Carotid Sinus Nerve Research Articles

Relationship of Baseline NT-proBNP to Responsiveness to Vagus Nerve Stimulation in Patients with Heart Failure and Reduced Ejection Fraction

Background Recent studies of patients with HFrEF have suggested that lower baseline NT-proBNP levels are associated with improved morbidity/mortality outcomes with pharmaceutical treatment. Neuromodulation (NM) using carotid nerve plexus stimulation has also been associated with better clinical outcomes when baseline NT-proBNP Objective We evaluated the relationship of baseline NT-proBNP to changes in symptoms and function in patients during NM using VNS for 6 months in ANTHEM-HF. Methods NT-proBNP was collected for exploratory analysis and not used as an inclusion criterion for the study. A repeated measures, generalized-estimating, equations model evaluated the relationship of baseline NT-proBNP values above and below 1,600 pg/ml to symptomatic and functional responses. Results The median (interquartile range; maximum) NT-proBNP was 868 (322, 1875; 14,656) pg/ml (n=58). Heart rate (HR), HR variability (SDNN), 6-minute walk distance, MLWHF mean score, and NYHA improved significantly at 6 months, independent of baseline NT-proBNP. While there was a statistical interaction observed between lower baseline NT-proBNP and better LVEF improvement, LVEF improved clinically in the overall cohort (32±7 to 37±10% [p=0.0042]), and in the two subgroups with NT-proBNP below or above the median for the study population (36±6 to 42±10% [n=29; p Conclusions In ANTHEM-HF, overall symptomatic and functional response to ART was not associated with baseline NT-proBNP. These findings are preliminary and hypothesis-generating. The ongoing ANTHEM-HFrEF Pivotal Study (NCT03425422) is likely to provide additional insight.

Role of the Carotid Body in an Ovine Model of Renovascular Hypertension.

The carotid body is implicated as an important mediator and potential treatment target for hypertension. The mechanisms driving increased carotid body tonicity in hypertension are incompletely understood. Using a large preclinical animal model, which is crucial for translation, we hypothesized that carotid sinus nerve denervation would chronically decrease blood pressure in a renovascular ovine model of hypertension in which hypertonicity of the carotid body is associated with reduced common carotid artery blood flow. Adult ewes underwent either unilateral renal artery clipping or sham surgery. Two weeks later, flow probes were placed around the contralateral renal and common carotid arteries. Hypertension was accompanied by a significant reduction in common carotid blood flow but no change in renal blood flow. Carotid sinus nerve denervation significantly reduced blood pressure compared with sham. In both hypertensive and normotensive animals, carotid body stimulation using potassium cyanide caused dose-dependent increases in mean arterial pressure and common carotid conductance but a reduction in renal vascular conductance. These responses were not different between the animal groups. Taken together, our findings indicate that (1) the carotid body is activated in renovascular hypertension, and this is associated with reduced blood flow (decreased vascular conductance) in the common carotid artery and (2) the carotid body can differentially regulate blood flow to the common carotid and renal arteries. We suggest that in the ovine renovascular model, carotid body hypertonicity may be a product of reduced common carotid artery blood flow and plays an amplifying role with the kidney in the development of hypertension.

Topographic carotid vasoconstriction in the rostral ventrolateral medulla of rats.

The vascular beds of various cranial tissues receive common carotid flow, which contributes to blood flow redistribution associated with animal behaviors such as grooming, but the medullary autonomic regulation of carotid flow resistance (CAR) is poorly understood. This study is the first to examine the response sites of CAR in the rat rostral ventrolateral medulla (RVLM) presympathetic area to chemical stimulation by the ionotropic excitatory amino acid receptors activator L-cysteine. Arterial blood pressure and CAR were monitored in anesthetized rats which had a cranial window constructed above the ventral medulla. Mapping of L-cysteine microinjection in eight rats showed carotid vasoconstriction in the caudal part alone within the RVLM pressor area, which included contributions from other vascular beds, indicating localized topographic carotid vasoconstriction. Additional testing was performed on four types of denervated rats. A similar response map was obtained in six rats that received minimal lesions during surgery as well as in 10 rats with severed internal or external carotid nerves. However, the remaining three minimally lesioned rats showed extensive vasoconstriction of the RVLM pressor area including the rostral part, indicating lack of a topographic response. The topographic response of most rats might be state-dependent. Seven rats with complete cervical denervation showed no carotid vasoconstrictor response in the RVLM pressor area, indicating cervical sympathetic mediation of the responses. The topographic carotid vasoconstriction in response to L-cysteine may suggest differential roles of presympathetic neurons in the rostral and caudal parts of the RVLM in sympathetic carotid flow regulation.

