Ventilatory Long-term Facilitation Research Articles

Sleep fragmentation impairs ventilatory long‐term facilitation via adenosine A1 receptors

Sleep fragmentation (SF), a primary feature of obstructive sleep apnoea (OSA), impairs hippocampal long-term potentiation and causes cognitive/attention deficits. However, its influence upon respiratory control has hardly been studied. This study examined the effect of SF on ventilatory long-term facilitation (LTF, a persistent augmentation of respiratory activity after episodic hypoxia) and the hypoxic ventilatory response (HVR), and investigated the role of adenosine A1 receptors in these SF effects in conscious adult male Sprague-Dawley rats. SF, confirmed by sleep architecture recordings, was achieved by periodic, forced locomotion in a rotating drum (30 s rotation/90 s stop for 24 h). LTF, elicited by five episodes of 5 min poikilocapnic hypoxia (10% O2) with 5 min intervals, was measured by plethysmography. Resting ventilation and metabolic rate were unchanged, HVR was reduced (150.6 +/- 3.5% versus 110.4 +/- 12.3%) and LTF was eliminated (22.6 +/- 0.5% versus -0.1 +/- 1.3%) shortly after 24 h SF. The SF-induced impairments were SF duration dependent, and completely reversible as HVR (< 24 h) and LTF (< 48 h) returned spontaneously to their pre-SF values. The SF-impaired HVR was improved (130.3 +/- 4.2%) and SF-eliminated LTF was restored (19.6 +/- 0.9%) by systemic injection of the adenosine A1 receptor antagonist 8-CPT (2.5 mg kg(-1)) approximately 30 min before LTF elicitation. Both HVR and LTF were also similarly impaired by 24 h total sleep deprivation or 24 h repeated cage tapping-induced SF, but not by a 24 h locomotion control protocol for SF. Collectively, these data suggest that: (1) 24 h SF impairs LTF and poikilocapnic HVR; (2) these impairments require A1 receptors; and (3) SF of OSA may exacerbate OSA via impaired ventilatory control mechanisms.

Formation and maintenance of ventilatory long-term facilitation require NMDA but not non-NMDA receptors in awake rats

N-methyl-d-aspartate (NMDA) receptor antagonism in the phrenic motonucleus area eliminates phrenic long-term facilitation (pLTF; a persistent augmentation of phrenic nerve activity after episodic hypoxia) in anesthetized rats. However, whether NMDA antagonism can eliminate ventilatory LTF (vLTF) in awake rats is unclear. The role of non-NMDA receptors in LTF is also unknown. Serotonin receptor antagonism before, but not after, episodic hypoxia eliminates pLTF, suggesting that serotonin receptors are required for induction, but not maintenance, of pLTF. However, because NMDA and non-NMDA ionotropic glutamate receptors are directly involved in mediating the inspiratory drive to phrenic, hypoglossal, and intercostal motoneurons, we hypothesized that these receptors are required for both formation and maintenance of vLTF. vLTF, induced by five episodes of 5-min poikilocapnic hypoxia (10% O(2)) with 5-min normoxia intervals, was measured with plethysmography in conscious adult male Sprague-Dawley rats. Either (+/-)-2-amino-5-phosphonovaleric acid (APV; NMDA antagonist, 1.5 mg/kg) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; non-NMDA antagonist, 10 mg/kg) was systemically (ip) injected approximately 30 min before hypoxia. APV was also injected immediately after or 20 min after episodic hypoxia in additional groups. As control, vehicle was similarly injected in each rat 1-2 days before. Regardless of being injected before or after episodic hypoxia, vehicle did not alter vLTF ( approximately 23%), whereas APV eliminated vLTF while having little effect on baseline ventilation or hypoxic ventilatory response. In contrast, CNQX enhanced vLTF ( approximately 34%) while decreasing baseline ventilation. Collectively, these results suggest that activation of NMDA but not non-NMDA receptors is necessary for formation and maintenance of vLTF in awake rats.

