Hypoxia In Carotid Body Research Articles

Neurochemical Plasticity of the Carotid Body.

A striking feature of the carotid body (CB) is its remarkable degree of plasticity in a variety of neurotransmitter/modulator systems in response to environmental stimuli, particularly following hypoxic exposure of animals and during ascent to high altitude. Current evidence suggests that acetylcholine and adenosine triphosphate are two major excitatory neurotransmitter candidates in the hypoxic CB, and they may also be involved as co-transmitters in hypoxic signaling. Conversely, dopamine, histamine and nitric oxide have recently been considered inhibitory transmitters/modulators of hypoxic chemosensitivity. It has also been revealed that interactions between excitatory and inhibitory messenger molecules occur during hypoxia. On the other hand, alterations in purinergic neurotransmitter mechanisms have been implicated in ventilatory acclimatization to hypoxia. Chronic hypoxia also induces profound changes in other neurochemical systems within the CB such as the catecholaminergic, peptidergic and nitrergic, which in turn may contribute to increased ventilatory and chemoreceptor responsiveness to hypoxia at high altitude. Taken together, current data suggest that complex interactions among transmitters markedly influence hypoxia-induced transmitter release from the CB. In addition, the expression of a wide variety of growth factors, proinflammatory cytokines and their receptors have been identified in CB parenchymal cells in response to hypoxia and their upregulated expression could mediate the local inflammation and functional alteration of the CB under hypoxic conditions.

Augmented Respiratory-Sympathetic Coupling and Hemodynamic Response to Acute Mild Hypoxia in Female Rodents With Chronic Kidney Disease.

Carotid body feedback and hypoxia may serve to enhance respiratory–sympathetic nerve coupling (respSNA) and act as a driver of increased blood pressure. Using the Lewis polycystic kidney (LPK) rat model of chronic kidney disease, we examined respSNA in adult female rodents with CKD and their response to acute hypoxia or hypercapnia compared to Lewis control animals. Under urethane anesthesia, phrenic nerve activity, splanchnic sympathetic nerve activity (sSNA), and renal sympathetic nerve activity (rSNA) were recorded under baseline conditions and during mild hypoxic or hypercapnic challenges. At baseline, tonic SNA and blood pressure were greater in female LPK rats versus Lewis rats (all P < 0.05) and respSNA was at least two-fold larger [area under the curve (AUC), sSNA: 7.8 ± 1.1 vs. 3.4 ± 0.7 μV s, rSNA: 11.5 ± 3 vs. 4.8 ± 0.7 μV s, LPK vs. Lewis, both P < 0.05]. Mild hypoxia produced a larger pressure response in LPK [Δ mean arterial pressure (MAP) 30 ± 6 vs. 12 ± 6 mmHg] and augmented respSNA (ΔAUC, sSNA: 8.9 ± 3.4 vs. 2 ± 0.7 μV s, rSNA: 6.1 ± 1.2 vs. 3.1 ± 0.7 μV s, LPK vs. Lewis, all P ≤ 0.05). In contrast, central chemoreceptor stimulation produced comparable changes in blood pressure and respSNA (ΔMAP 13 ± 3 vs. 9 ± 5 mmHg; respSNA ΔAUC, sSNA: 2.5 ± 1 vs. 1.3 ± 0.7 μV s, rSNA: 4.2 ± 0.9 vs. 3.5 ± 1.4 μV s, LPK vs. Lewis, all P > 0.05). These results demonstrate that female rats with CKD exhibit heightened respSNA coupling at baseline that is further augmented by mild hypoxia, and not by hypercapnia. This mechanism may be a contributing driver of hypertension in this animal model of CKD.

Guinea Pig as a Model to Study the Carotid Body Mediated Chronic Intermittent Hypoxia Effects.

