Discharge patterns of somatosensitive neurons in the nucleus tractus solitarius of the cat.

Encoding of sensory information by nucleus of the solitary tract (NTS) neurons is incompletely understood. Using extracellular single-unit recording in alpha-chloralose-urethane anesthetized cats, we have examined the discharge characteristics of NTS neurons to activation of somatic Adelta and C fiber afferents by skeletal muscle contraction evoked by electrical stimulation of lower lumbar/upper sacral ventral roots. Generally, somatic afferent stimulation evoked two distinct firing patterns. The first population (36/43 cells) increased their firing rate to brief somatic stimuli. A subset (21/27 cells) exhibited a rapid decay of their firing rate during sustained somatic stimulation. Peak instantaneous firing frequency (F(p)) increased proportionally with the intensity of somatic stimulation (105+/-4 vs. 119+/-4 vs. 139+/-4 Hz, 10, 20 and 40 Hz, respectively, P<0.0001), whereas steady-state firing frequency (F(ss)) was not altered (25+/-2 vs. 27+/-2 vs. 27+/-2 Hz, 10, 20 and 40 Hz, respectively, P=0.72). Two indices were derived to quantify the decay properties. The decay rate constant (obtained from exponential curve fitting) was not altered by stimulation frequency (461+/-10 vs. 442+/-14 vs. 429+/-26 ms, 10, 20 and 40 Hz, respectively, P=0.415), nor was the decay index (derived to express the percent reduction in firing rate with respect to the initial peak firing rate; 76+/-2 vs. 77+/-2 vs. 81+/-2%, 10, 20 and 40 Hz, respectively, P=0.187). In contrast, the second population (seven of 43 cells) decreased their firing rate to stimulation. Of the NTS neurons tested for barosensitivity (29/36), none responded to pressure stimulation. These results have identified a population of somatosensitive NTS neurons that exhibit rapid firing rate decay properties during sustained stimulation. However, this population could faithfully encode phasic excitation during rhythmic somatosensory input. These results are discussed in relation to the role of somatosensory input on baroreflex function.

Spontaneously hypertensive rats exhibit altered cardiovascular and neuronal responses to muscle contraction.

We examined the cardiovascular and ventrolateral medullary neuronal responses to muscle contraction in the spontaneously hypertensive rat (SHR) and normotensive Wistar-Kyoto rat (WKY) control. Cardiovascular, respiratory and ventrolateral medullary neuronal responses to muscle contraction evoked by tibial nerve stimulation were recorded. SHRs exhibited significantly larger drops in arterial pressure compared to WKYs in response to muscle contraction (P < 0.05). Basal ventrolateral medulla neuronal discharge rates were similar between the SHR and the WKY groups. A majority of neurons recorded responded to muscle contraction in both the WKY (77 %; n = 53) and the SHR groups (68 %; n = 62). There was no difference in the percentage of neurons that responded with an increase (approximately 60 %) or decrease (approximately 40 %) in firing rate between hypertensive and normotensive rats. Pulse wave-triggered averaging techniques showed that most neurons that responded to muscle contraction also possessed a basal firing rhythm temporally related to the cardiac cycle (85 % in WKYs, 83 % in SHRs). However, decreases in neuronal firing rates in response to muscle contraction were significantly greater in SHRs than WKYs. Therefore, we conclude that muscle contraction unmasks a hyperexcitability of neurons in the ventrolateral medulla of SHRs that parallels the heightened blood pressure responses.

Chronic exercise increases GAD gene expression in the caudal hypothalamus of spontaneously hypertensive rats.

Previous studies have suggested that a gamma-amino-butyric acid (GABA) deficit in the caudal hypothalamus (CH) of the spontaneously hypertensive rat (SHR) contributes to elevated levels of arterial pressure. The purpose of this study was to examine if SHR that underwent exercise training demonstrated a blunted development of hypertension and greater levels of glutamic acid decarboxylase (GAD) mRNA transcripts in the caudal hypothalamus. SHR were randomly paired and assigned to either a trained group (T; n=9) or a non-trained control group (NT; n=9). Trained animals were exercised for 10 weeks on a motorized treadmill while NT animals concurrently rested on a mock-treadmill. Following the 10-week training period, Northern blot analyses of mRNA for both the 65-kDa (GAD(65)) and 67-kDa (GAD(67)) isoforms of GAD were performed on tissue from caudal hypothalamic and cerebellar control brain regions. Exercise training simultaneously blunted the developmental rise in blood pressure in SHR (Delta59+/-9 mmHg in trained versus Delta77+/-9 mmHg in non-trained; P<0.03) and increased both GAD(65) (147+/-44%) and GAD(67) (162+/-77%) mRNA transcript levels in the CH (P<0.05). In contrast, no difference was detected in GAD mRNA levels in the cerebellum between T and NT SHR. These findings are consistent with our previous functional studies and demonstrate that exercise can significantly and specifically upregulate GAD gene transcript levels in the caudal hypothalamus of hypertensive rats.

