Central Chemoreceptors Research Articles

Respiratory Responses to Intravenous Infusion of Sodium Lactate in Male and Female Wistar Rats

In patients with panic disorder or premenstrual dysphoria, anxiety attacks can be triggered by intravenous administration of sodium lactate. Since respiratory symptoms, such as hyperventilation and shortness of breath, are characteristic features of spontaneous as well as lactate-induced panic, an involvement of central or peripheral chemoreceptors in this reaction has been suggested. In the present study, we examined to what extent intravenous infusion of sodium lactate influences respiratory parameters in freely moving male and female Wistar rats. Prompted by clinical reports suggesting that the susceptibility to spontaneous and lactate-induced anxiety may be influenced by the menstrual cycle, we also investigated if the effect of lactate on respiration in female rats is estrus cycle-dependent. Male and ovariectomized female rats exposed to sodium lactate displayed a larger increase in respiratory rate than rats given an infusion of saline. In intact female rats, the response to lactate infusion was significantly more pronounced in the diestrus phase than in the proestrus/estrus phase of the cycle. It is concluded that sodium lactate is a respiratory stimulant in rat, and that this effect is influenced by female sex steroids.

Extrabranchial chemoreceptors involved in respiratory reflexes in the neotropical fish Colossoma macropomum (the tambaqui).

In a previous study, complete denervation of the gills in the tambaqui Colossoma macropomum did not eliminate the increase in breathing amplitude seen during exposure of this species to hypoxia. The present study was designed to examine other sites of putative O(2)-sensitive receptors that could be involved in this reflex action. Superfusion of the exposed brain of decerebrate, spinalectomized fish did not reveal the presence of central chemoreceptors responsive to hyperoxic, hypoxic, hypercarbic, acidic or alkaline solutions. Subsequent central transection of cranial nerve IX and X, removing not only all innervation of the gills but also sensory input from the lateral-line, cardiac and visceral branches of the vagus nerve, did not eliminate the increase in breathing amplitude that remained following peripheral gill denervation alone. Administration of exogenous catecholamines (10 and 100 nmol kg(-1) adrenaline) to fish with intact brains and minimal surgical preparation reduced both respiratory frequency and amplitude, suggesting that humoral release of adrenaline also could not be responsible for the increase in breathing amplitude that remained following gill denervation. Denervation of the mandibular branches of cranial nerve V and the opercular and palatine branches of cranial nerve VII in gill-denervated fish (either peripheral gill denervation or central section of cranial nerves IX and X), however, did eliminate the response. Thus, our data suggest that hypoxic and hyperoxic ventilatory responses as well as ventilatory responses to internal and external injections of NaCN in the tambaqui arise from O(2)-sensitive receptors in the orobranchial cavity innervated by cranial nerves V and VII and O(2)-sensitive receptors on the gills innervated by cranial nerves IX and X. Our results also revealed the presence of receptors in the gills that account for all of the increase in ventilation amplitude and part of the increase in ventilation frequency during hyperoxic hypercarbia, a group or groups of receptors, which may be external to the orobranchial cavity (but not in the central nervous system), that contribute to the increase in ventilation frequency seen in response to hyperoxic hypercarbia and the possible presence of CO(2)-sensitive receptors that inhibit ventilation frequency, possibly in the olfactory epithelium.