Orexinergic modulation of breathing across vigilance states

Basal respiration and respiratory reflex regulations are considerably different during the awake and sleep states. Tidal volume and respiratory frequency diminish during sleep, and hypoxic and hypercapnic ventilatory responses also decline during sleep. Reduced metabolic demand during sleep cannot completely explain these phenomena because PaCO2 increases during sleep. In this review, I will summarize our recent discovery of the possible contribution of orexin, a hypothalamic neuropeptide, to the vigilance state-dependent adjustment of central respiratory regulation. Orexin-deficient mice show an attenuated hypercapnic ventilatory response during the awake but not during the sleep period, whereas basal ventilation remained normal, irrespective of the vigilance state. Orexin supplementation remedied the defect, and the administration of an orexin receptor antagonist to wild-type mice mimicked the abnormality. Orexin-deficient mice also showed frequent sleep apneas and loss of repetitive intermittent hypoxia-induced ventilatory long-term facilitation. Hence, it is possible that the orexin system is one of the essential modulators required for coordinating the circuits controlling respiration and behavior.

Prenatal nicotine and respiratory long term facilitation in neonatal rats

Prenatal nicotine (PN) exposure alters the control of breathing in neonatal rats and the development of serotonergic neurons. Long term facilitation (LTF) is a long‐lasting, serotonin dependent increase in respiratory motor output that occurs in neonates and adults following intermittent hypoxia (IH). In ongoing studies, we are examining the impact of PN on ventilatory LTF in neonatal rats. Dams were implanted with an alzet miniosmotic pump to deliver nicotine (6 mg/kg per day) or saline during pregnancy. Barometric plethysmography was used to record ventilation in unanesthetized rat pups. Rats were exposed to a baseline period (21% O2) and then ten 5‐min bouts of hypoxia (5% O2) followed by a return to baseline conditions. Data recorded following IH were normalized to pre‐hypoxia baseline values. At post‐natal days 15–17, PN rats had a reduced inspiratory tidal volume (p=0.036 vs. saline, 2‐way RM ANOVA) and tended to have increased breathing frequency (p=0.215 vs. saline) over the 30 min period following IH. At 30 min post‐IH, tidal volume was larger than pre‐IH baseline values in saline (p=0.043) but not PN rats (p=0.767). Body weight was not different between PN (44±3 g) and saline rats (42±2 g). These preliminary results (n=4 per group) indicate that expression of LTF in neonatal rats is modulated by exposure to PN. Supported by the Florida Dept. of Health.

Chronic intermittent hypoxia reduces ventilatory long-term facilitation and enhances apnea frequency in newborn rats

Ventilatory long-term facilitation (LTF; defined as gradual increase of minute ventilation following repeated hypoxic exposures) is well described in adult mammals and is hypothesized to be a protective mechanism against apnea. In newborns, LTF is absent during the first postnatal days, but its precise developmental pattern is unknown. Accordingly, this study describes this pattern of postnatal development. Additionally, we tested the hypothesis that chronic intermittent hypoxia (CIH) from birth alters this development. LTF was estimated in vivo using whole body plethysmography by exposing rat pups at postnatal days 1, 4, and 10 (P1, P4, and P10) to 10 brief hypoxic cycles (nadir 5% O2) and respiratory recordings during the following 2 h (recovery, 21% O2). Under these conditions, ventilatory LTF (gradual increase of minute ventilation during recovery) was clearly expressed in P10 rats but not in P1 and P4. In a second series of experiments, rat pups were exposed to CIH during the first 10 postnatal days (6 brief cyclic exposures at 5% O2 every 6 min followed by 1 h under normoxia, 24 h a day). Compared with P10 control rats, CIH enhanced hypoxic ventilatory response (estimated during the hypoxic cycles) specifically in male rat pups. Ventilatory LTF was drastically reduced in P10 rats exposed to CIH, which was associated with higher apnea frequency during recovery. We conclude that CIH from birth enhances hypoxic chemoreflex and disrupts LTF development, thus likely contributing to increase apnea frequency.

Ventilatory long-term facilitation in mice can be observed during both sleep and wake periods and depends on orexin