Chronic exercise alters caudal hypothalamic regulation of the cardiovascular system in hypertensive rats.

Previous studies have documented a deficit in the GABA neurotransmitter system within the caudal hypothalamus (CH) of spontaneously hypertensive rats (SHR). The reduction in inhibitory influence on this cardiovascular excitatory brain region is associated with an increased neuronal activity and resting blood pressure. The purpose of this study was to determine if chronic treadmill and wheel-running activities alter the ability of the CH to regulate cardiovascular function. SHR were exercised on a treadmill (5 times/wk) at moderate intensity or allowed free access to running wheels (7 days/wk) for a period of 10 wk. Resting blood pressures were obtained before and after the exercise training periods. After the exercise period, rats were anesthetized and microinjection experiments were performed. Treadmill-trained SHR exhibited a significantly blunted developmental rise in resting blood pressure after 10 wk of exercise. A similar yet less marked effect was observed in wheel-run rats. Microinjection of the GABA synthesis inhibitor 3-mercaptopropionic acid (3-MP) into the CH of nonexercised SHR did not produce any change in arterial pressure. In contrast, microinjection of 3-MP into the CH produced significant increases in blood pressure and heart rate in exercised SHR. These results demonstrate that exercise training can alter CH cardiovascular regulation in hypertensive rats and therefore may play a role in increasing cardiovascular health.

Reflex cardiovascular responses originating in exercising muscles of mice.

The cardiovascular responses induced by exercise are initiated by two primary mechanisms: central command and reflexes originating in exercising muscles. Although our understanding of cardiovascular responses to exercise in mice is progressing, a murine model of cardiovascular responses to muscle contraction has not been developed. Therefore, the purpose of this study was to characterize the cardiovascular responses to muscular contraction in anesthetized mice. The results of this study indicate that mice demonstrate significant increases in blood pressure (13.8 +/- 1.9 mmHg) and heart rate (33.5 +/- 11.9 beats/min) to muscle contraction in a contraction-intensity-dependent manner. Mice also demonstrate 23.1 +/- 3.5, 20.9 +/- 4.0, 21.7 +/- 2.6, and 25.8 +/- 3.0 mmHg increases in blood pressure to direct stimulation of tibial, peroneal, sural, and sciatic hindlimb somatic nerves, respectively. Systemic hypoxia (10% O(2)-90% N(2)) elicits increases in blood pressure (11.7 +/- 2.6 mmHg) and heart rate (42.7 +/- 13.9 beats/min), while increasing arterial pressure with phenylephrine decreases heart rate in a dose-dependent manner. The results from this study demonstrate the feasibility of using mice to study neural regulation of cardiovascular function during a variety of autonomic stimuli, including exercise-related drives such as muscle contraction.

Cardiopulmonary function in rats with lung hemorrhage induced by pulsed ultrasound exposure.