The Ventilatory Response to Cholecystokinin Tetrapeptide in Healthy Volunteers

The cholecystokinin (CCK) system, which has been shown to interact with both the panicogenic and respiratory systems, provides an interesting mechanism to further evaluate the central chemoreceptor and its effect on panic attack sensitivity. Intravenous CCK, a naturally occurring neuropeptide in the brain, has been found to induce the emotional and somatic symptoms of panic in both Panic Disorder (PD) and Normal Control (NC) subjects in a dose-dependent and reproducible fashion. To induce these effects, lower doses of intravenous CCK are required in the PD patients, relative to the NC subjects potentially suggesting that endogenous alterations in the CCK system may be contributing to the development of PD.Intravenous administration of CCK-4 in association with panic also results in subjective dyspnea, that is, diminution in vital capacity without an effect on the respiratory resistance. CCK-4 also causes a significant increase in tidal volume and minute ventilation but has no effect on breathing frequency. These observations suggest that a CCK-B receptor agonist may be acting as a respiratory stimulant, exerting its effect on anxiety through a direct effect on respiration.This study represents an examination of the specific effects of CCK-4 on the central chemoreceptor response.The study used a modified rebreathing technique, which accurately measures the ventilatory response to carbon dioxide in terms of both threshold and sensitivity. This technique requires the subject to rebreathe from a bag containing a hyperoxic and hypercapnic gas mixture resulting in rapid equilibration between alveolar gas and arterial blood. Use of a hyperoxic gas allows for the preferential examination of the central chemoreflexes (sensitive to CO2) with little if any effect of the peripheral (oxygen sensitive) chemoreflexes.After significant training, 15 healthy control subjects were assigned via a double blind procedure to receive an intravenous injection of placebo or CCK-4, using a between-subjects design. A between-subjects comparison was undertaken for the injection run (run #3) between subjects receiving the CCK-4 injection and those receiving the placebo injection. As well, a within-subject comparison was undertaken to compare the results of the run following the injection (run #3) vs. the previous run when no injection took place (run #2).No significant differences were noted between subjects who received CCK-4 as compared with placebo for: basal or sub-threshold ventilation, threshold CO2 resulting in a change in ventilation, or sensitivity of the central chemoreflex, regardless of whether a panic attack did or did not take place. In addition, within the group receiving the CCK-4 challenge, no significant differences were noted during run #3 (received the CCK-4 injection) and a prior run where no injection took place (run #2).We conclude that CCK-4 does not act to induce panic by altering the central (CO2 sensitive) chemoreceptor.

CO(2)-induced c-Fos expression in brainstem preprotachykinin mRNA containing neurons.

Tachykinin peptides are found in brainstem regions involved in central chemoreception and they may play a modulatory role in ventilatory response to hypercapnia. We determined whether tachykinin peptide containing neurons are activated by CO(2) by combining in situ hybridization and immunohistochemistry (IHH). Experiments were performed in 21-day-old rats exposed to 12% CO(2) for 1 h. c-Fos expression was identified by IHH on free floating sections (40 microm) that were mounted and then hybridized with anti-sense 35S labeled ribonucleotide probe of the rat preprotachykinin A (PPT-A) gene. Sections were analyzed for expression of the PPT-A gene, c-Fos protein and colocalization of PPT-A gene with c-Fos protein. Within the chemosensory region of the nucleus tractus solitarius (nTS), 19% of c-Fos positive cells expressed PPT-A mRNA after hypercapnic loading. In medullary raphe nuclei, 64% of c-Fos positive cells expressed the PPT-A gene after exposure to CO(2), while 21% of c-Fos labeled neurons in parapyramidal nuclei also expressed PPT-A mRNA. These results indicate that a subpopulation of CO(2) activated neurons within the nTS and in the parapyramidal and midline regions of the ventral aspect of the medulla oblongata express the PPT-A gene, suggesting that these are substance P- or neurokinin A-containing neurons. Furthermore, these peptides may play a role in modulation of respiratory and cardiovascular responses to changes in CO(2)/H(+) content of the extracellular fluid.