Clinical and experimental evidence indicates a positive correlation between chronic intermittent hypoxia (CIH), increased carotid body (CB) chemosensitivity, enhanced sympatho-respiratory coupling and arterial hypertension and cardiovascular disease. Several groups have reported that both the afferent and efferent arms of the CB chemo-reflex are enhanced in CIH animal models through the oscillatory CB activation by recurrent hypoxia/reoxygenation episodes. Accordingly, CB ablation or denervation results in the reduction of these effects. To date, no studies have determined the effects of CIH treatment in chemo-reflex sensitization in guinea pig, a rodent with a hypofunctional CB and lacking ventilatory responses to hypoxia. We hypothesized that the lack of CB hypoxia response in guinea pig would suppress chemo-reflex sensitization and thereby would attenuate or eliminate respiratory, sympathetic and cardiovascular effects of CIH treatment. The main purpose of this study was to assess if guinea pig CB undergoes overactivation by CIH and to correlate CIH effects on CB chemoreceptors with cardiovascular and respiratory responses to hypoxia. We measured CB secretory activity, ventilatory parameters, systemic arterial pressure and sympathetic activity, basal and in response to acute hypoxia in two groups of animals: control and 30 days CIH exposed male guinea pigs. Our results indicated that CIH guinea pig CB lacks activity elicited by acute hypoxia measured as catecholamine (CA) secretory response or intracellular calcium transients. Plethysmography data showed that only severe hypoxia (7% O2) and hypercapnia (5% CO2) induced a significant increased ventilatory response in CIH animals, together with higher oxygen consumption. Therefore, CIH exposure blunted hyperventilation to hypoxia and hypercapnia normalized to oxygen consumption. Increase in plasma CA and superior cervical ganglion CA content was found, implying a CIH induced sympathetic hyperactivity. CIH promoted cardiovascular adjustments by increasing heart rate and mean arterial blood pressure without cardiac ventricle hypertrophy. In conclusion, CIH does not sensitize CB chemoreceptor response to hypoxia but promotes cardiovascular adjustments probably not mediated by the CB. Guinea pigs could represent an interesting model to elucidate the mechanisms that underlie the long-term effects of CIH exposure to provide evidence for the role of the CB mediating pathological effects in sleep apnea diseases.

Acute hypoxia alters reflex micturition behavior in urethane‐anesthetized carotid sinus intact and denervated adult female Sprague‐Dawley rat

Acute hypoxia is well known to increase respiratory drive by stimulating peripheral chemoreceptors located in the carotid bodies. Ongoing work in our laboratory has been examining the effects acute hypoxia on lower urinary tract (LUT) function in urethane‐anesthetized adult female rats. While these experiments have revealed that exposure to acute hypoxia is capable of modifying LUT function, the contribution of peripheral chemoreceptor‐mediated effects is unknown. Moreover, hypoxia‐induced modulation of LUT behavior is present both during hypoxia and transiently after the hypoxic stimulus is removed, such that hypoxia increases void frequency while the return to normoxia increases the threshold bladder pressure for voiding and prolongs the inter‐contraction interval (ICI). The present study was designed to evaluate whether the changes in LUT function noted are mediated by mechanisms that are dependent on carotid body hypoxia chemoreception. To this end, we conducted a series of experiments using continuous flow bladder cystometry with simultaneous recording of external urethral sphincter (EUS) EMG activity in urethane‐anesthetized spontaneously breathing adult female Sprague‐Dawley rats (n=10). Micturition events were elicited by saline infusion into the bladder via suprapubic catheter with the saline flow rate adjusted to induce a voiding frequency of approximately one void every 4–5 minutes. We also simultaneously recorded tongue, external oblique, and diaphragm muscle EMG activity via bipolar stainless steel EMG electrodes. Each rat was allowed to settle into a stable baseline voiding rhythm which was then recorded for approximately 1‐hr under normoxic conditions, after which an hypoxic challenge (12% O2, 15 min) was performed. In one group of rats, the carotid sinus nerves (CSN) were transected bilaterally prior to beginning the cytometry experiment while in a second group of rats the CSN were left intact. Success of the CSN denervation was confirmed by the absence of an increase in diaphragm EMG burst amplitude and frequency during the hypoxic challenge. Consistent with our prior observations, CSN intact rats responded to hypoxia with an increased voiding frequency, particularly at the onset of hypoxia; however, peak bladder pressure during contraction was variable with no consistent modulation by hypoxia. Upon return to normoxia, the initial voiding event was delayed, and subsequent events displayed increased ICI with elevated bladder pressure threshold. In contrast to CSN intact rats, CSN denervated rats exhibited a marked decrease in peak bladder contraction pressure during hypoxic exposure as well as a smaller increase in bladder pressure threshold after return to normoxia. No notable differences in hypoxia‐induced changes in ICI between CSN intact or transected rats was observed. These results suggest that hypoxia‐mediated carotid body afferent input is not required for producing a modulation of reflex micturition in response to hypoxia, but certain aspects of the hypoxic response may be impacted by carotid body dependent mechanisms.Support or Funding InformationNIH NS096514This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Oxygen sensing and stem cell activation in the hypoxic carotid body.