OBJECTIVE: To assess cardiopulmonary function in rats exposed to pulsed ultrasound using superthreshold exposure conditions known to produce significant lung hemorrhage. METHODS: In 1 group of 9 anesthetized Sprague-Dawley rats, 5 foci of ultrasound-induced hemorrhage were produced in the left lung of each rat. In a second group of 6 rats, 5 foci of ultrasound-induced hemorrhage were produced in the left and right lungs of each rat. Each lesion was induced using superthreshold pulsed ultrasound exposure conditions (3.1-MHz center frequency, 1.7-kHz pulse repetition frequency, 1.3-micro-second pulse duration, 60-second exposure duration, 39-MPa in situ peak compressional pressure, and 17-MPa in situ peak rarefactional pressure). After exposure, the lungs were fixed in formalin and assessed histologically. The total lesion volume was calculated for each lesion in each lung lobe. Measurements of cardiopulmonary function included assessment of pulsatile arterial pressure, heart rate, end-tidal carbon dioxide, respiratory rate, and arterial blood gases (PCO2 and PO2). Functional data were quantified before (baseline) and 30 minutes after exposure to ultrasound. RESULTS: In the 9 rats that had lesions in only the left lung, the mean (SEM) lesion volume was 97 (13) mm3 and represented about 3.4% of the total lung volume. In the 6 rats that had lesions in both the left and right lungs, the left, right, and total mean lesion volumes, respectively, were 102 (16), 114 (11), and 216 (18) mm3 and represented about 3.7%, 4.2%, and 7.9% of the total lung volume. There were no statistically significant differences in cardiopulmonary measurements between baseline values and values obtained after exposure to ultrasound in the 9 rats exposed on the left lung only. The 6 rats exposed bilaterally had statistically significant differences in arterial pressure (134 +/- 4 versus 113 +/- 9 mm Hg; P= .047) and arterial PO2 (70 +/- 5 versus 58 +/- 4 mm Hg; P = .024) between baseline values and values obtained after exposure to ultrasound. CONCLUSIONS: The severity of ultrasound-induced lesions produced in 1 lung did not affect measurements of cardiopulmonary function because of the functional respiratory reserve in the unexposed lung. However, when both the left and right lungs had ultrasound-induced lesions, the functional respiratory reserve was decreased to a point at which rats were unable to maintain systemic arterial pressure or resting levels of arterial PO2.

Biophysical characterization of rat caudal hypothalamic neurons: calcium channel contribution to excitability.

Neurons in the caudal hypothalamus (CH) are responsible for the modulation of various processes including respiratory and cardiovascular output. Previous results from this and other laboratories have demonstrated in vivo that these neurons have firing rhythms matched to the respiratory and cardiovascular cycles. The goal of the present study was to characterize the biophysical properties of neurons in the CH with particular emphasis in those properties responsible for rhythmic firing behavior. Whole cell, patch-clamped CH neurons displayed a resting membrane potential of -58.0 +/- 1.1 mV and an input resistance of 319.3 +/- 16.6 MOmega when recorded in current-clamp mode in an in vitro brain slice preparation. A large proportion of these neurons displayed postinhibitory rebound (PIR) that was dependent on the duration and magnitude of hyperpolarizing current as well as the resting membrane potential of the cell. Furthermore these neurons discharged tonically in response to a depolarizing current pulse at a depolarized resting membrane potential (more positive than -65 mV) but switched to a rapid burst of firing to the same stimulus when the resting membrane potential was lowered. The PIR observed in these neurons was calcium dependent as demonstrated by the ability to block its amplitude by perfusion of Ca(2+)-free bath solution or by application of Ni(2+) (0.3-0.5 mM) or nifedipine (10 microM). These properties suggest that low-voltage-activated (LVA) calcium current is involved in the PIR and bursting firing of these CH neurons. In addition, high-voltage-activated calcium responses were detected after blockade of outward potassium current or in Ba(2+)-replacement solution. In addition, almost all of the CH neurons studied showed spike frequency adaptation that was decreased following Ca(2+) removal, indicating the involvement of Ca(2+)-dependent K(+) current (I(K,Ca)) in these cells. In conclusion, CH neurons have at least two different types of calcium currents that contribute to their excitability; the dominant current is the LVA or T-type. This LVA current appears to play a significant role in the bursting characteristics that may underlie the rhythmic firing of CH neurons.

Hypoxic augmentation of fast-inactivating and persistent sodium currents in rat caudal hypothalamic neurons.

