors were stimulated with either potassium cyanide (KCN 20‐100 μL, 0.04%) or ATP at varying concentrations (10‐100 µL, 50 µM ‐ 2 mM) delivered via the internal carotid artery. In whole CB extracts, ddPCR revealed increased Enpp2‐3 and Nt5e expression (p<0.05) in pre‐hypertensive SHRs relative to Wistar rats suggesting greater capacity to break down ATP in the pre‐hypertensive SHR CB. There were no differences in the expressions of Enpp1 or Entpd2‐3 between rat strains. However, the amount of ATP released from the CB in both baseline and during cyanide stimulation was two‐fold greater in the pre‐hypertensive SHR (P<0.05). Stimulation of the CB with KCN evoked hyperpnoea, bradycardia and sympathoexcitation at all doses. In contrast, CB injection of low dose ATP increased PN rate with little effect on tSNA, whereas higher doses induced apnea accompanied by repetitive burst discharge of post‐inspiratory activity in the RLN with tonic elevations in the discharge of both ABN and tSNA. In sum, despite the upregulation of several ATP degrading enzymes in the CBs of the SHR, CB release of ATP is higher in this species. We hypothesize that the increase in ATP release in the SHR CB drives the CB hyperexcitability and that the shift towards an upregulation of ATP degrading enzymes even prior to the development of hypertension is compensatory to oppose the increase in ATP signaling. The dose‐dependent effect of exogenous ATP at supraphysiological levels evoking greater CB‐mediated expiratory and sympathetic outflow in the Wistar rat mimics aspects of pathophysiological CB chemoreflex signaling in the SHR.

Enhanced carotid body chemoreflex function drives respiratory‐sympathetic coupling in systolic heart failure

Enhanced carotid body (CB) chemoreflex sensitivity in systolic heart failure (HF) has been linked to oscillatory breathing patterns and autonomic imbalance. Importantly, breathing pattern alterations and heightened chemoreflex‐mediated sympathetic nerve activity are both associated with higher morbidity and mortality in HF. Therefore, the objective of this study was to determine whether CB denervation (CBD) would improve respiratory stability and decrease sympathetic drive in HF. In rabbits with pacing‐induced HF, we measured resting breathing and renal sympathetic nerve activity (RSNA), before and after CBD. In addition, we assessed respiratory‐sympathetic coupling using power spectral analysis and the magnitude of the peak coherence function between tidal volume (Vt) and RSNA frequency spectra. Minute ventilation and RSNA responses to acute hypoxia were significantly augmented in HF compared to controls and were abolished by CBD. Resting RSNA was greater in HF compared to control animals (43 ± 5% max vs. 23 ± % max, p<0.05). HF animals after CBD displayed resting RSNA levels similar to control animals. HF animals displayed altered breathing patterns characterized by oscillatory breathing and increases in the incidence of apneas/hypopneas. Remarkably, CBD totally normalized oscillatory breathing patterns in HF animals. Peak coherence between RSNA and Vt signals was increased in HF animals (control 0.49 ± 0.05; HF 0.79 ± 0.06), and was significantly attenuated by CBD (0.59 ± 0.05, p<0.05 for all comparisons). Our data show that enhanced CB‐mediated chemoreflex drive contributes to tonic elevations in RSNA and breathing instability in HF. Additionally, a tonic increase in CB afferent discharge may also play an important role in strengthening a link between respiratory and sympathetic neural outflow in HF.Support or Funding InformationSupported by Fondecyt 1140275, NIH PO1‐HL62222 and NIH F32 HL108592‐01A1.

Central role of carotid body chemoreceptors in disordered breathing and cardiorenal dysfunction in chronic heart failure.

Oscillatory breathing (OB) patterns are observed in pre-term infants, patients with cardio-renal impairment, and in otherwise healthy humans exposed to high altitude. Enhanced carotid body (CB) chemoreflex sensitivity is common to all of these populations and is thought to contribute to these abnormal patterns by destabilizing the respiratory control system. OB patterns in chronic heart failure (CHF) patients are associated with greater levels of tonic and chemoreflex-evoked sympathetic nerve activity (SNA), which is associated with greater morbidity and poor prognosis. Enhanced chemoreflex drive may contribute to tonic elevations in SNA by strengthening the relationship between respiratory and sympathetic neural outflow. Elimination of CB afferents in experimental models of CHF has been shown to reduce OB, respiratory-sympathetic coupling, and renal SNA, and to improve autonomic balance in the heart. The CB chemoreceptors may play an important role in progression of CHF by contributing to respiratory instability and OB, which in turn further exacerbates tonic and chemoreflex-evoked increases in SNA to the heart and kidney.