Fail-safe aspects of oxygen supply.

A fall in oxygen supply releases a remedial response that is otherwise prevented when the oxygen supply is sufficient; for example, the remedial function of HIF-1α is released when oxygen levels fall. the physiological responses initiated when oxygen supply is compromised operate in a fail-safe manner. This concept is applied to two cases: the control of cerebral blood flow, and the detection of hypoxia by the carotid body. The fail-safe oxygen supply concept was tested with simple computer simulations to demonstrate its function and verify the ability to reproduce measured data. The computer model reproduced published observations, suggesting that the fail-safe concept can be considered as a principle that provides novel insight into the physiology of oxygen supply in these cases. An engineered fail-safe system automatically prevents or mitigates the consequences of a system failure. This operational concept can be applied both to the delivery of oxygen to the brain during hypoxia and anaemia, and to the carotid body response to hypoxia and hypercapnia. I aimed to develop simple mathematical models of these fail-safe processes and examine their ability to replicate experimental observations. The intent is to demonstrate the validity of applying the fail-safe concept, not to reveal the details of the physiology involved. The model calculations are based on a single compartment of the relevant tissue in each case that is challenged with a decrease in oxygen supply. The model equation parameters were adjusted to reproduce experimental observations. The fail-safe model of cerebral blood flow control yielded results similar in form to published experimental observations of the cerebral blood flow responses to hypoxia and anaemia. The fail-safe model of carotid body glomus cell control of intracellular hydrogen ion concentration also yielded results similar in form to observations of carotid sinus nerve responses to hypoxia and hypercapnia. The ability of these simple models to simulate experimental observations demonstrates the applicability of the fail-safe concept to oxygen delivery. I suggest that a fail-safe view of oxygen delivery provides novel physiological insight.

The potential role of the carotid body in COVID-19.

The carotid body (CB) plays a contributory role in the pathogenesis of various respiratory, cardiovascular, renal, and metabolic diseases through reflex changes in ventilation and sympathetic output. On the basis of available data about peripheral arterial chemoreception and severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), a potential involvement in the coronavirus disease 2019 (COVID-19) may be hypothesized through different mechanisms. The CB could be a site of SARS-CoV-2 invasion, due to local expression of its receptor [angiotensin-converting enzyme (ACE) 2] and an alternative route of nervous system invasion, through retrograde transport along the carotid sinus nerve. The CB function could be affected by COVID-19-induced inflammatory/immune reactions and/or ACE1/ACE2 imbalance, both at local or systemic level. Increased peripheral arterial chemosensitivity and reflex sympatho-activation may contribute to the increased morbidity and mortality in COVID-19 patients with respiratory, cardiovascular, renal, or metabolic comorbidities.

Exploring the Mediators that Promote Carotid Body Dysfunction in Type 2 Diabetes and Obesity Related Syndromes.

Carotid bodies (CBs) are peripheral chemoreceptors that sense changes in blood O2, CO2, and pH levels. Apart from ventilatory control, these organs are deeply involved in the homeostatic regulation of carbohydrates and lipid metabolism and inflammation. It has been described that CB dysfunction is involved in the genesis of metabolic diseases and that CB overactivation is present in animal models of metabolic disease and in prediabetes patients. Additionally, resection of the CB-sensitive nerve, the carotid sinus nerve (CSN), or CB ablation in animals prevents and reverses diet-induced insulin resistance and glucose intolerance as well as sympathoadrenal overactivity, meaning that the beneficial effects of decreasing CB activity on glucose homeostasis are modulated by target-related efferent sympathetic nerves, through a reflex initiated in the CBs. In agreement with our pre-clinical data, hyperbaric oxygen therapy, which reduces CB activity, improves glucose homeostasis in type 2 diabetes patients. Insulin, leptin, and pro-inflammatory cytokines activate the CB. In this manuscript, we review in a concise manner the putative pathways linking CB chemoreceptor deregulation with the pathogenesis of metabolic diseases and discuss and present new data that highlight the roles of hyperinsulinemia, hyperleptinemia, and chronic inflammation as major factors contributing to CB dysfunction in metabolic disorders.