The carotid body (CB) is the major arterial chemoreceptor responsible for the detection of acute decreases in O2 tension (hypoxia) in arterial blood that trigger hyperventilation and sympathetic activation. The CB contains O2-sensitive glomus (chief) cells, which respond to hypoxia with the release of transmitters to activate sensory nerve fibers impinging upon the brain respiratory and autonomic centers. During exposure to sustained hypoxia (for weeks or months), the CB grows several-fold in size, a response associated with acclimatization to high altitude or to medical conditions presenting hypoxemia. Here, I briefly present recent advances on the mechanisms underlying glomus cell sensitivity to hypoxia, in particular the role of mitochondrial complex I in acute oxygen sensing. I also summarize the properties of adult CB stem cells and of glomus cell-stem cell synapses, which contribute to CB hypertrophy in chronic hypoxia. A note on the relationship between hypoxic CB growth and tumorigenesis is included. Finally, the medical implications of CB pathophysiology are discussed.

Neonatal Maternal Separation Augments Carotid Body Response to Hypoxia in Adult Males but Not Female Rats.

Perinatal exposure to adverse experiences disrupts brain development, including the brainstem network that regulates breathing. At adulthood, rats previously subjected to stress (in the form of neonatal maternal separation; NMS) display features reported in patients suffering from sleep disordered breathing, including an increased hypoxic ventilatory response and hypertension. This effect is also sex-specific (males only). Based on these observations, we hypothesized that NMS augments the carotid body's O2-chemosensitivity. Using an isolated and perfused ex vivo carotid body preparation from adult rats we compared carotid sinus nerve (CSN) responses to hypoxia and hypercapnia in carotid bodies harvested from adult rats that either experienced control conditions (no experimental manipulation) or were subjected to NMS (3 h/day from postnatal days 3 to 12). In males, the CSN response to hypoxia measured in preparations from NMS males was 1.5 fold higher than controls. In control rats, the female's response was similar to that of males; however, the increase in CSN activity measured in NMS females was 3.0 times lower than controls. The CSN response to hypercapnia was not influenced by stress or sex. We conclude that NMS is sufficient to have persistent and sex-specific effects on the carotid body's response to hypoxia. Because NMS also has sex-specific effects on the neuroendocrine response to stress, we propose that carotid body function is influenced by stress hormones. This, in turn, leads to a predisposition toward cardio-respiratory disorders.

Measurement of ROS Levels and Membrane Potential Dynamics in the Intact Carotid Body Ex Vivo.

Reactive oxygen species (ROS) generated by the NADPH oxidase have been proposed to play an important role in the carotid body (CB) oxygen sensing process (Cross et al. 1990). Up to now it remains unclear whether hypoxia causes an increase or decrease of CB ROS levels. We transfected CBs with the ROS sensitive HSP-FRET construct and subsequently measured the intracellular redox state by means of Förster resonance energy transfer (FRET) microscopy. In a previous study we found both increasing and decreasing ROS levels under hypoxic conditions. The transition from decreasing to increasing ROS levels coincided with the change of the caging system from ambient environment caging (AEC) to individually ventilated caging (IVC) (Bernardini A, Brockmeier U, Metzen E, Berchner-Pfannschmidt U, Harde E, Acker-Palmer A, Papkovsky D, Acker H, Fandrey J, Type I cell ROS kinetics under hypoxia in the intact mouse carotid body ex vivo: a FRET based study. Am J Physiol Cell Physiol. doi: 10.1152/ajpcell.00370.2013 , 2014). In this work we analyze hypoxia induced ROS reaction of animals from an IVC system that had been exposed to AEC conditions for 5 days. The results further support the hypothesis of an important impact of the caging system on CB ROS reaction.