Ventilatory sensitivity to changes in inspired and arterial carbon dioxide partial pressures in the chicken

To determine if intrapulmonary chemoreceptors could be the sole peripheral chemoreceptors responsible for ventilatory responses to inhaled CO2, we studied the relationships of minute ventilation, tidal volume, and respiratory frequency to inspired and arterial partial pressure of CO2 (P(I)CO2 and PaCO2) in decerebrate upright chickens when the birds inspired gases containing four low partial pressures of CO2 (0.02, 5.0, 8.2, and 11.4 torr). Because of variability in the measured variables from time to time in the same birds, as well as between birds, and because of the limited precision in measuring PaCO2, a 4 × 4 Latin square design and four statistical methods of data analyses (modified reduced major axis estimator, maximum likelihood estimator, average slope method, and summary slope method) were used. Tidal volume, minute ventilation, and PaCO2 increased, but respiratory frequency remained unchanged, as gases containing increased partial pressures of CO2 were inhaled. The results indicate that intrapulmonary chemoreceptors are not the sole receptors stimulated by inhaling gas containing even low partial pressures of CO2 and that stimulation of arterial and central chemoreceptors also occurs. Stimulation of these latter chemoreceptors may account for the increase in ventilation. Further, the results demonstrate the importance of maintaining a low level of environmental CO2 in poultry houses to minimize its influence on ventilation.

Quantification of the response of rat medullary raphe neurones to independent changes in pH(o) and P(CO2).

The medullary raphe nuclei contain putative central respiratory chemoreceptor neurones that are highly sensitive to acidosis. To define the primary stimulus for chemosensitivity in these neurones, the response to hypercapnic acidosis was quantified and compared with the response to independent changes in P(CO2) and extracellular pH (pH(o)). Neurones from the ventromedial medulla of neonatal rats (P0-P2) were dissociated and maintained in tissue culture for long enough to develop a mature response (up to 70 days). Perforated patch clamp recordings were used to record membrane potential and firing rate while changes were made in pH(o), P(CO2) and/or [NaHCO(3)](o) from baseline values of 7.4, 5 % and 26 mM, respectively. Hypercapnic acidosis (P(CO2) 9 %; pH(o) 7.17) induced an increase in firing rate to 285 % of control in one subset of neurones ('stimulated neurones') and induced a decrease in firing rate to 21 % of control in a different subset of neurones ('inhibited neurones'). Isocapnic acidosis (pH(o) 7.16; [NaHCO(3)](o) 15 mM) induced an increase in firing rate of stimulated neurones to 309 % of control, and a decrease in firing rate of inhibited neurones to 38 % of control. In a different group of neurones, isohydric hypercapnia (9 % P(CO2); [NaHCO(3)](o) 40 mM) induced an increase in firing rate of stimulated neurones by the same amount (to 384 % of control) as in response to hypercapnic acidosis (to 327 % of control). Inhibited neurones also responded to isohydric hypercapnia in the same way as they did to hypercapnic acidosis. In Hepes-buffered solution, both types of neurone responded to changes in pH(o) in the same way as they responded to changes in pH(o) in bicarbonate-buffered Ringer solution. It has previously been shown that all acidosis-stimulated neurones in the medullary raphe are immunoreactive for tryptophan hydroxylase (TpOH-ir). Here it was found that TpOH-ir neurones in the medullary raphe were immunoreactive for carbonic anhydrase type II and type IV (CA II and CA IV). However, CA immunoreactivity was also common in neurones of the hypoglossal motor nucleus, inferior olive, hippocampus and cerebellum, indicating that its presence is not uniquely associated with chemosensitive neurones. In addition, under the conditions used here, acetazolamide (100 microM) did not have a significant effect on the response to hypercapnic acidosis. We conclude that chemosensitivity of raphe neurones can occur independently of changes in pH(o), P(CO2) or bicarbonate. The results suggest that a change in intracellular pH (pH(i)) may be the primary stimulus for chemosensitivity in these putative central respiratory chemoreceptor neurones.

CO2 dialysis in nucleus tractus solitarius region of rat increases ventilation in sleep and wakefulness.