Hypoxia-induced hypotension elicits adenosine-dependent phrenic long-term facilitation after carotid denervation

Moderate acute intermittent hypoxia (AIH) elicits a persistent, serotonin-dependent increase in phrenic amplitude, known as phrenic long-term facilitation (pLTF). Although pLTF was originally demonstrated by carotid sinus nerve stimulation, AIH still elicits residual pLTF in carotid denervated (CBX) rats via a distinct, but unknown mechanism. We hypothesized that exaggerated hypoxia-induced hypotension after carotid denervation leads to greater spinal tissue hypoxia and extracellular adenosine accumulation, thereby triggering adenosine 2A receptor (A2A)-dependent pLTF. Phrenic activity, arterial pressure and spinal tissue oxygen pressure were measured in anesthetized CBX rats. Exaggerated hypoxia-induced hypotension after CBX was prevented via intravenous phenylephrine; without the hypotension, spinal tissue hypoxia during AIH was normalized, and residual pLTF was no longer observed. Spinal A2A (MSX-3), but not serotonin 2 receptor (5-HT2) inhibition (ketanserin), abolished residual pLTF in CBX rats. Thus, pLTF regulation may be altered in conditions impairing sympathetic activity and arterial pressure regulation, such as spinal cord injury.

Baseline NT-proBNP and responsiveness to autonomic regulation therapy in patients with heart failure and reduced ejection fraction

BackgroundRecent heart failure studies have associated lower baseline natriuretic peptide levels with improved morbidity/mortality outcomes during pharmaceutical treatment, and better clinical outcomes during neuromodulation (NM) with carotid nerve plexus stimulation for HFrEF when NT-proBNP < 1600 pg/ml. Whether baseline NT-proBNP is associated with HFrEF responsiveness to NM using vagus nerve stimulation (VNS) has not been examined. Hence, we evaluated the interaction of baseline NT-proBNP with changes in symptoms and function that occurred during chronic VNS in the ANTHEM-HF study. MethodsA repeated measures, generalized-estimating, equations model evaluated the relationship of baseline NT-proBNP values above and below 1600 pg/ml to symptomatic and functional responses in ANTHEM-HF. ResultsMedian (interquartile range; maximum) NT-proBNP was 868 (322, 1875; 14,656) pg/ml (N = 58). Heart rate (HR), HR variability (SDNN), 6-minute walk distance, MLWHF mean score, and NYHA improved significantly, independent of baseline NT-proBNP. While there was a statistical interaction between baseline NT-proBNP and better LVEF improvement during VNS, LVEF improved overall in the study cohort (N = 60; 32 ± 7 to 37 ± 10%; p = 0.0042), and in those patients whose baseline NT-proBNP was below the median baseline NT-proBNP value (n = 29; 36 ± 6 to 42 ± 10%; p < 0.0025)] or above this value (n = 29; 29 ± 7 to 32 ± 9%; p < 0.05). ConclusionsIn ANTHEM-HF, overall symptomatic and functional improvement during chronic VNS was independent of baseline NTproBNP. These are preliminary and hypothesis-generating findings, and the reason for a differing interaction between baseline NT-proBNP and response to CNPS and VNS remains unclear. It is anticipated that the ongoing ANTHEM-HFrEF Pivotal Study of VNS will provide additional insight.

Plasticity of cardiovascular chemoreflexes after prolonged unilateral carotid body denervation: implications for its therapeutic use.