Previous work from this laboratory has indicated that TTX-sensitive sodium channels are involved in the hypoxia-induced inward current response of caudal hypothalamic neurons. Since this inward current underlies the depolarization and increased firing frequency observed in these cells during hypoxia, the present study utilized more detailed biophysical methods to specifically determine which sodium currents are responsible for this hypoxic activation. Caudal hypothalamic neurons from approximately 3-wk-old Sprague-Dawley rats were acutely dissociated and patch-clamped in the voltage-clamp mode to obtain recordings from fast-inactivating and persistent (noninactivating) whole cell sodium currents. Using computer-generated activation and inactivation voltage protocols, rapidly inactivating sodium currents were analyzed during normal conditions and during a brief (3-6 min) period of severe hypoxia. In addition, voltage-ramp and extended-voltage-activation protocols were used to analyze persistent sodium currents during normal conditions and during hypoxia. A polarographic oxygen electrode determined that the level of oxygen in this preparation quickly dropped to 10 Torr within 2 min of initiation of hypoxia and stabilized at <0.5 Torr within 4 min. During hypoxia, the peak fast-inactivating sodium current was significantly increased throughout the entire activation range, and both the activation and inactivation values (V(1/2)) were negatively shifted. Furthermore both the voltage-ramp and extended-activation protocols demonstrated a significant increase in the persistent sodium current during hypoxia when compared with normoxia. These results demonstrate that both rapidly inactivating and persistent sodium currents are significantly enhanced by a brief hypoxic stimulus. Furthermore the hypoxic-induced increase in these currents most likely is the primary mechanism for the depolarization and increased firing frequency observed in caudal hypothalamic neurons during hypoxia. Since these neurons are important in modulating cardiorespiratory activity, the oxygen responsiveness of these sodium currents may play a significant role in the centrally mediated cardiorespiratory response to hypoxia.

Hypothalamus, hypertension, and exercise.

The hypothalamus is a well-known autonomic regulatory region of the brain involved in integrating several behaviors as well as cardiorespiratory activity. Our laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses associated with exercise. In addition, other findings from this laboratory and others have implicated alterations in this same brain region in spontaneously hypertensive rats as contributing factors of the elevated levels of arterial pressure in hypertension. Several studies have revealed a gamma-amino-butyric acid (GABAergic) deficiency in the caudal hypothalamus of spontaneously hypertensive rats that contributes to the tonic disinhibition and overactivity of this pressor region. Because chronic exercise is able to increase cardiovascular health in the hypertensive rat, we hypothesized that exercise-induced caudal hypothalamic plasticity partially underlies the beneficial effects of physical activity. In this review we discuss initial findings from this lab that support this hypothesis. Our experiments demonstrate that chronic exercise alters gene expression and neuronal activity in the caudal hypothalamus of the spontaneously hypertensive rat. These findings describe a potential mechanism by which chronic exercise lowers blood pressure in the hypertensive individual.

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

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

In vitro responses of neurons in the periaqueductal gray to hypoxia and hypercapnia.

Hypoxia-sensitive neurons in the caudal hypothalamus (CH) have been shown to project to the periaqueductal gray (PAG) which, in turn, sends descending projections to an area of the ventrolateral medulla (VLM) containing neurons inherently excited by hypoxia. The purpose of this study was to determine if neurons in the PAG are excited by hypoxia or hypercapnia in an in vitro environment. Extracellular responses to hypoxia and hypercapnia of neurons located throughout the PAG were recorded in a rat brain slice (400-500 microm thick) preparation. Hypoxic (10% O(2)/5% CO(2)/85% N(2)) and hypercapnic (7% CO(2)/93% O(2)) stimuli were delivered to the tissue through gas bubbled into the brain slice chamber. A majority (39 of 53) of the neurons tested responded to hypoxia. Of these neurons, 92% responded to hypoxia with an increase in firing rate. Neurons in the dorsolateral/lateral regions increased firing rates to a greater extent than neurons located in ventrolateral regions. All neurons tested (n=6) also responded to hypoxia after perfusion of the tissue with a low Ca(2+)/high Mg(2+) solution to block classic synaptic transmission. Only a small proportion (7/33) of neurons tested responded to hypercapnia. These findings indicate that neurons in the periaqueductal gray region of the brain have an inherent responsiveness to hypoxia and, thus, may contribute to the overall coordination of cardiorespiratory responses to systemic hypoxia.

Developmental aspects and mechanisms of rat caudal hypothalamic neuronal responses to hypoxia.