Orienting, emotion, and memory: Phasic and tonic variation in heart rate predicts memory for emotional pictures in men

Arousal-related processes associated with heightened heart rate (HR) predict memory enhancement, especially for emotionally arousing stimuli. In addition, phasic HR deceleration reflects “orienting” and sensory receptivity during perception of stimuli. We hypothesized that both tonic elevations in HR as well as phasic HR deceleration during viewing of pictures would be associated with deeper encoding and better subsequent memory for stimuli. Emotional pictures are more memorable and cause greater HR deceleration than neutral pictures. Thus, we predicted that the relations between cardiac activity and memory enhancement would be most pronounced for emotionally-laden compared to neutral pictures. We measured HR in 53 males during viewing of unpleasant, neutral, and pleasant pictures, and tested memory for the pictures two days later. Phasic HR deceleration during viewing of individual pictures was greater for subsequently remembered than forgotten pictures across all three emotion categories. Elevated mean HR across the entire encoding epoch also predicted better memory performance, but only for emotionally arousing pictures. Elevated mean HR and phasic HR deceleration were associated, such that individuals with greater tonic HR also showed greater HR decelerations during picture viewing, but only for emotionally arousing pictures. Results suggest that tonic elevations in HR are associated both with greater orienting and heightened memory for emotionally arousing stimuli.

Muscle metaboreflex contribution to resting limb haemodynamic control is preserved in older subjects

Ageing is associated with tonic elevations in basal sympathetic vasoconstrictor outflow to skeletal muscle and a parallel decline in vascular function. The purpose of this study was to test the hypothesis that older individuals exhibit attenuated calf vascular resistance (CVR) responses to muscle metaboreflex activation in comparison with young subjects. Fourteen young (mean +/- SD age 23 +/- 3 years) and 13 older (62 +/- 7 years) sedentary subjects participated in the study. To evaluate muscle metaboreflex, we measured heart rate, mean blood pressure (MBP), calf blood flow (CBF) (venous occlusion plethysmography) and CVR responses to static handgrip exercise at 30% of maximal voluntary contraction, followed by recovery with [postexercise circulatory occlusion, (PECO+)] or without (PECO-) circulatory occlusion. Mean BP and CVR increased significantly (ANOVA P<0.05) throughout exercise and remained elevated during PECO+ when compared with PECO- in both groups. There were no significant differences between the two groups in BP and CVR relative changes from baseline during the entire protocol in both trials. CBF responses were also similar in the young and older subjects, except for the first minute of exercise, where young subjects had higher CBF responses. Our results demonstrate that older subjects have similar BP and calf haemodynamic responses to static handgrip exercise and selective action of the muscle metaboreflex when compared with young subjects, compatible with preserved muscle metaboreflex contribution to resting limb haemodynamic control with ageing in humans.

Explaining employees' health care costs: a prospective examination of stressful job demands, personal control, and physiological reactivity.

The authors tested the ability of stressful demands and personal control in the workplace to predict employees' subsequent health care costs in a sample of 105 full-time nurses. Both subjective and objective measures of workload demands interacted with personal control perceptions in predicting the cumulative health care costs over the ensuing 5-year period. Tonic elevations in salivary cortisol, moreover, mediated the effects of demands and control on health care costs. Neither the job demands variables nor physiological reactivity measures, however, explained subsequent mental health. The results support findings from the epidemiological literature that demonstrate an important role for employees' control in explaining occupational inequalities in coronary heart disease and mortality. The authors argue that the results also encourage control-enhancing job design interventions by suggesting that their outcomes can benefit both organizations and their members.