Unilateral carotid body denervation has been proposed as treatment for sympathetic-related human diseases such as systolic heart failure, hypertension, obstructive sleep apnea, and cardiometabolic diseases. The long-term therapeutic effects of carotid body removal will be maintained if the remnant "buffer nerves," that is, the contralateral carotid nerve and the aortic nerves that innervate second-order neurons at the solitary tract nuclei (NTS), do not modify their contributions to the cardiovascular chemoreflexes. Here, we studied the cardiovascular chemoreflexes 1 mo after unilateral carotid body denervation either by excision of the petrosal ganglion (petrosal ganglionectomy, which eliminates central carotid afferents) or exeresis of a segment of one carotid nerve (carotid neurectomy, which preserves central afferents). Cardiovascular chemoreflexes were induced by intravenous (iv) injections of sodium cyanide in pentobarbitone-anesthetized adult cats. After 1 mo of unilateral petrosal ganglionectomy, without significant changes in basal arterial pressure, the contribution of the contralateral carotid nerve to the chemoreflex increases in arterial pressure was enhanced without changes in the contribution provided by the aortic nerves. By contrast, after 1 mo of unilateral carotid neurectomy, the contribution of remnant buffer nerves to cardiovascular chemoreflexes remained unmodified. These results indicate that a carotid nerve interruption involving denervation of second-order chemosensory neurons at the NTS will trigger cardiovascular chemoreflex plasticity on the contralateral carotid pathway. Then, unilateral carotid body denervation as therapeutic tool should consider the maintenance of the integrity of carotid central chemoafferents to prevent plasticity on remnant buffer nerves.NEW & NOTEWORTHY Unilateral carotid body denervation has been proposed as treatment for sympathetic hyperactivity-related human disorders. Its therapeutic effectiveness for maintaining a persistent decrease in the sympathetic outflow activity will depend on the absence of compensatory chemoreflex plasticity in the remnant carotid and aortic afferents. Here, we suggest that the integrity of central afferents after carotid body denervation is essential to prevent the emergence of plastic functional changes on the contralateral "intact" carotid nerve.

Effect of Superior Cervical Gangionectomy on the Chemoreflex Response to Hypoxia in the Rat Carotid Body

Carotid body (CB) chemosensitive glomus (type I) cells respond to decreases in blood pO2 (hypoxia) by releasing neurotransmitters, which in turn activate adjacent chemosensory afferent fibers within the carotid sinus nerve (CSN). These CSN chemoafferents then signal to the commissural nucleus tractus solitarius (cNTS) within the dorsal brainstem, which leads to a compensatory increase in ventilation in order to restore blood‐gas homeostasis. The CB receives sympathetic innervation via the ganglioglomerular nerve (GGN), whose cell bodies reside in the superior cervical ganglion (SCG). We have found that electrical stimulation of the GGN causes an increase in CB chemosensory afferent discharge. However, it is unknown the effect removing the carotid body’s sympathetic innervation, via superior cervical ganglionectomy, will have on the carotid body’s chemoreflex response to hypoxia. We hypothesize that loss of sympathetic input will decrease the hypoxic ventilatory response (HVR) of the rat, and alter the O2‐sensing mechanism of the CB chemosensitive glomus cells. Sprague Dawley male rats postnatal day 21 were subjected to unilateral or bilateral superior cervical ganglionectomy (SCGX). Four days post surgery (post‐natal day 25), rats either underwent whole body plethysmography recordings or were sacrificed and their carotid bodies removed to either isolate glomus cells and perform whole cell perforated patch clamp, or perform immunohistochemistry on the CB. Our preliminary results show that unilateral (left or right) SCGX appears to decrease the HVR during a 1‐minute exposure to hypoxia (10% O2, 90% N2) compared to bilateral SCGX. Our results also show that following SCGX, isolated glomus cells responded to acute hypoxia (HX) (5–8% O2, 5% CO2, balanced N2) with significant reductions (P&lt;0.05 to P&lt;0.01) in outward K+ current amplitudes over the test potential range +10 to +60 mV. The mean maximal effect, a reduction in K+ amplitude of 64.9 ± 18.5% (mean ± SD, n = 3 cells), occurred at a Vtest of +30mV. In contrast, control carotid body glomus cells responded to acute HX with a significant reduction (P&lt;0.05) in outward K+ current amplitude at a Vtest of +60mV (n = 4 cells). In conclusion, our data suggests that sympathetic innervation to the CB is critical for the ventilatory response to hypoxia, and the O2‐sensing mechanism at the level of the CB glomus cells. Overall this study provides novel evidence for the importance of sympathetic innervation on peripheral chemosensors during exposure to hypoxia.Support or Funding InformationThis work was supported in part by a SPARC NIH award OT2OD023860

Carotid body‐specific shRNA knockdown of PKCɛ blunts TRPV1‐dependent asthmatic bronchoconstriction

