Cardiorespiratory Integration Research Articles

Norepinephrine Increases Cytosolic Ca2+ in Neurons of the Paraventricular Nucleus of the Hypothalamus via Multiple Signaling Pathways

The paraventricular nucleus (PVN) of the hypothalamus is a critical cardiorespiratory integration site of central autonomic reflexes. A number of catecholaminergic neurons originating in the nucleus tractus solitarii, ventrolateral medulla, and locus coeruleus project to the PVN and are activated by hypoxia. Within the PVN, norepinephrine (NE) depolarizes neurons and enhances action potential discharge via alpha‐1 adrenergic receptor (α1‐AR) activation. It is unclear what effect, if any, NE has on cytosolic Ca2+. Here we examine the influence of NE on cytosolic Ca2+ in PVN neurons using fura‐2 Ca2+ imaging in isolated PVN cultures. PVN neurons were dissociated from 3–4 week old male Sprague‐Dawley rats and examined within 5 hours of dissociation. Cultures were immunopositive for the neuronal marker PAN and corticotropin‐releasing hormone, a neuropeptide heavily expressed in the PVN, suggesting neuronal identity. Neuronal identity was confirmed by increases in Ca2+ following elevated K+‐induced depolarization. We subsequently monitored the magnitude of basal (non‐phasic) Ca2+ elevation as well the number of responding neurons, defined as those that increased cytosolic Ca2+ by more than one standard deviation above mean vehicle response. NE (100 μM, 5 min, n = 68) induced an increase in basal cytosolic Ca2+ compared to aCSF vehicle (n = 63). This effect was mimicked by the α1‐AR agonist phenylephrine (PE, 10 μM, n = 42) but not the alpha‐2 adrenergic receptor (α2‐AR) agonist clonidine (CLON, 10 μM, n = 28). NE and PE, but not CLON, also significantly increased the number of responding neurons. To examine the mechanism of elevation of basal cytosolic Ca2+ by PE and α1‐AR activation, one or more of its secondary pathways were blocked. Blockade of voltage‐dependent Ca2+ channels with cadmium abolished the Ca2+ increase of PE (n = 60), as did blockers for phospholipase C (PLC, n = 26), protein kinase C (PKC, n = 31), and the inositol trisphosphate receptor (IP3R, n = 53). PKC, PLC and IP3R block reduced the number of responding neurons. Spontaneous (phasic) Ca2+ event frequency and amplitude were not affected by NE, PE, or CLON and were not studied further. However, with repeated phasic K+‐induced membrane depolarization, NE increased baseline Ca2+ yet decreased the depolarization‐induced peak amplitude. PE mimicked the basal increase in Ca2+ without affecting peak amplitude. CLON decreased peak amplitude without affecting basal Ca2+. Taken together, these data suggest NE increases cytosolic Ca2+ in PVN neurons via an α1‐AR mediated pathway which is co‐dependent on extracellular Ca2+ influx and intracellular Ca2+ release, possibly via a PLC‐mediated signaling cascade. This α1‐AR mediated effect does not affect depolarization induced Ca2+ entry, and is not caused by a change in spontaneous Ca2+ event frequency or amplitude.Support or Funding InformationSupport: HL 098602.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Effect of Lower-Body Positive Pressure on the Cardiorespiratory Response at Rest and during Submaximal Running Exercise.