Respiratory long-term facilitation (LTF) is a long-lasting (>1 h) augmentation of respiratory motor output that occurs even after cessation of hypoxic stimuli, is serotonin-dependent, and is thought to prevent sleep-disordered breathing such as sleep apnea. Raphe nuclei, which modulate several physiological functions through serotonin, receive dense projections from orexin-containing neurons in the hypothalamus. We examined possible contributions of orexin to ventilatory LTF by measuring respiration in freely moving prepro-orexin knockout mice (ORX-KO) and wild-type (WT) littermates before, during, and after exposure to intermittent hypoxia (IH; 5 x 5 min at 10% O2), sustained hypoxia (SH; 25 min at 10% O2), or sham stimulation. Respiratory data during quiet wakefulness (QW), slow wave sleep (SWS), and rapid-eye-movement sleep were separately calculated. Baseline ventilation before hypoxic stimulation and acute responses during stimulation did not differ between the ORX-KO and WT mice, although ventilation depended on vigilance state. Whereas the WT showed augmented minute ventilation (by 20.0 +/- 4.5% during QW and 26.5 +/- 5.3% during SWS; n = 8) for 2 h following IH, ORX-KO showed no significant increase (by -3.1 +/- 4.6% during QW and 0.3 +/- 5.2% during SWS; n = 8). Both genotypes showed no LTF after SH or sham stimulation. Sleep apnea indexes did not change following IH, even when LTF appeared in the WT mice. We conclude that LTF occurs during both sleep and wake periods, that orexin is necessary for eliciting LTF, and that LTF cannot prevent sleep apnea, at least in mice.

Ventilatory long-term facilitation in non-snoring subjects during NREM sleep

We hypothesized that very brief episodes of hypoxia (<1 min) would evoke long-term facilitation (LTF) in individuals free of inspiratory flow limitation (IFL). We studied 12 healthy participants who were self-reported non-snorers and confirmed the absence of IFL. We induced 15 brief episodes of hypoxia during non-REM sleep, reducing arterial oxygen saturation to 84-85%, followed by 1 min of room air. Ventilatory variables and resistance were measured during the control period, hypoxic trials, room air controls, and for 20 min following the last hypoxic episode. There was a significant increase in minute ventilation (108+/-1.3% of control, P < 0.05) and tidal volume (105+/-1.7% of control, P < 0.05) and a significant decrease in upper airway resistance (88+/-9.8% control, P < 0.05) during the recovery period. However, there were no significant changes in any variable during sham studies. We have shown for the first time that LTF can be elicited in sleeping humans free of IFL.

Thyroid status affects 5-HT2Areceptor modulation of breathing before, during, and following exposure of hamsters to acute intermittent hypoxia

The BIO 14.6 hamster (dystrophic), animal model of limb girdle muscular dystrophy, exhibits low plasma triiodothyronine levels, muscle weakness, and decreased breathing. After exposure to acute intermittent bouts of hypoxia, dystrophic hamsters depress ventilation relative to baseline resulting in ventilatory long-term depression (LTD). Control hamsters may increase ventilation relative to baseline resulting in ventilatory long-term facilitation (LTF). Serotonin (5-HT) receptors, especially the 5-HT(2A) subtype, are involved in the development of LTF. The purpose of this study was to evaluate the role of 5-HT(2A) receptors in ventilatory and metabolic responses before, during, and following intermittent hypoxia in eleven euthyroid, nine dystrophic, and eleven propylthiouracil (PTU)-induced hypothyroid male hamsters. Animals received subcutaneous injections of vehicle or 0.5 mg/kg MDL (5-HT(2A) receptor antagonist). Plethysmography was used to evaluate ventilatory responses of the three groups to air, five bouts of 5 min of 10% oxygen, each interspersed with 5 min of air, followed by 60 min of exposure to air. CO(2) production was measured using the flow-through method. Vehicle-treated dystrophic and PTU-treated hamsters exhibited LTD. MDL decreased body temperature in all groups. After MDL treatment, the euthyroid group exhibited LTD. MDL treatment in the dystrophic, but not in the PTU-treated hamsters, maintained tidal volume, but did not reverse LTD. CO(2) production was increased in the euthyroid group with MDL treatment. Thus, 5-HT(2A) receptors affect body temperature, ventilation, and metabolism in hamsters. The differential responses noted in this study may be in part dependent on thyroid hormone status.

In hamsters dopamine D 2 receptors affect ventilation during and following intermittent hypoxia

We tested the hypothesis that in golden Syrian hamsters ( Mesocricetus auratus) carotid body dopaminergic D 2 receptors modulate ventilation in air, during exposure to intermittent hypoxia (IH) and reoxygenation. Ventilation was evaluated using the barometric method and CO 2 production was determined using the flow through method. Hamsters ( n = 8) received either subcutaneous injections of vehicle, haloperidol (0.5 mg/kg) or domperidone (0.5 mg/kg). Ventilatory and metabolic variables were determined 30 min following injections, after each of 5 bouts of 5 min of 10% oxygen interspersed by normoxia (IH), and 15, 30, 45 and 60 min following IH when hamsters were exposed to air. Haloperidol, but not domperidone decreased body temperature in hamsters. Neither treatment affected CO 2 production. Vehicle-treated hamsters exhibited ventilatory long-term facilitation (VLTF) following IH. Haloperidol or domperidone decreased ventilation in air, during IH and eliminated VLTF due to changes in tidal volume and not frequency of breathing. Thus, in hamsters D 2 receptors are involved in control of body temperature and ventilation during and following IH.