RationaleAggravated airway bronchoconstriction and mucus production in response to allergen and/or bradykinin (i.e., airway hyperresponsiveness) is a primary defining symptom of asthma. Recently, we discovered a critical role for the carotid bodies: inhaled allergen causes an increase in arterial plasma concentrations of asthmakines which excite the carotid bodies via a transient receptor potential vanilloid 1 channel (TRPV1) dependent mechanism. Elevated carotid body excitation in turn increases efferent vagal activity, resulting in heightened bronchoconstriction. While our data demonstrate that the carotid bodies are strongly excited by multiple Th‐2 cytokines, in the ovalbumin primed and sensitized Brown Norway rat model of asthma, the primary asthmakine is likely lysophosphatidic acid (LPA). However, the cellular link between LPA receptors and TRPV1 has not been identified. Here we show that the effect of LPA on the carotid bodies is likely mediated by PKCɛ‐dependent phosphorylation of TRPV1.MethodsLentivirus vectors containing shRNA to knockdown PKCɛ or scrambled sequence were injected bilaterally into the carotid bodies of asthmatic rats (n=7, each). The responses of carotid bodies to LPA (5μM), and their subsequent responses to TRPV1 blockade (AMG9810, 10μM) were tested using the en bloc perfused carotid body preparation. The effects of carotid body‐specific PKCɛ knockdown on changes in lung mechanics in response to inhaled bradykinin were tested using the Flexivent system.ResultsCarotid bodies from asthmatic rats injected with PKCɛ knockdown sequence mounted smaller responses to LPA than those injected with scrambled sequence. No difference in LPA response was detected between groups following TRPV1 blockade, suggesting the primary target for PKCɛ is TRPV1. Asthmatic rats with carotid bodies injected with PKCɛ knockdown sequence had reduced lung resistance in response to bradykinin at 1, 10 and 20 minutes. Similar results were obtained using a PKCɛ inhibitory peptide.ConclusionWe show for the first time that carotid body‐mediated bronchoconstriction in asthmatic rats occurs through LPA receptor activation of PKCɛ and subsequent TRPV1‐mediated carotid sinus nerve excitation. Targeting carotid body LPA receptors, PKCɛ and/or TRPV1 may provide a new therapeutic target to reduce asthmatic airway hyperresponsiveness.Support or Funding InformationCIHR, Alberta Innovates, Alberta Lung Association and the Parker B. Francis Foundation.

Testosterone induces higher carotid body response to hypoxia in older adult male rats

Excessive carotid body responsiveness to chemical stimuli contributes to respiratory instability and apneas during sleep. In hypogonadal men, testosterone supplementation increases the risk of sleep‐disordered breathing; however, the site of action is unknown. The present study tested the hypothesis that exposure to elevated levels of testosterone potentiates the carotid body’s response to hypoxia. Because testosterone levels decline with age, we also determined whether these effects are age‐dependent. To do so, carotid bodies were isolated ex vivo from young (70 days old) and aging (185 days old) male Sprague‐Dawley rats. Isolated organs were maintained in oxygenated Tyrode solution (95% O2; 5% CO2) in the presence of testosterone (5 nM) or vehicle (&lt;0.001% DMSO) for 90–120 min. Preparations were then mounted in the recording chamber and a suction electrode was placed on the carotid sinus nerve to record activity (# of impulses/sec). Activity was recorded at rest and during exposure to hypoxia (0% O2, 5% CO2, 95% N2; 15 min). Basal carotid body activity did not differ between age groups; however, the responses to both stimuli were greater in young rats. Testosterone exposure augmented the responses to hypoxia in aging rats only. In vivo experiments performed on anesthetised rats show that testosterone injection (2 mg/kg) 30 min prior hypoxic exposure (12%O2; 20 min) potentiates the hypoxic ventilatory response in an age‐dependent fashion. We conclude that age‐dependent potentiation of the carotid body’s O2 chemosensitivity by testosterone may lead to ventilatory adjustments that exceed the organism’s need. In aging animals, this may predispose to respiratory instability and apnea during sleep.Support or Funding InformationSupported by a discovery grant from the Natural Sciences and Engineering Research Council of Canada (RK). Dr Janes was supported by a scholarship from the Réseau en Recherche Respiratoire du Québec.

Role of IP3 Receptors in Shaping the Carotid Chemoreceptor Response to Hypoxia But Not to Hypercapnia in the Rat Carotid Body: An Evidence Review.