To evaluate the function of widely distributed central chemoreceptors during sleep and wakefulness in the rat, we focally stimulate single chemoreceptor sites during naturally occurring sleep-wake cycles by microdialysis of artificial cerebrospinal fluid equilibrated with 25% CO2. In retrotrapezoid nucleus, this increased ventilation (tidal volume) by 24% only in wakefulness (Li A, Randall M, and Nattie E. J Appl Physiol 87: 910-919, 1999). In caudal medullary raphé, it increased ventilation (frequency) by 15-20% only in sleep (Nattie EE and Li A. J Appl Physiol 90: 1247-1257, 2001). Here, in nucleus tractus solitarius (NTS), focal acidification significantly increased ventilation by 11% in sleep and 7% in wakefulness rostrally (n = 5) and by 16% in sleep and 28% in wakefulness caudally (n = 5). The sleep-wake cycle was unaltered. Dialysis with 5% CO2 had no effect. Dialysis with 50% CO2 caudally did not further stimulate ventilation but did disrupt sleep. Central chemoreceptors in the NTS affect breathing in both sleep and wakefulness. The threshold for arousal in caudal NTS is greater than that for the stimulation of breathing.

Effect of ventilatory drive on carbon dioxide sensitivity below eupnea during sleep.

We determined the effects of changing ventilatory stimuli on the hypocapnia-induced apneic and hypopneic thresholds in sleeping dogs. End-tidal carbon dioxide pressure (PET(CO2)) was gradually reduced during non-rapid eye movement sleep by increasing tidal volume with pressure support mechanical ventilation, causing a reduction in diaphragm electromyogram amplitude until apnea/periodic breathing occurred. We used the reduction in PET(CO2) below spontaneous breathing required to produce apnea (DeltaPET(CO2)) as an index of the susceptibility to apnea. DeltaPET(CO2) was -5 mm Hg in control animals and changed in proportion to background ventilatory drive, increasing with metabolic acidosis (-6.7 mm Hg) and nonhypoxic peripheral chemoreceptor stimulation (almitrine; -5.9 mm Hg) and decreasing with metabolic alkalosis (-3.7 mm Hg). Hypoxia was the exception; DeltaPET(CO2) narrowed (-4.1 mm Hg) despite the accompanying hyperventilation. Thus, hyperventilation and hypocapnia, per se, widened the DeltaPET(CO2) thereby protecting against apnea and hypopnea, whereas reduced ventilatory drive and hypoventilation narrowed the DeltaPET(CO2) and increased the susceptibility to apnea. Hypoxia sensitized the ventilatory responsiveness to CO2 below eupnea and narrowed the DeltaPET(CO2); this effect of hypoxia was not attributable to an imbalance between peripheral and central chemoreceptor stimulation, per se. We conclude that the DeltaPET(CO2) and the ventilatory sensitivity to CO2 between eupnea and the apneic threshold are changeable in the face of variations in the magnitude, direction, and/or type of ventilatory stimulus, thereby altering the susceptibility for apnea, hypopnea, and periodic breathing in sleep.

Chemosensitive serotonergic neurons are closely associated with large medullary arteries.

We have previously shown that serotonergic neurons of the medulla are strongly stimulated by an increase in CO(2), suggesting that they are central respiratory chemoreceptors. Here we used confocal imaging and electron microscopy to show that neurons immunoreactive for tryptophan hydroxylase (TpOH) are tightly apposed to large arteries in the rat medulla. We used patch-clamp recordings from brain slices to confirm that neurons with this anatomical specialization are chemosensitive. Serotonergic neurons are ideally situated for sensing arterial blood CO(2), and may help maintain pH homeostasis via wide-ranging effects on brain function. The results reported here support a recent proposal that sudden infant death syndrome (SIDS) results from a developmental abnormality of medullary serotonergic neurons.