Long-term facilitation of genioglossus activity is present in normal humans during NREM sleep.

Episodic hypoxia (EH) is followed by increased ventilatory motor output in the recovery period indicative of long-term facilitation (LTF). We hypothesized that episodic hypoxia evokes LTF of genioglossus (GG) muscle activity in humans during non-rapid eye movement sleep (NREM) sleep. We studied 12 normal non-flow limited humans during stable NREM sleep. We induced 10 brief (3 min) episodes of isocapnic hypoxia followed by 5 min of room air. Measurements were obtained during control, hypoxia, and at 5, 10, 20, 30 and 40 min of recovery, respectively, for minute ventilation (V(I)), supraglottic pressure (P(SG)), upper airway resistance (R(UA)) and phasic GG electromyogram (EMG(GG)). In addition, sham studies were conducted on room air. During hypoxia there was a significant increase in phasic EMG(GG) (202.7+/-24.1% of control, p<0.01) and in V (I) (123.0+/-3.3% of control, p<0.05); however, only phasic EMG(GG) demonstrated a significant persistent increase throughout the recovery. (198.9+/-30.9%, 203.6+/-29.9% and 205.4+/-26.4% of control, at 5, 10, and 20 min of recovery, respectively, p<0.01). In multivariate regression analysis, age and phasic EMG(GG) activity during hypoxia were significant predictors of EMG(GG) at recovery 20 min. No significant changes in any of the measured parameters were noted during sham studies. (1) EH elicits LTF of GG in normal non-flow limited humans during NREM sleep, without concomitant ventilatory or mechanical LTF. (2) GG activity during the recovery period correlates with the magnitude of GG activation during hypoxia, and inversely with age.

Long-term facilitation of breathing is absent after episodes of hypercapnic hypoxia in awake humans

Despite the failure by many previous investigators to demonstrate a long-term facilitation of breathing following episodes of hypoxia in awake humans, we attempted to produce it using a pattern of hypercapnic hypoxic episodes similar to that experienced by obstructive sleep apnoea patients, reasoning that if long-term facilitation was relevant to these patients then it is appropriate to test the effectiveness of such episodes. Ten subjects drawn from the University student population were instrumented to measure ventilation, heart rate and end-tidal PCO2 and PO2 breath-by-breath while seated in a comfortable reclining chair. After an initial resting period breathing room air they experienced fifteen, 30-s episodes breathing 6% O2 and 5% CO2 separated by 90 s of breathing air. We examined the measured variables for an hour after the episodes but found no trends toward an increase in ventilation or decrease in end-tidal PCO2 that would indicate the presence of a long-term facilitation. We therefore concluded that long-term facilitation of ventilation was not demonstrated in awake humans using this pattern of stimuli.

Long-term facilitation of ventilation and genioglossus muscle activity is evident in the presence of elevated levels of carbon dioxide in awake humans

We hypothesized that long-term facilitation (LTF) of minute ventilation and peak genioglossus muscle activity manifests itself in awake healthy humans when carbon dioxide is sustained at elevated levels. Eleven subjects completed two trials. During trial 1, baseline carbon dioxide levels were maintained during and after exposure to eight 4-min episodes of hypoxia. During trial 2, carbon dioxide was sustained 5 mmHg above baseline levels during exposure to episodic hypoxia. Seven subjects were exposed to sustained elevated levels of carbon dioxide in the absence of episodic hypoxia, which served as a control experiment. Minute ventilation was measured during trial 1, trial 2, and the control experiment. Peak genioglossus muscle activity was measured during trial 2. Minute ventilation during the recovery period of trial 1 was similar to baseline (9.3 +/- 0.5 vs. 9.2 +/- 0.7 l/min). Likewise, minute ventilation remained unchanged during the control experiment (beginning vs. end of control experiment, 14.4 +/- 1.7 vs. 14.7 +/- 1.4 l/min). In contrast, minute ventilation and peak genioglossus muscle activity during the recovery period of trial 2 was greater than baseline (minute ventilation: 28.4 +/- 1.7 vs. 19.6 +/- 1.0 l/min, P < 0.001; peak genioglossus activity: 1.6 +/- 0.3 vs. 1.0 fraction of baseline, P < 0.001). We conclude that exposure to episodic hypoxia is necessary to induce LTF of minute ventilation and peak genioglossus muscle activity and that LTF is only evident in awake humans in the presence of sustained elevated levels of carbon dioxide.