Inhibition of Protein Kinases AKT and ERK1/2 Reduce the Carotid Body Chemoreceptor Response to Hypoxia in Adult Rats.

The carotid body is the main mammalian oxygen-sensing organ regulating ventilation. Despite the carotid body is subjected of extensive anatomical and functional studies, little is yet known about the molecular pathways signaling the neurotransmission and neuromodulation of the chemoreflex activity. As kinases are molecules widely involved in motioning a broad number of neural processes, here we hypothesized that pathways of protein kinase B (AKT) and extracellular signal-regulated kinases ½ (ERK1/2) are implicated in the carotid body response to hypoxia. This hypothesis was tested using the in-vitro carotid body/carotid sinus nerve preparation ("en bloc") from Sprague Dawley adult rats. Preparations were incubated for 60 min in tyrode perfusion solution (control) or containing 1 μM of LY294002 (AKT inhibitor), or 1 μM of UO-126 (ERK1/2 inhibitor). The carotid sinus nerve chemoreceptor discharge rate was recorded under baseline (perfusion solution bubbled with 5 % CO(2) balanced in O(2)) and hypoxic (perfusion solution bubbled with 5 % CO(2) balanced in N(2)) conditions. Compared to control, both inhibitors significantly decreased the normoxic and hypoxic carotid body chemoreceptor activity. LY294002- reduced carotid sinus nerve discharge rate in hypoxia by about 20 %, while UO-126 reduces the hypoxic response by 45 %. We concluded that both AKT and ERK1/2 pathways are crucial for the carotid body intracellular signaling process in response to hypoxia.

Type I cell ROS kinetics under hypoxia in the intact mouse carotid body ex vivo: a FRET-based study.

Reactive oxygen species (ROS) mainly originating from NADPH oxidases have been shown to be involved in the carotid body (CB) oxygen-sensing cascade. For measuring ROS kinetics, type I cells of the mouse CB in an ex vivo preparation were transfected with the ROS sensor construct FRET-HSP33. After 2 days of tissue culture, type I cells expressed FRET-HSP33 as shown by immunohistochemistry. In one population of CBs, 5 min of hypoxia induced a significant and reversible decrease of type I cell ROS levels (n = 9 CBs; P < 0.015), which could be inhibited by 4-(2-aminoethyl)benzensulfonylfluorid (AEBSF), a highly specific inhibitor of the NADPH oxidase subunits p47(phox) and p67(phox). In another population of CBs, however, 5 min of hypoxia induced a significant and reversible increase of ROS levels in type I cells (n = 8 CBs; P < 0.05), which was slightly enhanced by administration of 3 mM AEBSF. These different ROS kinetics seemed to coincide with different mice breeding conditions. Type I cells of both populations showed a typical hypoxia-induced membrane potential (MP) depolarization, which could be inhibited by 3 mM AEBSF. ROS and MP closely followed the hypoxic decrease in CB tissue oxygen as measured with an O2-sensitive dye. We conclude that attenuated p47(phox) subunit activity of the NADPH oxidase under hypoxia is the physiological trigger for type I cell MP depolarization probably due to ROS decrease, whereas the observed ROS increase has no influence on type I cell MP kinetics under hypoxia.