Anti-gravity treadmills facilitate locomotion by lower-body positive pressure (LBPP). Effects on cardiorespiratory regulation are unknown. Healthy men (30 ± 8 y, 178.3 ± 5.7 cm, 70.3 ± 8.0 kg; mean ± SD) stood upright (n = 10) or ran (n = 9) at 9, 11, 13, and 15 km.h−1 (5 min stages) with LBPP (0, 15, 40 mmHg). Cardiac output (CO), stroke volume (SV), heart rate (HR), blood pressure (BP), peripheral resistance (PR), and oxygen uptake (VO2) were monitored continuously. During standing, LBPP increased SV [by +29 ± 13 (+41%) and +42 ± 15 (+60%) ml, at 15 and 40 mmHg, respectively (p < 0.05)] and decreased HR [by −15 ± 6 (−20%) and −22 ± 9 (−29%) bpm (p < 0.05)] resulting in a transitory increase in CO [by +1.6 ± 1.0 (+32%) and +2.0 ± 1.0 (+39%) l.min−1 (p < 0.05)] within the first seconds of LBPP. This was accompanied by a transitory decrease in end-tidal PO2 [by −5 ± 3 (−5%) and −10 ± 4 (−10%) mmHg (p < 0.05)] and increase in VO2 [by +66 ± 53 (+26%) and +116 ± 64 (+46%) ml.min−1 (p < 0.05)], suggesting increased venous return and pulmonary blood flow. The application of LBPP increased baroreflex sensitivity (BRS) [by +1.8 ± 1.6 (+18%) and +4.6 ± 3.7 (+47%) at 15 and 40 mmHg LBPP, respectively P < 0.05]. After reaching steady-state exercise CO vs. VO2 relationships remained linear with similar slope and intercept for each participant (mean R2 = 0.84 ± 0.13) while MAP remained unchanged. It follows that (1) LBPP affects cardiorespiratory integration at the onset of exercise; (2) at a given LBPP, once reaching steady-state exercise, the cardiorespiratory load is reduced proportionally to the lower metabolic demand resulting from the body weight support; (3) the balance between cardiovascular response, oxygen delivery to the exercising muscles and blood pressure regulation is maintained at exercise steady-state; and (4) changes in baroreflex sensitivity may be involved in the regulation of cardiovascular parameters during LBPP.

Prenatal nicotine exposure alters postnatal cardiorespiratory integration in young male but not female rats

The present study tested the hypothesis that prenatal nicotine exposure (PNE) induces sex specific alternations in indices of cardiorespiratory coupling during early development. Rat pups exposed to either nicotine (6mg/kg/day) or saline (control) in utero were chronically instrumented with ECG electrodes for measurement of heart rate (HR) and respiratory frequency (RF) was monitored by whole body plethysmography on postnatal days (P)13, P16 and P26. PNE had no identifiable effect on resting respiratory frequency (RF) in either sex. There was however a strong trend (p=0.057) for resting HR to be elevated by PNE in male offspring only. Alternatively, the HR response to hypoxia (10% O2), was significantly blunted at P13 but significantly elevated at P26 s in the absence of any significant change in RF in PNE males only. Indicators of respiratory sinus arrhythmia (RSA) were also significantly reduced in P26 PNE males. No significant effects of PNE on HR, RF or RSA were identified in female offspring at any age. Our results demonstrate that PNE induces very specific changes in cardiorespiratory integration at select postnatal ages and these changes are more prominent in males. Additionally, alternations in cardiorespiratory integration appear to persist into later development in males only, potentially increasing the risk for cardiovascular diseases such as hypertension later in life.

Breathing disorders in Rett syndrome: Progressive neurochemical dysfunction in the respiratory network after birth

Disorders of respiratory control are a prominent feature of Rett syndrome (RTT), a severely debilitating condition caused by mutations in the gene encoding methyl-CpG-binding protein 2 (MECP2). RTT patients present with a complex respiratory phenotype that can include periods of hyperventilation, apnea, breath holds terminated by Valsalva maneuvers, forced and deep breathing and apneustic breathing, as well as abnormalities of heart rate control and cardiorespiratory integration. Recent studies of mouse models of RTT have begun to shed light on neurologic deficits that likely contribute to respiratory dysfunction including, in particular, defects in neurochemical signaling resulting from abnormal patterns of neurotransmitter and neuromodulator expression. The authors hypothesize that breathing dysregulation in RTT results from disturbances in mechanisms that modulate the respiratory rhythm, acting either alone or in combination with more subtle disturbances in rhythm and pattern generation. This article reviews the evidence underlying this hypothesis as well as recent efforts to translate our emerging understanding of neurochemical defects in mouse models of RTT into preclinical trials of potential treatments for respiratory dysfunction in this disease.