Intermittent but not sustained hypercapnic hypoxia induces ventilatory long‐term facilitation in awake rats

Obstructive sleep apnea (OSA) patients experience hypercapnia as well as hypoxemia during their recurrent apnea events. However, respiratory long-term facilitation (LTF), which may play a role in stabilizing upper airway patency in OSA patients as some postulated, has so far been studied almost exclusively using intermittent hypoxia alone. In anesthetized rats, intermittent, but not sustained, hypoxia induces LTF, whereas neither intermittent nor sustained hypercapnia induces LTF. The aim of this study was to systematically examine the effectiveness of either intermittent or sustained exposure to hypoxia and/or hypercapnia in inducing ventilatory LTF in awake rats. Thus adult male Sprague-Dawley rats were exposed to 9 different protocols: 10 episodes of 2-min hypercapnic hypoxia (HH: 5% CO2 plus 10% O2), hypoxia (HX: 10% O2) or hypercapnia (HC: 5% CO2) with 3-min normoxia intervals, and sustained HH, HX or HC with 47-min (entire protocols) or 20-min (accumulative stimuli) duration. LTF, measured by plethysmography, was successfully induced by intermittent HH (23% above baseline) and intermittent HX (20%), but not by intermittent HC (3%) or any of the sustained protocols (HH: 2% and 3%, HX: 2% and −1%, HC: −5% and −3%). These results indicate that intermittent HH, which more closely mimics sleep apneas compared with intermittent HX, induces a similar, if not greater, ventilatory LTF in awake rats. These data also confirm in awake rats that neither intermittent nor sustained HC induces LTF, and that the pattern of intermittent stimuli is essential since sustained HH or HX fails to induce LTF even with relatively longer durations. (Supported by NIH HL64912)

Ten hypoxic episodes induce similar ventilatory long‐term facilitation in Lewis and Brown Norway rats

Acute intermittent hypoxia (3 to 5 episodes) elicits a form of respiratory plasticity known as long-term facilitation (LTF), a prolonged increase in phrenic, hypoglossal, and/or ventilatory output after the final hypoxic episode. Preconditioning rats for 7 days with daily acute intermittent hypoxia (dAIH; 10, 5-min episodes, 11±0.5% O2) enhances LTF in Brown Norway rats, an example of respiratory metaplasticity (Wilkerson et al., FASEB J. 2005). To begin exploring cellular/synaptic mechanisms of enhanced LTF following dAIH, we performed experiments to determine the magnitude of ventilatory LTF (vLTF) following the first day of dAIH, and harvested the tissues for protein and mRNA analyses. Here we report the ventilatory data for Lewis and Brown Norway rats, two inbred rat strains with differential phrenic LTF expression. Ventilation was measured in adult, male Lewis (Colony 202B, n=31) and Brown Norway (Colony 217, n=32) rats via whole body plethysmography before, during and 60 min following 10, 5-min hypoxic episodes (10.5%; ie. 1 day of dAIH) or in normoxic time controls. Significant vLTF was observed in both rat strains relative to normoxic time controls at 25–30 (p<0.05), but not 55–60 min post-hypoxia; vLTF was expressed primarily as increased tidal volume with variable frequency effects. Strain differences in vLTF were not observed. Thus, one day of dAIH elicits similar vLTF in the two inbred rat strains, although the magnitude and duration of this effect is limited. Supported by NIH HL65383, HL89209, and HL69064.