Enhanced adenosine A2b receptor signaling facilitates stimulus-induced catecholamine secretion in chronically hypoxic carotid body type I cells

Chronic hypoxia (CHox) augments chemoafferent activity in sensory fibers innervating carotid body (CB) chemoreceptor type I cells; however, the underlying mechanisms are poorly understood. We tested the hypothesis that enhanced paracrine signaling via adenosine (Ado) A2b receptors is involved. Dissociated rat CB cultures were exposed for 24 h to normoxia (Nox, 21% O2) or CHox (2% O2) or treated with the hypoxia mimetic deferoxamine mesylate (DFX), and catecholamine secretion from type I cells was monitored by amperometry. Catecholamine secretion was more robust in CHox and DFX type I cells than Nox controls after acute exposure to acid hypercapnia (10% CO2, pH 7.1) and high K(+) (75 mM). Exogenous Ado increased catecholamine secretion in a dose-dependent manner, and the EC50 was shifted to the right from ∼21 μM Ado in Nox cells to ∼78 μM in CHox cells. Ado-evoked secretion in Nox and CHox cells was markedly inhibited by MRS-1754, an A2b receptor blocker, but was unaffected by SCH-58261, an A2a receptor blocker. Similarly, MRS-1754, but not SCH-58261, partially inhibited high-K(+)-evoked catecholamine secretion, suggesting a contribution from paracrine activation of A2b receptors by endogenous Ado. CB chemostimuli, acid hypercapnia, and hypoxia elicited a MRS-1754-sensitive rise in intracellular Ca(2+) that was more robust in CHox and DFX than Nox cells. Taken together, these data suggest that paracrine Ado A2b receptor signaling contributes to stimulus-evoked catecholamine secretion in Nox and CHox CB chemoreceptors; however, the effects of Ado are more robust after CHox.

NAD(P)H autofluorescence induction by Compound C in rat carotid chemoreceptor cells

Compound C (CC) has been known as an inhibitor of AMP‐activated kinase (AMPK). Recently other roles of CC have been reported. CC inhibits hypoxia dependent HIF‐1α activation independent of AMPK but dependent on mitochondrial electron transport chain (ETC) (Emerling 07) and CC functions as an inhibitor of complex I of ETC (Chua 11). In addition, we have found that CC (4μM) produces ~10 fold increase in NAD(P)H autofluorescence (AutoFlu) as compared to hypoxia or cyanide in rat carotid body (CB) glomus cells. Autoflu was detected with 360/50 excitation(x) and 460/50 emission(m) filters. The effect of CC on the hypoxic [Ca2+]i response was observed by using 340&amp;380x with 540/50m filter for fura‐2. Rat CB glomus cells were identified with rhodamine‐tagged peanut agglutinin.Results1) AutoFlu produced by CC is dose dependent, reaching a plateau at 0.04μM in both CB type cells. 2) The magnitude of AutoFlu induced by CC was greater in glomus cells compared to non‐glomus cells. 3) AutoFlu induced by 0.4μM CC was reduced ~50% by hypoxia but only &lt; 10% by 20mM KCl. 4) CC did not increase calcium despite the increase in AutoFlu. 5) 0.4μM CC reduced the hypoxia‐induced [Ca2+]i increase by ~50%, but did not block 20mM KCl‐induced [Ca2+]i increase. These preliminary results suggest that CC induced significant NAD(P)H AutoFlu, which may significantly affect the measured [Ca2+]i response to hypoxia in glomus cells. (Funded by NIH HL54621)

Activation of 5HT1A receptors in the nucleus tractus solitarius contributes to blood pressure compensation with blood loss and peripheral chemoreflex responses in the presence of acidosis