Calcium imaging of rhythmic network activity in the developing spinal cord of the chick embryo

Video-rate imaging of spinal neurons loaded with calcium-sensitive dyes was used to investigate the calcium dynamics and cellular organization of spontaneously active rhythm-generating networks in the spinal cord of E9-E12 chick embryos. Spinal neurons were loaded with bath-applied fura-2am. Motoneurons were also loaded by retrograde labeling with dextran-conjugated, calcium-sensitive dyes. Dye-filled motoneurons exhibited large fluorescent changes during antidromic stimulation of motor nerves, and an increase in the 340/380 fura fluorescence ratio that is indicative of increased intracellular free calcium. Rhythmic fluorescence changes in phase with motoneuron electrical activity were recorded from motoneurons and interneurons during episodes of evoked or spontaneous rhythmic motor activity. Fluorescent responses were present in the cytosol and in the perinuclear region, during antidromic stimulation and network-driven rhythmic activity. Optically active cells were mapped during rhythmic activity, revealing a widespread distribution in the transverse and horizontal planes of the spinal cord with the highest proportion in the ventrolateral part of the cord. Fluorescent signals were synchronized in different regions of the cord and were similar in time course in the lateral motor column and in the intermediate region. In the dorsal region the rhythm was less pronounced and the signal decayed after a large initial transient. Video-rate fluorescent measurements from individual cells confirmed that fluorescent signals were synchronized in interneurons and in motoneurons although the time course of the signal could vary between cells. Some of the interneurons exhibited tonic elevations of fluorescence for the duration of the episode whereas others were rhythmically active in phase with motoneurons. At the onset of each cycle of rhythmic activity the earliest fluorescent change occurred ventrolaterally, in and around the lateral motor column, from which it spread to the rest of the cord. The results suggest that neurons in the ventrolateral part of the spinal cord are important for rhythmogenesis and that axons traveling in the ventrolateral white matter may be involved in the rhythmic excitation of motoneurons and interneurons. The widespread synchrony of the rhythmic calcium transients may reflect the existence of extensive excitatory interconnections between spinal neurons. The network-driven calcium elevations in the cytosol and the perinuclear region may be important in mediating activity-dependent effects on the development of spinal neurons and networks.

In vivo estimates of extracellular dopamine and dopamine metabolite levels during intravenous cocaine or heroin self-administration

Intravenous opiate and stimulant self-administration are characterized by regularly spaced responses and relatively constant hourly drug intake. Recent in vivo neurochemical studies have revealed tonic elevations in extracellular dopamine levels in nucleus accumbens—a transmitter and a site implicated in both opiate and stimulant reward—that are correlated with the tonic elevations of heroin or cocaine in the blood of animals engaged in drug self-administration. Current studies focus on phasic neurochemical fluctuations—time-locked to the cycle of falling and rising drug levels—which correlate with within-session phases of drug-seeking and drug-satiety, respectively. Improvements in the temporal and neurochemical resolution of current methods are needed before the brain correlates of drug craving and drug reward can be fully characterized.

Activity of cat locus coeruleus noradrenergic neurons during the defense reaction

The single-unit activity of locus coeruleus noradrenergic (LC-NE) neurons was recorded in freely moving cats during naturally induced defense reactions. Defense reactions, consisting of arched back, piloerection, flattened ears and mydriasis, were elicited by exposing the cat either to a dog, or to a cat displaying aggressive behavior induced by electrical stimulation of the hypothalamus. LC-NE neurons were identified using previously established criteria, including suppression of firing during rapid eye movement (REM) sleep and in response to clonidine administration. Exposure to a dog evoked defense reactions and increased the tonic firing rate of LC-NE neurons ( n = 8) from a baseline of ∼0.9 spikes/s to ∼2.5 spikes/s. Exposure to an aggressive cat evoked defense reactions that were qualitatively very similar to those produced by the dog exposure, and elevated the tonic firing rate of LC-NE neurons ( n = 8) from a baseline of ∼1.0 spikes/s to ∼2.5 spikes/s. In addition to these tonic elevations of activity, LC-NE neurons discharged in phasic bursts (as high as 10 spikes in a 500 ms period) in close association with specific threatening acts made by the dog or hypothalamically stimulated cat. The mere presence of a dog was sufficient to evoke tonic activation of LC-NE neurons, even in the absence of threatening advances by the dog, whereas exposure to a hypothalamically stimulated cat produced LC-NE neuronal activation only when the stimulated cat showed aggressive behavior. These results extend our previous work, which examined the response of LC-NE neurons to environmental and physiological stressors, into a more ethologically relevant domain, and suggest that LC-NE neuronal activation may play a role in the response to threatening or challenging situations.

