Karen B. Bach

Hypoxia-induced long-term facilitation of respiratory activity is serotonin dependent

Repeated isocapnic hypoxia evokes long-term facilitation (LTF) of phrenic nerve activity in rats. We wished to determine: (1) whether hypoxia-induced LTF is serotonin dependent; and (2) whether hypoxia-induced LTF is a property of upper airway motoneurons. Phrenic and hypoglossal nerve activities were recorded in urethane anesthetized, vagotomized, paralyzed and artificially ventilated rats (n = 7). Rats were exposed to three, 5-min hypoxic episodes (FIo2 = 0.10) separated by 5 min of hyperoxia (FIo2 = 0.50). One hour after the final hypoxic episode, integrated phrenic and hypoglossal amplitudes and burst frequency were increased above control values (63 +/- 17%, 78 +/- 26% and 9.6 +/- 2.1 bursts/min, respectively: p < 0.05). In rats pretreated with methysergide (n = 7; 4 mg/kg), no changes in phrenic or hypoglossal activity from pre-stimulus control values were observed at any time post-stimulation. The results indicate that hypoxia-induced LTF requires 5-HT receptors and is characteristic of both hypoglossal and phrenic motor output.

Long term facilitation of phrenic motor output

Episodic hypoxia or electrical stimulation of carotid chemoafferent neurons elicits a sustained, serotonin-dependent augmentation of respiratory motor output known as long term facilitation (LTF). The primary objectives of this paper are to provide an updated review of the literature pertaining to LTF, to investigate the influence of selected variables on LTF via meta-analysis of a large data set from LTF experiments on anesthetized rats, and to propose an updated mechanism of LTF. LTF has been demonstrated in anesthetized and awake experimental preparations, and can be evoked in some human subjects during sleep. The mechanism underlying LTF requires episodic chemoafferent stimulation, and is not elicited by similar cumulative durations of sustained hypoxia. Meta-analysis of phrenic nerve responses following episodic hypoxia in 63 experiments on anesthetized rats (conducted by four investigators over a period of several years) indicates that phrenic LTF magnitude correlates with peak phrenic responses during hypoxia and hypercapnia, but not with the level of hypoxia during episodic exposures. Potential mechanisms underlying these relationships are discussed, and currently available data are synthesized into an updated mechanistic model of LTF. In this model, we propose that LTF arises predominantly from episodic activation of serotonergic receptors on phrenic motoneurons, activating intracellular kinases and, thus, phosphorylating and potentiating ionic currents associated with the glutamate receptors that mediate respiratory drive.

Post-hypoxia frequency decline in rats: sensitivity to repeated hypoxia and α2-adrenoreceptor antagonism

We tested the hypothesis that the post-hypoxia frequency decline of phrenic nerve activity following brief, isocapnic hypoxic episodes in rats is diminished by prior hypoxic episodes and alpha2-adrenoreceptor antagonism. Anesthetized (urethane), artificially ventilated (FIO2=0.50) and vagotomized rats were presented with two or three, 5 min episodes of isocapnic hypoxia (FIO2 approximately 0.11), separated by 30 min of control, hyperoxic conditions. Phrenic nerve discharge, end-tidal CO2, and arterial blood gases were measured before during and after hypoxia. The average maximum frequency decline, measured 5 min after the first hypoxic episode, was 26+/-7 bursts/min below pre-hypoxic baseline values (a 70+/-16% decrease). By 30 min post-hypoxia, frequency had returned to baseline. Two groups of rats were then administered either: (1) saline (sham) or (2) the alpha2-receptor antagonist, RX821002 HCl (2-[2-(2-Methoxy-1,4-benzodioxanyl)] imidazoline hydrochloride; 0.25 mg/kg, i.v.). Isocapnic hypoxia was repeated 10 min later. In sham rats, the post-hypoxia frequency decline (PHFD) was significantly attenuated relative to the initial (control) response. However, PHFD was attenuated significantly more in RX821002-treated vs. sham rats (-3+/-3 bursts/min vs. -12+/-4 bursts/min @ 5 min post hypoxia for RX821002 and sham-treated, respectively; p<0.05). We conclude that the magnitude of PHFD is dependent on the prior history of hypoxia and that alpha2 adrenoreceptor activation plays a role in its underlying mechanism.