Temperature and pH/CO 2 modulate respiratory activity in the isolated brainstem of the bullfrog ( Rana catesbeiana)

The effects of temperature and pH/CO 2 were examined in isolated brainstem preparations from adult North American bullfrogs ( Rana catesbeiana). These experiments were undertaken to determine the effects of temperature on fictive breathing, central pH/CO 2 chemoreception, and to examine potential alphastat regulation of respiration in vitro. Adult bullfrog brainstem preparations were isolated, superfused with an artificial cerebrospinal fluid (aCSF) and respiratory-related neural activity was recorded from cranial nerves V, X and XII. In Series I experiments ( N=8), brainstem preparations were superfused with aCSF equilibrated with 2% CO 2 at temperatures ranging from 10 to 30 °C. Neural activity was present in all preparations in the temperature range of 15–25 °C, but was absent in most preparations when aCSF was at 10 or 30 °C. The absence of fictive breathing at high (30 °C) temperatures was transient since fictive breathing could be restored upon returning the preparation to 20 °C. In Series II experiments ( N=10), preparations were superfused with aCSF equilibrated with 0%, 2% and 5% CO 2 at temperatures of 15, 20 and 25 °C. Fictive breathing frequency ( f R) was significantly dependent upon aCSF pH at all three temperatures, with slopes ranging from −0.82 min −1 pH unit −1 (15 °C) to −3.3 min −1 pH unit −1 (20 °C). There was a significant difference in these slopes ( P<0.02), indicating that central chemoreceptor sensitivity increased over this temperature range. Fictive breathing frequency was significantly dependent upon the calculated alpha-imidazole (α Im) ionization ( P<0.05), consistent with the alphastat hypothesis for the effects of temperature on the regulation of ventilation. However, most of the variation in f R was not explained by α Im ( R 2=0.05), suggesting that other factors account for the regulation of fictive breathing in this preparation. The results indicate that the in vitro brainstem preparation of adult bullfrogs has a limited temperature range (15–25 °C) over which fictive breathing is consistently active. Although there is a close correspondence of ventilation in vitro and in vivo at low temperatures, these data suggest that, as temperature increases, changes in ventilation in the intact animal are likely to be more dependent upon peripheral feedback which assumes a greater integrative role with respect to chemoreceptor drive, respiratory frequency and tidal volume.

The effect of anesthetics on control of respiration

Ventilatory control in humans depends on complex mechanisms which aim to maintain a cellular CO2-, O2- and H(+)-homeostasis under physiological conditions. This regulation is based on chemical control which predominantly acts via peripheral chemoreceptors in the carotid bodies and central chemoreceptors in the ventral medulla of the brainstem on the one hand, and behavioural control on the other, by which it is possible to adapt respiration to conditions of daily living. The influence of anaesthesia and related conditions may depress respiration and have a sustained effect on ventilatory control. Perioperative respiratory depression remains a serious clinical problem in perioperative medicine. This review will give an overview of ventilatory control and discuss the most relevant responses, describe the effects of pain, anaesthetics and opioids on ventilatory control and their interaction. The current body of knowledge is put into perspective to identify patients at risk for perioperative respiratory depression.

Pathophysiologic Mechanisms Underlying Apnea of Prematurity

After completing this article, readers should be able to: 1. Characterize the unique features of neonatal respiratory control mechanisms. 2. Clarify how immature respiratory control contributes to neonatal apnea. 3. Identify the contribution of upper airway obstruction to episodes of mixed apnea. Idiopathic apnea of prematurity is a common disorder that requires therapeutic intervention to avoid potential morbidity in preterm infants who require neonatal intensive care. Severe, recurrent apneic episodes may lead to multiple investigations to rule out secondary disorders leading to apnea (see accompanying article in this issue). Although the incidence of neonatal apnea is inversely related to gestational age and is probably as high as 100% in the most immature preterm infants, the onset may be delayed by the presence of lung disease in the first days of life. Apnea of prematurity, especially if severe or persistent, has been associated with poor developmental outcome in school-age children, although a cause-and-effect relationship is difficult to establish. The definition of apnea varies among studies. Apnea of prematurity has been defined most widely as cessation of breathing in excess of 15 seconds’ duration, typically accompanied by desaturation and bradycardia. However, shorter episodes of apnea, and even periodic breathing, may be accompanied by bradycardia or hypoxemia. Prolonged desaturation episodes also have been reported in the absence of apnea or bradycardia, both in healthy preterm infants and in infants who have chronic lung disease. These episodes might represent obstructive apnea, hypoventilation, or possibly intrapulmonary right-to-left shunting, although episodes of desaturation and bradycardia almost always are preceded by apnea or hypoventilation. Apnea is classified traditionally into three categories based on the presence or absence of upper airway obstruction: central, obstructive, and mixed apneas. Central apnea is characterized by total cessation of inspiratory efforts with no evidence of obstruction. In obstructed apnea, the infant tries to breathe against …

The retrotrapezoid nucleus (RTN): local cytoarchitecture and afferent connections.