This article addresses the disparity in the transduction pathways for hypoxic and hypercapnic stimuli in carotid body glomus cells. We investigated and reviewed the experimental evidence showing that the response to hypoxia, but not to hypercapnia, is mediated by 1,4,5-inositol triphosphate receptors (IP3R/s) regulating the intracellular calcium content [Ca2+]c in glomus cells. The rationale was based on the past observations that inhibition of oxidative phosphorylation leads to the explicit inhibition of the hypoxic chemoreflex. [Ca2+]c changes were measured using cellular Ca2+-sensitive fluorescent probes, and carotid sinus nerve (CSN) sensory discharge was recorded with bipolar electrodes in in vitro perfused-superfused rat carotid body preparations. The cell-permeant, 2-amino-ethoxy-diphenyl-borate (2-APB; 100μM) and curcumin (50μM) were used as the inhibitors of IP3R/s. These agents suppressed the [Ca2+]c, and CSN discharge increases in hypoxia but not in hypercapnia, leading to the conclusion that only the hypoxic effects were mediated via modulation of IP3R/s. The ATP-induced Ca2+ release from intracellular stores in a Ca2+-free medium was blocked with 2-APB, supporting this conclusion.

Molecular and cellular mechanisms of the organogenesis and development of the mammalian carotid body.

Despite significant advancements in understanding physiological properties of the carotid body, little attention has been paid to its organogenesis. This review addresses the molecular and cellular mechanisms underlying organogenesis of the carotid body in mammals. The carotid body consists of two types of cells, that is, glomus cells and sustentacular cells, that are derived from different origins. Glomus cells are derivatives of neural crest cells which form sympathetic ganglia. Sustentacular cells are derivatives of mesenchymal neural crest cells which colonize the third pharyngeal arch and form the wall of the third arch artery. Gene-targeting studies indicate that three elements are required for carotid body organogenesis: the carotid sinus nerve (CSN), third arch artery, and superior cervical sympathetic ganglion (SCG). The CSN sends sensory fibers and Schwann cells to the wall of the third arch artery. The third arch artery provides mesenchymal cells, which give rise to sustentacular cells. The nerve process from the SCG sends glomus cell progenitors into the carotid body primordium. The presence of stem cells in the adult carotid body was recently highlighted. The origin of stem cells, however, remains controversial. Based on embryonic development of the carotid body, this review proposes the origin of stem cells.

Carotid Body Baroreceptor Preservation and Control of Arterial Pressure in Eversion Carotid Endarterectomy.

The technique of the eversion carotid endarterectomy (ECEA), as an alternative to the conventional endarterectomy with primary or patch angioplasty, is an established technique for managing internal carotid artery stenoses and recently its application has been upgraded through the European Society for Vascular Surgery guidelines (Recommendation 55: Class 1, Level A). However, the typical eversion method has been associated with postoperative hypertension due to loss of the baroreceptor reflex; the standard oblique transection at the bulb performed in the eversion endarterectomy interrupts either the baroreceptor sensoring tissue, which is mostly located in the adventitia at the medial portion of the proximal internal carotid artery, or even the proper Hering nerve, a branch of the glossopharyngeal nerve. These actions deregulate the natural negative feedback of the carotid baroreceptor. Guided by the anatomical location of the baroreceptor sensor we have elaborated a slight modification of the classical ECEA to maintain as much as possible of the viable carotid baroreceptor sensoring surface. By extending the oblique incision distal to the carotid bifurcation in the medial part of the internal carotid artery stem, an eyebrow-like part of the proximal internal carotid artery is maintained and the axis from the sensoring tissue to the nerve of Hering is protected and following the endarterectomy, postoperative arterial blood pressure levels are lower than in the classical ECEA due to the maintenance of the efficiency of the baroreceptor reflex. During the period from September 2016 to November 2018, carotid endarterectomy was performed in 57 patients. Twenty-eight of them underwent the typical ECEA and 29 patients had the modified eyebrow eversion carotid endarterectomy (me-ECEA). The changes of blood pressure baseline during the postoperative course in ECEA and me-ECEA group were analyzed and compared. Postoperative hypertension was defined as an elevation of systolic blood pressure (SBP) greater than 140 mm Hg. Patients who underwent typical ECEA had significantly higher postoperative blood pressure values compared with those who underwent me-ECEA. Actually, the mean postoperative SBP was 172.67 ± 24.59 mm Hg in the typical ECEA group compared with 160.86 ± 12.83 mm Hg in the me-ECEA group ( p = 0.023). The mean diastolic blood pressure in the ECEA group was 65.42 ± 11.39 mm Hg compared with 58.06 ± 9.06 mm Hg in the me-ECEA group ( p = 0.009). Our proposed me-ECEA technique seems to be related to lower rates of postoperative hypertension compared with the typical ECEA, probably due to the sparing of the main mass of the baroreceptor apparatus; this improved modification (me-ECEA) of the typical eversion procedure could represent an alternative ECEA technique with its inherent advantages.