FMRI signal changes in response to forced expiratory loading in congenital central hypoventilation syndrome.

Congenital central hypoventilation syndrome (CCHS) patients show impaired ventilatory responses to CO2 and hypoxia and reduced drive to breathe during sleep but retain appropriate breathing patterns in response to volition or increased exercise. Breath-by-breath influences on heart rate are also deficient. Using functional magnetic resonance imaging techniques, we examined responses over the brain to voluntary forced expiratory loading, a task that CCHS patients can perform but that results in impaired rapid heart rate variation patterns normally associated with the loading challenge. Increased signals emerged in control (n = 14) over CCHS (n = 13; ventilator dependent during sleep but not waking) subjects in the cingulate and right parietal cortex, cerebellar cortex and fastigial nucleus, and basal ganglia, whereas anterior cerebellar cortical sites and deep nuclei, dorsal midbrain, and dorsal pons showed increased signals in the patient group. The dorsal and ventral medulla showed delayed responses in CCHS patients. Primary motor and sensory areas bordering the central sulcus showed comparable responses in both groups. The delayed responses in medullary sensory and output regions and the aberrant reactions in cerebellar and pontine sensorimotor coordination areas suggest that rapid cardiorespiratory integration deficits in CCHS may stem from defects in these sites. Additional autonomic and perceptual motor deficits may derive from cingulate and parietal cortex aberrations.

Quimiorreceptores centrales y síndrome de muerte súbita del lactante

Carbon dioxide partial pressure and pH in the extracellular compartment are the most powerful signals regulating respiration. Central chemoreceptors (QC) undergo the stimulating effect of CO2 and pH upon respiration. This review tries to provide an actual envision of the progress in the knowledgement on central chemoreception. It also tries to highlight the importance of the alterations in the chemoreception mechanism as a cause of sudden infant death syndrome (SIDS). Central chemoreceptors respond to acid-base imbalance acting on neurons that give rise to the central breathing pattern and have the ability to change the respiratory rate, which is normally needed to restore the normal values of acid-base status. QC are widely distributed in the brain stem, however QC neurons in the ventral surface of the medulla like to be the main relays for ventilatory responses after acid-base stimulation. It has been shown that QC are sensitive to pH and CO2 as in vivo as in vitro conditions, most of them being serotonin immunopositive, a neurotransmitter with known effects on breathing pattern. By other side, alterations of central chemoreception have been associated to pathologies like congenital hypoventilatory syndrome or SIDS. Interestingly, the post mortem exams of the brain of infants dead because these syndromes have showed anomalies of serotonergic receptor located in regions containing QC neurons and in others related with cardiorespiratory integration.

Development of hypoxia-induced Fos expression in rat caudal hypothalamic neurons

The caudal hypothalamus is an important CNS site controlling cardiorespiratory integration during systemic hypoxia. Previous findings from this laboratory have identified caudal hypothalamic neurons of anesthetized rats that are stimulated during hypoxia. In addition, patch-clamp recordings in an in vitro brain slice preparation have revealed that there is an age-dependent response to hypoxia in caudal hypothalamic neurons. The present study utilized the expression of the transcription factor Fos as an indicator of neuronal depolarization to determine the hypoxic response of caudal hypothalamic neurons throughout postnatal development in conscious rats. Sprague–Dawley rats, aged three to 56 days, were placed in a normobaric chamber circulated with either 10% oxygen or room air for 3 h. Following the hypoxic/normoxic exposure period, tissues from the caudal hypothalamus, periaqueductal gray, rostral ventrolateral medulla and nucleus tractus solitarius were processed immunocytochemically for the presence of the Fos protein. There was a significant increase in the density of neurons expressing Fos in the caudal hypothalamus of hypoxic compared to normoxic adult rats that was maintained in the absence of peripheral chemoreceptors. In contrast, no increase in the density of Fos-expressing caudal hypothalamic neurons was observed during hypoxia in rats less than 12 days old. Increases in Fos expression were also observed in an age-dependent manner in the periaqueductal gray, rostral ventrolateral medulla and nucleus tractus solitarius. These results show an increase in Fos expression in caudal hypothalamic neurons during hypoxia in conscious rats throughout development, supporting the earlier in vitro reports suggesting that these neurons are stimulated by hypoxia.