The retrotrapezoid nucleus (RTN) provides a source of tonic drive to respiratory neurons and is one of many sites for central chemoreception. Here we evaluate in the rat the local neuronal cytoarchitecture in the RTN histologically 2-4 h after neurobiotin injection and the afferent connections to the RTN 24 h after injection. Our neurobiotin injections often overlapped the RTN and the adjacent neurons of the parapyramidal region, so we group these two regions together in this study. The RTN is made up of small and medium sized neurons and has a low neuronal density compared to other nuclei. The organization of the RTN is reticular in nature and there are prominent small neurons at the ventral medullary border. Adjacent to the pyramids there are medium sized neurons with connections to the raphé pallidus. Major afferent connections include the regions of the dorsal and ventral respiratory groups, the medullary raphé, the contralateral parapyramidal and RTN regions, portions of the nucleus paragigantocellularis lateralis, and portions of the reticular fields. Other sources of input include the Kölliker-fuse nucleus, subceruleus, A5 region, and the paralemniscal zone.

Syndrome d’hypoventilation alvéolaire centrale et thrombose veineuse cérébrale : association fortuite ?

Central hypoventilation syndrome is defined as the failure of automatic control of breathing. Secondary central hypoventilation syndrome should distinguish from congenital central hypoventilation syndrome by brainstem abnormalities, place of respiratory control. Case reports. – We report two clinical cases characterized by late onset central hypoventilation syndrome (three years - six months, and five years old) : in the first case the diagnosis was made after general anesthesia and the second one presented with acute nocturnal comatose state. Neuroradiologic investigations showed bilateral cerebral sinus veinosus thrombosis without any brainstem lesions. Moreover these children had severe behavior disorders : psychomotor instability, alterations of social relations, language dysfunction, and neurocognitive deficit. This symptomatology seems independent from central hypoventilation syndrome and cerebral venous thombosis. Conclusion. – Late onset central hypoventilation syndrome may be associated with cerebral venous thrombosis. Ischemia of central chemoreceptors or integration of their informations could be one of mechanism.

Quantification of the response of rat medullary raphe neurones to independent changes in pHo and PCO2