Feasibility of kilohertz frequency alternating current neuromodulation of carotid sinus nerve activity in the pig

Recent research supports that over-activation of the carotid body plays a key role in metabolic diseases like type 2 diabetes. Supressing carotid body signalling through carotid sinus nerve (CSN) modulation may offer a therapeutic approach for treating such diseases. Here we anatomically and histologically characterised the CSN in the farm pig as a recommended path to translational medicine. We developed an acute in vivo porcine model to assess the application of kilohertz frequency alternating current (KHFAC) to the CSN of evoked chemo-afferent CSN responses. Our results demonstrate the feasibility of this approach in an acute setting, as KHFAC modulation was able to successfully, yet variably, block evoked chemo-afferent responses. The observed variability in blocking response is believed to reflect the complex and diverse anatomy of the porcine CSN, which closely resembles human anatomy, as well as the need for optimisation of electrodes and parameters for a human-sized nerve. Overall, these results demonstrate the feasibility of neuromodulation of the CSN in an anesthetised large animal model, and represent the first steps in driving KHFAC modulation towards clinical translation. Chronic recovery disease models will be required to assess safety and efficacy of this potential therapeutic modality for application in diabetes treatment.

Assessing the Impact of the Sinus-Saving Modifications of Carotid Endarterectomy in the Autonomic Regulation and Central Hemodynamics

Introduction: There are many dangerous complications, such as myocardial infarction, hyperperfusion syndrome and its effects, hematomas in the postoperative period after carotid surgery. One of the main causes of these complications is the variation of central hemodynamic especially with the tendency to hypertension. Our goal is to develop an operative technique that could let us avoid the intersection of the carotid sinus nerves, which reduce the risk of complications thanks to a more manageable blood pressure, due to decreased sympathetic influence on the regulation of vascular tone. Methods: During our retrospective analysis 384 cases had been investigated. Prior to inclusion in the study, written informed consent was obtained from all participants. Patients were separated into two groups. Groups are even in age, sex, initial neurological and cardiac status and contralateral blood flow. The first group of patients (203 patients) had cut the carotid sinus nerves. Carotid sinus nerves of patients from the second group (181 patients) were kept safe. Evaluated the following parameters – systolic blood pressure, diastolic blood pressure, heart rate and pulse pressure. We are interested in the period – 1st and 4th days after surgery when the risk of complications and changes in hemodynamics is particularly high. Also on the 1st and 4th day after surgery, the state of the autonomic regulation was assessed by analyzing heart rate variability. We prospectively analyzed the autonomic regulation of 16 patients from the first group and 19 patients from the other one, observing a variety of heart rate at neurocardiological laboratory. Results: On the second postoperative day all the hemodynamic indicators (as SBP, DBP, pulse pressure, heart rate) had much lower levels in the second group of patients (p<0,05) with the recovery of normal numbers on the third day. This fact kept postoperative complications to a minimum. In the group of patients with saved carotid sinus nerve on the 1st day after surgery was more than a noticeable decrease sympathetic influence on the rhythm, with a tendency to restore autonomic regulation on the 4th day. These results explain the difference between central hemodynamic indicators in two groups. Conclusion: Obtained results show the lower activity of the sympathetic and the high activity of the parasympathetic system in the group with non-damaged carotid sinus nerves. The practical application of the sine-saving modification of eversion carotid endarterectomy allows reducing the risk of postoperative complications caused by arterial hypertension.

Sympathetic Effects of Internal Carotid Nerve Manipulation on Choroidal Vascularity and Related Measures.