Single-unit response of noradrenergic neurons in the locus coeruleus of freely moving cats. II. Adaptation to chronically presented stressful stimuli

The preceding paper described a stress-related tonic activation of noradrenergic neurons of the locus coeruleus (LC) of behaving cats. The present study examined the degree to which adaptation to stress was reflected in LC unit activity. Adaptation was defined as a reduction in the magnitude of the physiological stress response of the animal, as reflected in plasma norepinephrine level and heart rate. Adaptation of the physiological stress response occurred after 5 repeated hourly presentations of 100 dB white noise and after 2 hr of restraint. When these stimuli ceased to elicit significant sympathoadrenal activation, they likewise ceased to elicit activation of LC single-unit activity. These results provide further support for the hypothesis that tonic elevations in LC neuronal activity are stress related and that the LC is involved in the CNS response to challenges to the organism.

Gastrointestinal motility disturbances in patients with orthostatic hypotension

Orthostatic hypotension syndromes may be associated with visceral denervation, resulting in disturbances of the gastrointestinal and urinary systems. This report analyzes the findings in 9 patients with neurogenic orthostatic hypotension and gastrointestinal symptoms. Antral and proximal small intestinal motor dysfunction was demonstrated in all patients: a significant reduction (p = 0.007) in the number of interdigestive motor complexes during 3 h of fasting (mean 0.4 vs. 1.3 controls); absence of gastric component in interdigestive motor complexes observed; incoordination of fasting and fed antral activity; and development of nonpropagating bursts of phase III-like activity in the intestine that were of at least 2-min duration, high amplitude and frequency, and associated with tonic elevations in baseline pressure. These motor abnormalities may provide a potential means for the diagnosis of impaired autonomic supply to the upper gut.

Myoelectric Activity and Intraluminal Pressure of the Canine Ileocolonic Sphincter

We constructed ileocolonic loops of distal ileum, ileocolonic sphincter, and proximal colon in 5 dogs; neuromuscular continuity was maintained between loop and intact bowel by a bridge of tunica muscularis. To study ileocolonic sphincter function, we recorded myoelectric activity from subserosal electrodes and intraluminal pressures from perfused catheters and a sleeve sensor. Recordings included at least one interdigestive myoelectric complex and four postprandial hours. Ileal slow waves were recorded from the loop and the ileocolonic sphincter; most interdigestive myoelectric complexes were propagated to the ileocolonic sphincter. Intraluminal pressures at the ileocolonic sphincter fluctuated phasically (mean 12.5 cycles/min) for 72% of recording time during fasting and 81% postprandially. Tonic pressures at the ileocolonic sphincter were higher during fasting (31 +/- 18 cmH2O; p less than 0.05) than postprandially (24 +/- 20 cmH2O); moreover, tonic pressures fluctuated more during fasting than after food. Tonic elevations of pressure were prolonged (mean duration 28.5 min) concomitant with arrival of phase 3 of interdigestive myoelectric complexes at the ileocolonic sphincter. Thus the ileocolonic sphincter participates in both the myoelectric and motor components of the interdigestive cycle and exhibits an intense phasic and tonic response to phase 3. The association of tonic pressure and intense phasic contractions may serve to retard and segment ileal chyme. On the other hand, the presence of rapidly propagated phasic pressures in the region may provide the basis for propulsive function. In this way, these unique motor events will facilitate two distinct functions of the region, namely the alternate retarding and forward propulsion of chyme.

The effects of conflict on tonic levels of blood pressure in the genetically borderline hypertensive rat.

ABSTRACTStudies of aversive conditioning in animals have seldom elicited tonic elevations in blood pressure (BP) equivalent to those which produce pathological changes in humans. While it is possible that psychological factors are not that important in the etiology of hypertension, it is also possible that the failure to elicit large tonic elevations in BP in animals may be due either to stressors which are insufficiently potent and/or to an inadequate physiological model. The present study sought to maximize the probability of producing large tonic changes in BPs by using a conflict paradignt in a genetic strain of rats which develops systolic BPs in the borderline hypertensive area (c. 150 mmHg). Forty‐eight male F1 generation offspring of spontaneously hypertensive rats mated with normotensive controls were randomly split into three groups: experimental (subjected to 3 weeks of avoidance training and 12 weeks of conflict in conditioning cages), mild restraint control (placed in conditioning cages daily but not shocked), and maturation control (neither shocked nor restrained) groups. Animals subjected to conflict gradually developed tonic levels of systolic BP well into the hypertensive range (c. 185 mmHg). Restraint control animals also showed some elevation (c. 165 mmHg), but maturational controls showed no change (c. 150 mmHg). The saliency of this animal model for the study of stress‐induced hypertension is discussed.