Caudal serotonin neurons and the 5‐HT1A receptor ( 1AR) agonist, 8‐OH‐DPAT, promote sympathetic‐mediated increases in cardiac output after blood loss. We hypothesize that the metabolic acidosis of hemorrhage activates serotonin neurons and 1AR in the nucleus tractus solitarius (NTS) to facilitate chemoreflex responses to carotid body hypoxia. Here, we tested ventilatory responses to hypoxic hypercapnia (10% O2, 5% CO2) and blood pressure responses to hemorrhage and subsequent 8‐OH‐DPAT administration in rats subjected to bilateral knockdown of the 1AR in the NTS. An adeno‐associated viral vector harboring a short hairpin RNA targeting the rat 1AR (1ARshRNA) or a scrambled shRNA (Scram) was injected into the caudal NTS of male Sprague Dawley rats. After 4 weeks, rats injected with the 1AshRNA (n=12) had lower cumulative minute ventilation during hypoxic hypercapnia vs. Scram‐injected rats (n=13, P&lt;0.01). Rats injected with 1AshRNA (n=8) had a more rapid fall in blood pressure with blood loss and an attenuated blood pressure response to 8‐OH‐DPAT injection (30 nmol/kg, iv, P&lt;0.01) vs. Scram‐injected rats (n=9). 1AshRNA injection decreased 1AR protein in the caudal NTS (−35%, P&lt;0.05). Thus, the beneficial hemodynamic effect of 8‐OH‐DPAT and endogenous serotonin in hemorrhage is related to activation of 1AR in the NTS and possibly to facilitation of chemoreflex responses to hypoxia. Funded by AHA 10PRE4000023.

Changes in oxygen sensitivity of TASK in carotid body glomus cells during early postnatal development

A post-natal increase in carotid body (CB) hypoxia responsiveness occurs at the level of carotid sinus nerve activity, intracellular calcium, cell membrane depolarization and hypoxic inhibition of O(2)-sensitive background K(+) conductance. TASK-1, TASK-1/3 and TASK-3 are functionally expressed in CB glomus cells, with TASK-1/3 providing the major part of the O(2)-sensitive TASK-like background K(+) conductance. Here we report the effects of graded hypoxia on TASK-like channel activity in CB glomus cells from rats aged 0 to 1, 6 to 7 and 16 to 18 days; the time frame of postnatal CB functional maturation. TASK was active in nearly all cell-attached patches and TASK activity during normoxia did not differ across ages. Hypoxia produced a progressive decrease in channel opening frequency with graded decreases in O(2) level and also produced glomus cell depolarization, as assessed by the shift in reversal potential of TASK single channel current. Hypoxic inhibition of TASK activity was least at P0-P1 and increased with age mainly between 6-7 and 16-18 days. The O(2)-sensitive TASK activity was significantly greater in glomus cells from P16 to P18 when compared to cells from P0 to P1 day old rats. These results support the hypothesis that postnatal carotid body functional maturation is due, at least in part, to changes in the sensitivity of TASK to the hypoxic signals generated in glomus cells.

Role of MaxiK-type calcium dependent K+ channels in rat carotid body hypoxia transduction during postnatal development

Carotid body chemoreceptors transduce a decrease in arterial oxygen tension into increased sinus nerve action potential (AP) activity which undergoes a maturational increase in the post-natal period. MaxiK-channels channels are proposed to play a major role in organ function based on their maturation-dependent expression in glomus cells and inhibition by acute hypoxia. To better resolve the role of this channel, single-unit AP activity of rat chemoreceptor neurons was recorded, in vitro, during a progressive decrease in oxygen from normoxia (∼150 Torr) to moderate hypoxia (∼60 Torr). Blockade of MaxiK channels with charybdotoxin (100 nM) in both older (P16-P18) and younger (P2-P3) animals resulted in no significant change in AP activity, but increased nerve conduction speed in the older animals. In dissociated glomus cells, charybdotoxin slightly enhanced the intracellular calcium response to acute hypoxia at both ages. We conclude that MaxiK channels play little or no role in mediating the response to acute, moderate hypoxia, either in the newborn or older animal.