Cardiorespiratory Integration in Vertebrates

Cardiorespiratory Integration in Vertebrates

Central control of the cardiovascular and respiratory systems and their interactions in vertebrates.

This review explores the fundamental neuranatomical and functional bases for integration of the respiratory and cardiovascular systems in vertebrates and traces their evolution through the vertebrate groups, from primarily water-breathing fish and larval amphibians to facultative air-breathers such as lungfish and some adult amphibians and finally obligate air-breathers among the reptiles, birds, and mammals. A comparative account of respiratory rhythm generation leads to consideration of the changing roles in cardiorespiratory integration for central and peripheral chemoreceptors and mechanoreceptors and their central projections. We review evidence of a developing role in the control of cardiorespiratory interactions for the partial relocation from the dorsal motor nucleus of the vagus into the nucleus ambiguus of vagal preganglionic neurons, and in particular those innervating the heart, and for the existence of a functional topography of specific groups of sympathetic preganglionic neurons in the spinal cord. Finally, we consider the mechanisms generating temporal modulation of heart rate, vasomotor tone, and control of the airways in mammals; cardiorespiratory synchrony in fish; and integration of the cardiorespiratory system during intermittent breathing in amphibians, reptiles, and diving birds. Concluding comments suggest areas for further productive research.

Synaptic interactions of substance P immunoreactive nerve terminals in the baro- and chemoreceptor reflexes of the cat

The neurochemical anatomy and synaptic interactions of morphologically identified chemoreceptor or baroreceptor afferents in the nucleus of the solitary tract (NTS) are poorly understood. A substantial body of physiological and light microscopic evidence suggests that substance P (SP) may be a neurotransmitter contained in first order sensory chemo- or baroreceptor afferents, however ultrastructural support for this hypothesis is lacking. In the present report we have traced the central projections of the carotid sinus nerve (CSN) in the cat by utilizing the transganglionic transport of horseradish peroxidase. Medullary tissues including the commissural NTS (cNTS) were processed for the histochemical visualization of transganglionically labeled CSN afferents and for the immunocytochemical detection of SP by dual labeling light and electron microscopic methods. At the light microscopic level, dense bilateral labeling with TMB was found in the tractus solitarius (TS) and cNTS, caudal to the obex. Rostral to the obex, significant ipsilateral TMB labeling was detected in the dorsal, dorso-lateral, and medial subnuclei of the NTS, as well as in the TS. Significant staining of SP immunoreactive processes was detected in most subnuclei of the NTS. The cNTS was examined by electron microscopy. Either HRP or SP were readily identified in single labeled unmyelinated axons, myelinated axons, and nerve terminals in the cNTS. SP immunoreactivity was also identified in unmyelinated axons, myelinated axons, and nerve terminals in the cNTS which were simultaneously identified as CSN primary afferents. These ultrastructural data support the hypothesis that SP immunoreactive first order neurons are involved in the origination of the chemo- and baroreceptor reflexes. Axo-axonic synapses were observed between CSN primary afferent terminals and: (a) unlabeled nerve terminals; (b) other CSN primary afferent terminals; and (c) terminals containing SP. Axo-axonic synapses were also observed between CSN primary afferents which contained SP, and other SP terminals. These observations may mediate the morphological bases for multiple forms of presynaptic inhibition in the cNTS, including those involved in cardiorespiratory integration. In conclusion, our results indicate that SP immunoreactive nerve terminals may be important in both the origination and the modulation of the chemo- and/or baroreceptor reflexes.

Periodicities in efferent discharge of splanchnic nerve of the cat.

Periodicities in efferent discharge of splanchnic nerve of the cat.

Ranges of heart rates and their regulations at various ages (rat).

Ranges of heart rates and their regulations at various ages (rat).