Previous reports from this laboratory have shown that a high percentage of neurons in the caudal hypothalamus are stimulated by hypoxia both in vivo and in vitro. This stimulation is in the form of an increase in firing frequency and significant membrane depolarization. The goal of the present study was to determine if this hypoxia-induced excitation is influenced by development. In addition, we sought to determine the mechanism by which hypoxia stimulates caudal hypothalamic neurons. Caudal hypothalamic neurons from neonatal (4-16 days) or juvenile (20-40 days) rats were patch-clamped, and the whole cell voltage and current responses to moderate (10% O2) or severe (0% O2) hypoxia were recorded in the brain slice preparation. Analysis of tissue oxygen levels demonstrated no significant difference in the levels of tissue oxygen in brain slices between the different age groups. A significantly larger input resistance, time constant and half-time to spike height was observed for neonatal neurons compared with juvenile neurons. Both moderate and severe hypoxia elicited a net inward current in a significantly larger percentage of caudal hypothalamic neurons from rats aged 20-40 days (juvenile) as compared with rats aged 4-16 days (neonatal). In contrast, there was no difference in the magnitude of the inward current response to moderate or severe hypoxia between the two age groups. Those cells that were stimulated by hypoxia demonstrated a significant decrease in input resistance during hypoxic stimulation that was not observed in those cells unaffected by hypoxia. A subset of neurons were tested independent of age for the ability to maintain the inward current response to hypoxia during synaptic blockade (11.4 mM Mg2+/0. 2 mM Ca2+). Most of the neurons tested (88.9%) maintained a hypoxic excitation during synaptic blockade, and this inward current response was unaffected by addition of 2 mM cobalt chloride to the bathing medium. In contrast, perfusion with the Na+ channel blocker, tetrodotoxin (1-2 microM) or Na+ replacement with N-methyl-D-glucamine (NMDG) significantly reduced the inward current response to hypoxia. Furthermore, the input resistance decrease observed during hypoxia was attenuated significantly during perfusion with NMDG. These results indicate the excitation elicited by hypoxia in hypothalamic neurons is age dependent. In addition, the inward current response of caudal hypothalamic neurons is not dependent on synaptic input but results from a sodium-dependent conductance.

Decrease in glutamic acid decarboxylase level in the hypothalamus of spontaneously hypertensive rats.

BACKGROUND: A reduction in gamma-aminobutyric (GABA)-mediated inhibition of pressor sites in the caudal hypothalamus of spontaneously hypertensive rats compared with that of normotensive Wistar-Kyoto rats has recently been demonstrated. OBJECTIVE: To determine whether the reduction in GABA-mediated inhibition of the caudal hypothalamus of the spontaneously hypertensive rats results from reductions both in the number of GABA-synthesizing neurons and in the amount of the GABA-synthesizing enzyme, glutamic acid decarboxylase messenger RNA (mRNA). DESIGN AND METHODS: A polyclonal antibody (Chemicon) for the 67 kDa isoform of glutamic acid decarboxylase (GAD67) was used to immunocytochemically label GABAergic neurons in the caudal hypothalamus of spontaneously hypertensive and Wistar-Kyoto rats that had been treated beforehand with colchicine. The labeled cells were counted for both strains by a blinded analysis and compared. Caudal hypothalamic tissues from spontaneously hypertensive and Wistar-Kyoto rats were analysed for GAD67 mRNA by Northern blotting. The signal intensities of the radioactive probe specific for GAD67 for the two strains were analyzed by using a phosphorimager and compared. Control areas for the immunocytochemical (zona incerta) and Northern blotting (cortex, midbrain, cerebellum, and brain stem) experiments were used to determine regional differences in expression of GAD67. RESULTS: Both the hypothalamus and cerebellum of spontaneously hypertensive and Wistar-Kyoto rats contained GAD67-immunoreactive neurons; however, there were 42% fewer GAD67 neurons in the caudal hypothalamus of spontaneously hypertensive rats than there were in that of Wistar-Kyoto rats. Furthermore, a 33% reduction in the amount of GAD67 messenger RNA in the caudal hypothalamus of spontaneously hypertensive rats compared with that for Wistar-Kyoto rats was demonstrated. Analysis of the expression of GAD67 in the cortex, midbrain, cerebellum, brain stem, and total brain revealed no difference between spontaneously hypertensive and Wistar-Kyoto rats. CONCLUSIONS: Our findings demonstrate that the spontaneously hypertensive rat has fewer neurons synthesizing GABA and less GAD67 mRNA in the caudal hypothalamus than do Wistar-Kyoto rats. This deficit in the GABAergic system in the caudal hypothalamus, a well-known cardiovascular regulatory site, could contribute to the essential hypertension in this animal model.

Neural control of the cardiovascular system during exercise. An integrative role for the vestibular system.