The medullary raphe nuclei contain putative central respiratory chemoreceptor neurones that are highly sensitive to acidosis. To define the primary stimulus for chemosensitivity in these neurones, the response to hypercapnic acidosis was quantified and compared with the response to independent changes in PCO2 and extracellular pH (pHo). Neurones from the ventromedial medulla of neonatal rats (P0-P2) were dissociated and maintained in tissue culture for long enough to develop a mature response (up to 70 days). Perforated patch clamp recordings were used to record membrane potential and firing rate while changes were made in pHo, PCO2 and/or [NaHCO3]o from baseline values of 7.4, 5 % and 26 mm, respectively. Hypercapnic acidosis (PCO2 9 %; pHo 7.17) induced an increase in firing rate to 285 % of control in one subset of neurones (‘stimulated neurones’) and induced a decrease in firing rate to 21 % of control in a different subset of neurones (‘inhibited neurones’). Isocapnic acidosis (pHo 7.16; [NaHCO3]o 15 mm) induced an increase in firing rate of stimulated neurones to 309 % of control, and a decrease in firing rate of inhibited neurones to 38 % of control. In a different group of neurones, isohydric hypercapnia (9 % PCO2; [NaHCO3]o 40 mm) induced an increase in firing rate of stimulated neurones by the same amount (to 384 % of control) as in response to hypercapnic acidosis (to 327 % of control). Inhibited neurones also responded to isohydric hypercapnia in the same way as they did to hypercapnic acidosis. In Hepes-buffered solution, both types of neurone responded to changes in pHo in the same way as they responded to changes in pHo in bicarbonate-buffered Ringer solution. It has previously been shown that all acidosis-stimulated neurones in the medullary raphe are immunoreactive for tryptophan hydroxylase (TpOH-ir). Here it was found that TpOH-ir neurones in the medullary raphe were immunoreactive for carbonic anhydrase type II and type IV (CA II and CA IV). However, CA immunoreactivity was also common in neurones of the hypoglossal motor nucleus, inferior olive, hippocampus and cerebellum, indicating that its presence is not uniquely associated with chemosensitive neurones. In addition, under the conditions used here, acetazolamide (100 μm) did not have a significant effect on the response to hypercapnic acidosis. We conclude that chemosensitivity of raphe neurones can occur independently of changes in pHo, PCO2 or bicarbonate. The results suggest that a change in intracellular pH (pHi) may be the primary stimulus for chemosensitivity in these putative central respiratory chemoreceptor neurones.

Substance P-saporin lesion of neurons with NK1 receptors in one chemoreceptor site in rats decreases ventilation and chemosensitivity

All medullary central chemoreceptor sites contain neurokinin-1 receptor immunoreactivity (NK1R-ir). We ask if NK1R-ir neurons and processes are involved in chemoreception. At one site, the retrotrapezoid nucleus/parapyramidal region (RTN/Ppy), we injected a substance P-saporin conjugate (SP-SAP; 0.1 pmol in 100 nl) to kill NK1R-ir neurons specifically, or SAP alone as a control. We made measurements for 15 days after the injections in two groups of rats. In group 1, with unilateral injections made in the awake state via a pre-implanted guide cannula, we compared responses within rats using initial baseline data. In group 2, with bilateral injections made under anaesthesia at surgery, we compared responses between SP-SAP- and SAP-treated rats. SP-SAP treatment reduced the volume of the RTN/Ppy region that contained NK1R-ir neuronal somata and processes by 44 % (group 1) and by 47 and 40 % on each side, respectively (group 2). Ventilation (V̇E) and tidal volume (VT) were decreased during air breathing in sleep and wakefulness (group 2; P < 0.001; two-way ANOVA) and Pa,CO2 was increased (group 2; P < 0.05; Student's t test). When rats breathed an air mixture containing 7 % CO2 during sleep and wakefulness, V̇E and VT were lower (groups 1 and 2; P < 0.001; ANOVA) and the ΔV̇E in air containing 7 % CO2 compared to air was decreased by 28-30 % (group 1) and 17-22 % (group 2). SP-SAP-treated rats also slept less during air breathing. We conclude that neurons with NK1R-ir somata or processes in the RTN/Ppy region are either chemosensitive or they modulate chemosensitivity. They also provide a tonic drive to breathe and may affect arousal.

Evolution of central respiratory chemoreception: a new twist on an old story

Evolution of central respiratory chemosensitivity has been linked traditionally to the need for carbon dioxide regulation that accompanied the evolution of air breathing in terresterial animals. We examined the validity of this linkage by investigating the possibility of central chemoreception in air breathing fish that diverged from the amphibian lineage long before the appearance of terrestriality. We showed that the isolated brainstem preparation of the long nose gar ( Lepisosteus osseus) produces a putative motor pattern for lung ventilation, which is responsive to CO 2. These findings, together with more inferential evidence, suggest an association between air breathing and central chemosensitivity in aquatic animals that spans the major branches in vertebrate phylogeny. Furthermore, developmental observations in tadpoles suggest that the neural substrates for central chemoreception exist in proximity to that for rhythm generation. We postulate that a primitive ancestral CPG, sensitive to CO 2 is conserved and is evidenced in the intrinsic coupling of respiratory CPG and central chemoreception in modern tetrapods.