Ventilatory long-term facilitation following intermittent hypoxia is state-dependent in rats

Ventilatory long-term facilitation following intermittent hypoxia is state-dependent in rats

The ventilatory response to carbon dioxide and sustained hypoxia is enhanced after episodic hypoxia in OSA patients

Our primary hypothesis was that the acute ventilatory response to carbon dioxide in the presence of sustained hypoxia {VRCO2 (hypoxia)} or hyperoxia {VRCO2 (hyperoxia)} would increase in subjects with obstructive sleep apnea (OSA) after exposure to episodic hypoxia. Secondarily, we hypothesized that chronic (i.e. years) exposure to episodic hypoxia, a hallmark of OSA, would facilitate persistent augmentation of respiratory activity (i.e. long-term facilitation) after acute (i.e. minutes) exposure to episodic hypoxia. Nine healthy males with OSA that were healthy otherwise completed a series of rebreathing trials before and after exposure to eight 4 min episodes of hypoxia. On a separate occasion, the rebreathing trials were repeated before and after exposure to atmospheric air for a duration equivalent to the episodic hypoxia protocol (i.e. sham episodic hypoxia). During the rebreathing trials, subjects initially hyperventilated to reduce the partial pressure of carbon dioxide (P(ET)CO2) below 25 Torr. Subjects then rebreathed from a bag containing a normocapnic (42 Torr), low (50 Torr) or high oxygen gas mixture (140 Torr). During the trials, P(ET)CO2 increased while the selected level of oxygen was maintained. The point at which ventilation began to rise in a linear fashion as P(ET)CO2 increased was the ventilatory threshold. The ventilatory response below and above the threshold was determined. The results showed that the VRCO2 (hypoxia) and the VRCO2 (hyperoxia) was increased after exposure to episodic hypoxia {VRCO2 (hypoxia): 7.9 +/- 1.3 versus 10.5 +/- 1.3, VRCO2 (hyperoxia): 5.9 +/- 1.1 versus 6.7 +/- 1.1 L/min/Torr}. However, only the increase in the VRCO2 (hypoxia) after episodic hypoxia was greater than the increase measured after exposure to sham episodic hypoxia. Long-term facilitation of ventilation, tidal volume and breathing frequency was not evident after episodic hypoxia. We conclude that the VRCO2 (hypoxia) is enhanced after exposure to acute episodic hypoxia and that enhancement of the VRCO2 (hypoxia) occurs even though long-term facilitation is not evident.

Changes in ventilatory adaptations associated with long-term intermittent hypoxia across the age spectrum in the rat

Intermittent hypoxia (IH) induces alterations in respiratory control that reflect various types of ventilatory plasticity. In freely behaving rats, acute exposure to IH elicits enhancements in normoxic minute ventilation (VE), termed ventilatory long-term facilitation. Exposure to longer time periods of IH induces unique ventilatory adaptations to intermittent hypoxia (VAIH). We hypothesized that long-term IH-induced ventilatory plasticity may be developmentally regulated and thus, IH exposures at progressively later post-natal ages may elicit differential effects on the magnitude of VAIH. To examine this issue, male Sprague-Dawley rats were exposed to 30 continuous days of IH beginning at post-natal ages 1, 10, 30, 60, 180, 360, and 540 days. Control animals were exposed to normoxic conditions with room air. Normoxic VE was significantly higher in IH-exposed rats (p < 0.01) except for the group in which IH was initiated at post-natal age 540 days (p = NS). The magnitude of VAIH was greatest in rats exposed in the immediate post-natal period and gradually diminished with advancing post-natal age. Enhanced normoxic VE was due to significant contributions from both frequency (p < 0.01) and tidal volume (p < 0.01), and could not be accounted for by changes in metabolic rate. We conclude that the magnitude of IH-induced ventilatory plasticity is age-dependent with progressive declines becoming apparent with advancing post-natal age.