Precise regulatory signals are required in order to adjust the cardiovascular and respiratory systems to meet the demands of exercise. Two neural mechanisms, central command and a reflex originating in contracting muscles, are known to play a large role in exercise-associated adjustments in cardiovascular and respiratory activity. The extent to which other regulatory reflexes, such as vestibulo-autonomic reflexes, are able to impact upon the cardiovascular and respiratory systems during exercise is largely unknown. Further, brain regions that may integrate these control mechanisms are only starting to be investigated. We propose that medullary brain nuclei may integrate both exercise and vestibular signals to produce a more coordinated, and therefore efficient, means of adaptation to exercise in a gravitational environment.

Suprapontine control of respiration.

Despite focus on brainstem areas in central respiratory control, regions rostral to the medulla and pons are now recognized as being important in modulating respiratory outflow during various physiological states. The focus of this review is to highlight the role that suprapontine areas of the mammalian brain play in ventilatory control mechanisms. New imaging techniques have become invaluable in confirming and broadening our understanding of the manner in which the cerebral cortex of humans contributes to respiratory control during volitional breathing. In the diencephalon, the integration of respiratory output in relation to changes in homeostasis occurs in the caudal hypothalamic region of mammals. Most importantly, neurons in this region are strongly sensitive to perturbations in oxygen tension which modulates their level of excitation. In addition, the caudal hypothalamus is a major site for 'central command', or the parallel activation of locomotion and respiration. Furthermore, midbrain regions such as the periaqueductal gray and mesencephalic locomotor region function in similar fashion as the caudal hypothalamus with regard to locomotion and more especially the defense reaction. Together these suprapontine regions exert a strong modulation upon the basic respiratory drive generated in the brainstem.

Integrative role of medullary neurons of the cat during exercise.

The co-ordinated cardiovascular and respiratory responses observed during exercise are mediated by both afferent activation arising in contracting skeletal muscles and descending central drive originating in brain regions rostral to the medulla. Even though integration of these two mechanisms must occur in order to produce alterations in cardiorespiratory activity proportional to the intensity of the exercise, little is known about how this integration occurs. The purpose of the present study was to examine the role of medullary sites in the integration of exercise drives to the cardiovascular and respiratory systems. The responses of neurons in the ventrolateral medulla and the lateral tegmental fields to contraction of hindlimb muscles (elicited by stimulation of the L7 S1 ventral roots) and to activation of simulated central command (caudal hypothalamic stimulation) were examined in anaesthetized cats. Muscular contraction evoked an increase in discharge frequency in 59% (24/41) of the medullary neurons; 58% (14/24) of these neurons excited by muscular contraction displayed an immediate increase and 42% (10/24) had a more delayed increase in discharge frequency. Eighty-three per cent (10/12) of neurons rapidly excited by hindlimb muscle contraction were inhibited by hypothalamic stimulation. In contrast, most neurons which showed a delayed increase in discharge frequency during muscle contraction were either excited by hypothalamic stimulation (5/10) or unaffected (4/10). The majority of medullary neurons studied possessed a basal discharge related to the cardiovascular, respiratory and/or sympathetic cycles. These findings demonstrate that both feedback from contracting muscles and descending central command impinge upon individual medullary neurons. This is consistent with the idea that medullary neurons are important in integrating both the central drive and peripheral reflexes controlling the cardiorespiratory responses to exercise.

Oxygen-sensing neurons in the caudal hypothalamus and their role in cardiorespiratory control.

Work from this laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses to hypoxia. The purpose of this review is to describe the modulation of respiratory output by the caudal hypothalamus during hypoxia and how neurons in this area respond to hypoxia. The diaphragmatic activity response to hypoxia was significantly attenuated following microinjection of either cobalt chloride or kynurenic acid into the caudal hypothalamus of rats. In addition, caudal hypothalamic neurons in anesthetized rats and cats responded to hypoxia with an increased firing frequency. This response was maintained in the absence of input from the vagus and carotid sinus nerves in the cat. When recorded extracellularly or by whole-cell patch clamp in vitro, these neurons responded to hypoxia with an increase in firing frequency, membrane potential and inward current. These results suggest that the caudal hypothalamus exerts excitatory influence on respiration during hypoxia, that may originate from the ability of these neurons to sense changes in oxygen levels.

In vitro responses of VLM neurons to hypoxia after normobaric hypoxic acclimatization.