An alternative approach to the identification of respiratory central chemoreceptors in the brainstem

Central chemoreceptors (CCRs) play a crucial role in autonomic respiration. Although a variety of brainstem neurons are CO 2 sensitive, it remains to know which of them are the CCRs. In this article, we discuss a potential alternative approach that may allow an access to the CCRs. This approach is based on identification of specific molecules that are CO 2 or pH sensitive, exist in brainstem neurons, and regulate cellular excitability. Their molecular identity may provide another measure in addition to the electrophysiologic criteria to indicate the CCRs. The inward rectifier K + channels (Kir) seem to be some of the CO 2 sensing molecules, as they regulate membrane potential and cell excitability and are pH sensitive. Among homomeric Kirs, we have found that even the most sensitive Kir1.1 and Kir2.3 have pK∼6.8, suggesting that they may not be capable of detecting hypocapnia. We have studied their biophysical properties, and identified a number of amino acid residues and molecular motifs critical for the CO 2 sensing. By comparing all Kirs using the motifs, we found the same amino acid sequence in Kir5.1, and demonstrated the pH sensitivity in heteromeric Kir4.1 and Kir5.1 channels to be pK∼7.4. In current clamp, we show evidence that the Kir4.1–Kir5.1 can detect P CO 2 changes in either hypercapnic or hypocapnic direction. Our in-situ hybridization studies have indicated that they are coexpressed in brainstem cardio–respiratory nuclei. Thus, it is likely that the heteromeric Kir4.1–Kir5.1 contributes to the CO 2/pH sensitivity in these neurons. We believe that this line of research intended to identify CO 2 sensing molecules is an important addition to current studies on the CCRs.

Cytoarchitecture of central chemoreceptors in the mammalian ventral medulla

We reviewed the previous reports on the fine anatomy of the mammalian ventral medulla with special attention to the cytoarchitecture of the superficial chemosensitive regions to summarize what is known, what is not yet known, and what should be studied in the future. We also reviewed studies on anatomical relationship between neurons and vessels, and morphological studies on dendrites of respiratory or chemosensitive neurons. When we compared the morphological reports on the ventral and dorsal putative chemosensitive regions, similarities were found as follows. Chemosensitive cells were often found not only near the ventral surface but near the dorsal surface of the brainstem. Dendritic projection towards the surface was a common characteristic in the ventral and dorsal chemosensitive neurons. Morphological abnormality in the brainstem of sudden infant death syndrome victims was also summarized. On the basis of the previous reports we discussed the perspective on the future study on central chemoreception. Among various unanswered questions in central chemosensitivity studies, physiological significance of surface cells and surface extending dendrites is the most important topic, and must be thoroughly investigated.

Central chemosensitivity, sleep, and wakefulness

Neurons in many regions of the lower brain are chemosensitive in vitro. Focal acidification of these same and other regions in vivo can stimulate breathing indicating the presence of chemoreception. Why are there so many sites for central chemoreception? This review evaluates data obtained from unanesthetized rats at three central chemoreceptor sites, the retrotrapezoid nucleus (RTN), the medullary raphé, and the nucleus tractus solitarius (NTS) and extends ideas concerning two hypotheses, which were recently formulated (Nattie, E., 2000. Respir. Physiol. 122, 223–235). (1) The high overall sensitivity of the respiratory control system in the unanesthetized state to small increases in arterial CO 2 relies on an additive or greater effect of these multiple chemoreceptor sites. (2) Chemoreceptor sites can vary in effectiveness dependent on the state of arousal. These ideas fit into a more speculative and general hypothesis that central chemoreceptors are organized in a hierarchical manner as proposed for temperature sensing and thermoregulation (Satinoff, E., 1978. Science 201, 16–22). The presence of a number of chemosensitive sites with varying thresholds, sensitivity, and arousal dependence provides finely tuned control and stability for breathing.