Differences in tyrosine hydroxylase expression after short-term hypoxia, hypercapnia or hypercapnic hypoxia in rat carotid body

In the carotid body (CB), it has been reported that the expressions of tyrosine hydroxylase (TH) mRNA and TH protein are enhanced by exposure to hypoxia. However, it is not known whether CO(2) affects the expression of TH in the CB. We examined the expression of TH mRNA and the immunoreactivity for TH in the CB of rats exposed to hypoxia (10% O(2)), hypercapnia (10% CO(2)) and hypercapnic hypoxia (10% O(2) and 10% CO(2)) for 2-24 h. The expression of TH mRNA in the CB was markedly enhanced in rats exposed to hypoxia for 4 h (6.6-fold), 6 h (6.0-fold) and 8 h (7.8-fold), and in rats exposed to hypercapnic hypoxia for 12 h (4.8-fold). The most intense TH immunoreactivity was observed in the CB from rats exposed to hypoxia for 12 and 24 h and to hypercapnic hypoxia for 24 h. The expressions of TH mRNA and the immunoreactivity for TH were not altered in the CB of rats exposed to hypercapnia. It is suggested that CO(2) does not affect TH expression in the CB, and that it inhibits hypoxia-enhanced TH expression.

A negative interaction between brainstem and peripheral respiratory chemoreceptors modulates peripheral chemoreflex magnitude

Interaction between central (brainstem) and peripheral (carotid body) respiratory chemosensitivity is vital to protect blood gases against potentially deleterious fluctuations, especially during sleep. Previously, using an in situ arterially perfused, vagotomized, decerebrate preparation in which brainstem and peripheral chemoreceptors are perfused separately (i.e. dual perfused preparation; DPP), we observed that the phrenic response to specific carotid body hypoxia was larger when the brainstem was held at 25 Torr P(CO(2)) compared to 50 Torr P(CO(2)). This suggests a negative (i.e. hypo-additive) interaction between chemoreceptors. The current study was designed to (a) determine whether this observation could be generalized to all carotid body stimuli, and (b) exclude the possibility that the hypo-additive response was the simple consequence of ventilatory saturation at high brainstem P(CO(2)). Specifically, we tested how steady-state brainstem P(CO(2)) modulates peripheral chemoreflex magnitude in response to carotid body P(CO(2)) and P(O(2)) perturbations, both above and below eupnoeic levels. We found that the peripheral chemoreflex was more responsive the lower the brainstem P(CO(2)) regardless of whether the peripheral chemoreceptors received stimuli which increased or decreased activation. These findings demonstrate a negative interaction between brainstem and peripheral chemosensitivity in the rat in the absence of ventilatory saturation. We suggest that a negative interaction in humans may contribute to increased controller gain associated with sleep-related breathing disorders and propose that the assumption of simple addition between chemoreceptor inputs used in current models of the respiratory control system be reconsidered.

Glia-like Stem Cells Sustain Physiologic Neurogenesis in the Adult Mammalian Carotid Body

Neurogenesis is known to occur in the specific niches of the adult mammalian brain, but whether germinal centers exist in the neural-crest-derived peripheral nervous system is unknown. We have discovered stem cells in the adult carotid body (CB), an oxygen-sensing organ of the sympathoadrenal lineage that grows in chronic hypoxemia. Production of new neuron-like CB glomus cells depends on a population of stem cells, which form multipotent and self-renewing colonies in vitro. Cell fate mapping experiments indicate that, unexpectedly, CB stem cells are the glia-like sustentacular cells and can be identified using glial markers. Remarkably, stem cell-derived glomus cells have the same complex chemosensory properties as mature in situ glomus cells. They are highly dopaminergic and produce glial cell line-derived neurotrophic factor. Thus, the mammalian CB is a neurogenic center with a recognizable physiological function in adult life. CB stem cells could be potentially useful for antiparkinsonian cell therapy.

Brainstem PCO2 modulates phrenic responses to specific carotid body hypoxia in an in situ dual perfused rat preparation