Ventilatory long-term facilitation is greater in 1- vs. 2-mo-old awake rats

Respiratory long-term facilitation (LTF) declines in middle-aged vs. adult male rats. Chronic intermittent hypoxia (CIH; 5 min 11-12% O2/5 min air, 12 h/night, 7 nights) enhances LTF in adult rats. However, LTF in immature rats and the effect of early CIH are unevaluated. The present study compared LTF in 1- and 2-mo-old rats and examined the effect of neonatal CIH (initiated at 2 days after birth) on the LTF. Ventilatory LTF, elicited by 5 (protocol 1) or 10 (protocol 2) episodes of poikilocapnic hypoxia (5 min 12% O2/5 min air), was measured twice by plethysmography on the same male conscious rat when it was 1 and 2 mo old. In untreated (without CIH) rats, both resting ventilation (54.7 +/- 0.6 vs. 43.0 +/- 0.2 ml.100 g(-1).min(-1)) and hypoxic ventilatory response (131 +/- 4 vs. 66 +/- 3% above baseline) were greater in 1- vs. 2-mo-old rats. Protocol 1 elicited LTF in 1-mo-old (12.5 +/- 1.0% above baseline) but not 2-mo-old rats. Protocol 2 elicited a greater LTF in 1-mo-old (24.3 +/- 0.8%) vs. 2-mo-old rats (18.2 +/- 0.5%). In CIH-treated rats, protocol 1 also elicited LTF in 1-mo-old (13.1 +/- 1.5%) but not 2-mo-old rats. Protocol 2 elicited LTF in both age groups, but LTF was enhanced by the CIH only in 1-mo-old rats (28.8 +/- 0.9%). These results suggest that ventilatory LTF and hypoxic ventilatory response are greater in male rats shortly before their sexual maturity and that the neonatal CIH somewhat enhances ventilatory LTF approximately 3 wk after CIH, but this enhancement does not last to adulthood.

Serotonin receptor subtypes required for ventilatory long-term facilitation and its enhancement after chronic intermittent hypoxia in awake rats.

Respiratory long-term facilitation (LTF), a serotonin-dependent, persistent augmentation of respiratory activity after episodic hypoxia, is enhanced by pretreatment of chronic intermittent hypoxia (CIH; 5 min 11-12% O2-5 min air, 12 h/night for 7 nights). The present study examined the effects of methysergide (serotonin 5-HT1,2,5,6,7 receptor antagonist), ketanserin (5-HT2 antagonist), or clozapine (5-HT2,6,7 antagonist) on both ventilatory LTF and the CIH effect on ventilatory LTF in conscious male adult rats to determine which specific receptor subtype(s) is involved. In untreated rats (i.e., animals not exposed to CIH), LTF, induced by five episodes of 5-min poikilocapnic hypoxia (10% O2) separated by 5-min normoxic intervals, was measured twice by plethysmography. Thus the measurement was conducted 1-2 days before (as control) and approximately 1 h after systemic injection of methysergide (1 mg/kg ip), ketanserin (1 mg/kg), or clozapine (1.5 mg/kg). Resting ventilation, metabolic rate, and hypoxic ventilatory response (HVR) were unchanged, but LTF ( approximately 18% above baseline) was eliminated by each drug. In CIH-treated rats, LTF was also measured twice, before and approximately 8 h after CIH. Vehicle, methysergide, ketanserin, or clozapine was injected approximately 1 h before the second measurement. Neither resting ventilation nor metabolic rate was changed after CIH and/or any drug. HVR was unchanged after methysergide and ketanserin but reduced in four of seven clozapine rats. The CIH-enhanced LTF ( approximately 28%) was abolished by methysergide and clozapine but only attenuated by ketanserin (to approximately 10%). Collectively, these data suggest that ventilatory LTF requires 5-HT2 receptors and that the CIH effect on LTF requires non-5-HT2 serotonin receptors, probably 5-HT6 and/or 5-HT7 subtype(s).

Chronic intermittent hypoxia enhances ventilatory long-term facilitation in awake rats.

This study examined the effect of chronic intermittent hypoxia (CIH: 5 min 11-12% O2/5 min air, 12 h/night, 7 nights) on ventilatory long-term facilitation (LTF) and determined the persistence period of this CIH effect in awake rats. LTF, elicited by 5 or 10 episodes of 5 min 12% O2, was measured four times in the same Sprague-Dawley rats by plethysmography, before and 8 h, 3 days, and 7 days after CIH treatment. Resting ventilation was unchanged after CIH. Five episodes of 12% O2 did not initially elicit LTF but elicited LTF (23.5 +/- 1.4% above baseline) 8 h after CIH, which partially remained at 3 days (11.4 +/- 2.2%, P < 0.05) and disappeared at 7 days. Ten episodes initially elicited LTF (17.7 +/- 1.1%, 45-min duration) and elicited an enhanced LTF (29.1 +/- 1.5%, 75 min) 8 h after CIH. These results demonstrated that CIH enhanced ventilatory LTF in conscious, freely behaving rats in two ways: 1) a previously ineffective protocol induced LTF; and 2) LTF magnitude was increased and LTF duration prolonged, and this CIH effect on LTF persisted for at least 3 days.