Hypoxic acclimatization involves an initial rapid ventilatory response followed by a more gradual increase in ventilation over a period of 24 to 48 h in both humans and rats. In addition, the acute ventilatory response to hypoxia is accentuated following hypoxic acclimatization. The purpose of the present investigation was to determine if hypoxic acclimatization augments the acute hypoxic response of neurons in the ventrolateral medulla (VLM). Brain slices (400 microns) containing the ventrolateral medulla were prepared from Sprague-Dawley rats acclimatized to hypoxia (10% O2) for 4-5 days (n = 4) and 9-10 days (n = 4) and from rats maintained in a normoxic environment (n = 4). Extracellular recordings demonstrated that there were no significant differences in the basal pattern or discharge rate of VLM neurons from animals exposed to short (10.8 +/- 0.9 Hz, n = 51), or long (10.1 +/- 1.1 Hz, n = 59) periods of hypoxia compared to control neurons (10.8 +/- 1.1 Hz, n = 52). The proportion of neurons stimulated (approximately 70%), inhibited (approximately 20%) and unaffected (approximately 10%) by an acute bout of hypoxia (10% O2) was also similar among groups. However, acute hypoxia elicited a greater increase in discharge frequency in neurons from rats exposed to the short period of hypoxia compared to the responses from neurons in the control and longer acclimatization groups. Thus, the responsivity of VLM neurons during the early stages of hypoxic acclimatization is altered in a manner consistent with the respiratory responses associated with acclimatization.

Identification of diencephalic and brainstem cardiorespiratory areas activated during exercise.

The purpose of this study was to identify diencephalic and brainstem sites active during exercise (EX) in conscious rats running on a treadmill. Brain areas active during exercise, compared to rest conditions (non-EX), were identified using immunocytochemical labelling of the protein product of the proto-oncogene c-fos. Increased labelling was observed in the 'defence area' or 'hypothalamic/subthalamic locomotor regions' including the posterior and lateral hypothalamic areas. Increased labelling with EX was found in both colliculi, the periaqueductal gray matter, the parabrachial complex and the cuneiform nucleus ('mesencephalic locomotor region'). Increased labelling with EX was also found in the medial portion of n. tractus solitarius, and both the rostral and caudal ventrolateral medulla. Conspicuous by an absence of labelling during EX were cells in thalamic areas associated with somatosensory function, although the dorsal column nuclei were also labelled above control. Thus, areas in which labelling was increased during exercise closely correlate with the brain areas which have been implicated in both autonomic and somatomotor control. These results from awake, exercising rats support those obtained previously in anesthetized animal preparations.

Ventrolateral medullary neurons show age-dependent depolarizations to hypoxia in vitro.

Central mechanisms are likely responsible for the larger respiratory activation in response to hypoxia in the adult compared to the neonatal animal. One possible site for this effect is in the ventrolateral medulla, an area known to be involved in the cardiorespiratory responses to hypoxia. Neurons in this area are stimulated by hypoxia both in vivo and in vitro. The purpose of the present study was to determine if changes in the magnitude of this excitatory response occur during early postnatal development. Whole-cell patch recordings were made from neurons in the ventrolateral medulla (VLM) in a 400-micrograms brain slice preparation. The basal properties and responses to a brief (90 s) hypoxic stimulus (5% CO2/95% N2) were compared between neurons from neonatal (P < 16) and juvenile (P16-38) rats. An excitation consisting of a depolarization, increase in spike frequency and decrease in input resistance was observed during hypoxia in eighty-three percent of juvenile but in only 58% of the neonatal VLM neurons. Moreover, the magnitude of this response was greater in the juvenile (8.2 +/- 1.3 mV) than in the neonatal (4.8 +/- 0.5 mV) neurons. A second type of depolarizing response, consisting of a more pronounced depolarization interrupted by a brief hyperpolarization that returned to a depolarized state and not associated with an increased discharge frequency, occurred in only 3% of the neurons from the juvenile animals compared to 18% of those from neonatal rats. The remaining proportion of the VLM neurons studied were hyperpolarized or were unaffected by hypoxia. Measurements of tissue pO2 indicate that none of the above differences are due to variations in the hypoxic stimulus between neonatal and adult slices. The results of this study suggest that the hypoxic-induced depolarizations observed in VLM neurons change during development. These developmental changes may contribute to the changes that occur in cardiorespiratory responses to acute systemic hypoxia during early development.