Inputs from central (brainstem) and peripheral (carotid body) respiratory chemoreceptors are coordinated to protect blood gases against potentially deleterious fluctuations. However, the mathematics of the steady-state interaction between chemoreceptors has been difficult to ascertain. Further, how this interaction affects time-dependent phenomena (in which chemoresponses depend upon previous experience) is largely unknown. To determine how central P(CO2) modulates the response to peripheral chemostimulation in the rat, we utilized an in situ arterially perfused, vagotomized, decerebrate preparation, in which central and peripheral chemoreceptors were perfused separately (i.e. dual perfused preparation (DPP)). We carried out two sets of experiments: in Experiment 1, we alternated steady-state brainstem P(CO2) between 25 and 50 Torr in each preparation, and applied specific carotid body hypoxia (60 Torr P(O2) and 40 Torr P(CO2)) under both conditions; in Experiment 2, we applied four 5 min bouts (separated by 5 min) of specific carotid body hypoxia (60 Torr P(O2) and 40 Torr P(CO2)) while holding the brainstem at either 30 Torr or 50 Torr P(CO2). We demonstrate that the level of brainstem P(CO2) modulates (a) the magnitude of the phrenic responses to a single step of specific carotid body hypoxia and (b) the magnitude of time-dependent phenomena. We report that the interaction between chemoreceptors is negative (i.e. hypo-additive), whereby a lower brainstem P(CO2) augments phrenic responses resulting from specific carotid body hypoxia. A negative interaction may underlie the pathophysiology of central sleep apnoea in populations that are chronically hypocapnic.

Role of NADPH oxidase‐derived superoxide anion on angiotensin II‐enhanced sensitivity of potassium channels to hypoxia in carotid body of congestive heart failure rabbits

Our previous study has shown that endogenous angiotensin II (Ang II) via AT1 receptor activation is involved in the enhanced sensitivity of K+ currents (IK) to hypoxia in carotid body (CB) glomus cells in congestive heart failure (CHF)(J Physiol, 575:–227; 2006). A significant body of evidence indicates that NADPH oxidase-derived superoxide anion mediates the effects of Ang II. We investigated whether NADPH oxidase-derived superoxide anion is a mediator of this Ang II effect in CB glomus cells, using the conventional whole-cell patch clamp technique. Ang II augmented the effect of hypoxia on IK (46.2 ± 4.1% inhibition vs 22.5 ± 3.9% inhibition in control) and L158,809 (1 μM, AT1 receptor antagonist), phenylarsine oxide (2 μM, PAO, NADPH oxidase inhibitor) and tempol (1 mM, superoxide scavenger) significantly blunted the effect of Ang II on sensitivity of IK to hypoxia in CB glomus cells from sham rabbits (21.7 ± 3.4%, 21.3 ± 2.7%, 20.6 ± 2.9% inhibition vs 46.2 ± 4.1% inhibition for Ang II alone, p<0.05). In CHF rabbits, the effect of hypoxia on IK in CB glomus cells was greater (53.5 ± 4.4% inhibition) than sham, and L158, 809, PAO and tempol markedly decreased the hypoxic inhibition of IK (27.7 ± 3.8%, 27.2 ± 3.2%, 26.9 ± 3.5% inhibition, p<0.05). These results indicate that the NADPH oxidase-superoxide anion contributes to the Ang II-enhanced sensitivity of K+ channels to hypoxia in CB glomus cells from CHF rabbits.

Acute Oxygen Sensing in Heme Oxygenase-2 Null Mice

Hemeoxygenase-2 (HO-2) is an antioxidant enzyme that can modulate recombinant maxi-K+ channels and has been proposed to be the acute O2 sensor in the carotid body (CB). We have tested the physiological contribution of this enzyme to O2 sensing using HO-2 null mice. HO-2 deficiency leads to a CB phenotype characterized by organ growth and alteration in the expression of stress-dependent genes, including the maxi-K+ channel α-subunit. However, sensitivity to hypoxia of CB is remarkably similar in HO-2 null animals and their control littermates. Moreover, the response to hypoxia in mouse and rat CB cells was maintained after blockade of maxi-K+ channels with iberiotoxin. Hypoxia responsiveness of the adrenal medulla (AM) (another acutely responding O2-sensitive organ) was also unaltered by HO-2 deficiency. Our data suggest that redox disregulation resulting from HO-2 deficiency affects maxi-K+ channel gene expression but it does not alter the intrinsic O2 sensitivity of CB or AM cells. Therefore, HO-2 is not a universally used acute O2 sensor.