To investigate specific effects of denervation and stimulation of the internal carotid nerve (ICN) on the choroid and retina. Female Sprague Dawley rats underwent unilateral ICN transection (n = 20) or acute ICN electrical stimulation (n = 7). Rats in the denervation group were euthanized 6 weeks after nerve transection, and eyes were analyzed for changes in choroidal vascularity (via histomorphometry) or angiogenic growth factors and inflammatory markers (via ELISA). Rats in the stimulation group received acute ICN electrical stimulation with a bipolar cuff electrode over a range of stimulus amplitudes, frequencies, and pulse widths. Choroidal blood flow and pupil diameter were monitored before, during, and after stimulation. Six weeks after unilateral ICN transection, sympathectomized choroids exhibited increased vascularity, defined as the percentage of choroidal surface area occupied by blood vessel lumina. Vascular endothelial growth factor (VEGF) and VEGF receptor-2 (VEGFR-2) protein levels in denervated choroids were 61% and 124% higher than in contralateral choroids, respectively. TNF-α levels in denervated retinas increased by 3.3-fold relative to levels in contralateral retinas. In animals undergoing acute ICN electrical stimulation, mydriasis and reduced choroidal blood flow were observed in the ipsilateral eye. The magnitude of the reduction in blood flow correlated positively with stimulus frequency. Modulation of ICN activity reveals a potential role of the ocular sympathetic system in regulating endpoints related to neovascular diseases of the eye.

Decoding Neural Metabolic Markers From the Carotid Sinus Nerve in a Type 2 Diabetes Model.

Recent studies showed that the carotid sinus nerve (CSN) and the sympathetic nervous system (SNS) are overactivated in type 2 diabetes and that restoring the correct CSN neural activity can re-establish the proper metabolism. However, a robust characterization of the relationship between CSN and SNS neural activities and metabolism in type 2 diabetes is still missing. Here, we investigated the relationship between neural activity of CSN and SNS in control rats and in rats with diet-induced type 2 diabetes and the animal condition during metabolic challenges. We found that the diabetic condition can be discriminated on the basis of CSN and SNS neural activities due to a high-frequency shift in both spectra. This shift is suppressed in the SNS in case of CSN denervation, confirming the role of CSN in driving sympathetic overactivation in type 2 diabetes. Interestingly, the Inter-Burst-Intervals (IBIs) calculated from CSN bursts strongly correlate with perturbations in glycaemia levels. This finding, held for both control and diabetic rats, indicates the possibility of detecting metabolic information from neural recordings even in pathological conditions. Our results suggest that CSN activity could serve as a marker to monitor glycaemic alterations and, therefore, it could be used for closed-loop control of CSN neuromodulation. This paves the way to the development of novel and effective bioelectronic therapies for type 2 diabetes.

Leptin Induces Hypertension Acting on Transient Receptor Potential Melastatin 7 Channel in the Carotid Body.

Obesity leads to resistant hypertension and mechanisms are poorly understood, but high plasma levels of leptin have been implicated. Leptin increases blood pressure acting both centrally in the dorsomedial hypothalamus and peripherally. Sites of the peripheral hypertensive effect of leptin have not been identified. We previously reported that leptin enhanced activity of the carotid sinus nerve, which transmits chemosensory input from the carotid bodies (CBs) to the medullary centers, and this effect was abolished by nonselective blockers of Trp (transient receptor potential) channels. We searched our mouse CB transcriptome database and found that the Trpm7 (transient receptor potential melastatin 7) channel was the most abundant Trp channel. To examine if leptin induces hypertension acting on the CB Trpm7. C57BL/6J (n=79), leptin receptor (LepRb) deficient db/db mice (n=22), and LepRb-EGFP (n=4) mice were used. CB Trpm7 and LepRb gene expression was determined and immunohistochemistry was performed; CB glomus cells were isolated and Trpm7-like current was recorded. Blood pressure was recorded continuously in (1) leptin-treated C57BL/6J mice with intact and denervated CB; (2) leptin-treated C57BL/6J mice, which also received a nonselective Trpm7 blocker FTY720 administered systemically or topically to the CB area; (3) leptin-treated C57BL/6J mice transfected with Trpm7 small hairpin RNA to the CB, and (4) Leprb deficient obese db/db mice before and after Leprb expression in CB. Leptin receptor and Trpm7 colocalized in the CB glomus cells. Leptin induced a nonselective cation current in these cells, which was inhibited by Trpm7 blockers. Leptin induced hypertension in C57BL/6J mice, which was abolished by CB denervation, Trpm 7 blockers, and Trpm7 small hairpin RNA applied to CBs. Leprb overexpression in CB of Leprb-deficient db/db mice demethylated the Trpm7 promoter, increased Trpm7 gene expression, and induced hypertension. We conclude that leptin induces hypertension acting on Trmp7 in CB, which opens horizons for new therapy.