Serotonin reveals ineffective spinal pathways to contralateral phrenic motoneurons in spinally hemisected rats

Serotonin reveals ineffective (subthreshold) pathways from the C2 lateral funiculus to ipsilateral phrenic motoneurons in spinalized rats. The objective of the present study was to investigate serotonergic modulation of crossed-spinal pathways to contralateral phrenic motoneurons. Rats (n = 10) were anesthetized (urethane), paralyzed, vagotomized, and artificially ventilated. The spinal cord was hemisected at C1–C2 and, on the intact side, a tungsten stimulating electrode was placed ventral to the C2 dorsal root entry zone in the dorsolateral (∼ 1.1 mm) or the ventrolateral funiculus (∼2.2 mm depth). Single shocks (100–750 μA, 0.1–0.5 ms, 2 Hz) elicited a short-latency (∼ 1.0 ms to peak) excitation in the ipsilateral phrenic nerve, but usually evoked little or no response in the contralateral phrenic nerve at either stimulus site. Following systemic injection of the monoamine oxidase inhibitor pargyline (25 mg/kg) and the serotonin precursor 5-hydroxytryptophan (5–10 mg/kg), complex responses were revealed in the contralateral phrenic nerve, including; (1) spontaneous tonic activity; (2) a short-latency (∼1.0 ms to peak) evoked excitation; and (3) two long-latency (∼2.2 and 7.8 ms to peak) evoked excitations. The longest latency excitation was expressed only when the stimulating electrode was positioned in the dorsolateral funiculus. Contralateral evoked responses were blocked by systemic methysergide (2–6 mg/kg), a broad-spectrum serotonin receptor antagonist. These results indicate that serotonin converts ineffective crossed phrenic pathways in the spinal cord to effective pathways. It remains to be determined whether serotonin is both necessary and sufficient in this modulatory process, or if it is a nonspecific result of increased phrenic motoneuron excitability.

Modulation of ventilatory control during exercise

The control of ventilatory responses to mild or moderate dynamic exercise has been the subject of considerable debate for over a century. The prevailing view has been that the ventilatory response to exercise is stereotypical and rather unmalleable. However, paradigms involving novel associations of stimulus inputs have been shown to modulate breathing in short and longer time scales. The scope of this review includes examples of modified ventilatory responses to exercise which have been investigated in terms of neural mechanisms. An attempt to synthesise the available data into a model of neuromodulation is presented.

Serotonin is necessary for short-term modulation of the exercise ventilatory response

The exercise ventilatory response is augmented during conditions of increased respiratory dead space (delta Vd), a phenomenon that we refer to as short term modulation (STM). To test the hypothesis that serotonin is necessary in the mechanism underlying STM, experiments were conducted on ten awake goats. Ventilation, CO2 production and PaCO2 were measured at rest and during treadmill exercise (4 km/h, 5% grade), with and without delta Vd (0.25 L), before and after systemic administration of the serotonin receptor antagonist, methysergide maleate (n = 6; 1 mg/kg, i.v.), or the tryptophan hydroxylase inhibitor, p-chlorophenylalanine (PCPA; n = 4; 100 mg/kg, i.v.). Pre-methysergide: (1) PaCO2 decreased from rest to exercise to a similar degree with (-1.9 mmHg) and without (-1.8 mmHg) delta Vd; (2) the exercise ventilatory response increased 59% +/- 13% (P < 0.01) with delta Vd, accounting for similar exercise PaCO2 regulation and demonstrating STM; and (3) effects of delta Vd on exercise tidal volume and frequency responses were inconsistent. Post-methysergide: (1) there were no significant effects on ventilation or PaCO2 at rest or during exercise in control (mask) conditions; (2) the exercise ventilatory response was unaffected by delta Vd, thereby allowing PaCO2 to increase 4.1 +/- 3.0 mmHg from rest to exercise (P < 0.05); and (3) with delta Vd during exercise, the tidal volume response was increased, but was offset by a decreased frequency response. Following PCPA (16-24 h): (1) hyperventilation was evident at rest and during exercise; (2) the exercise ventilatory response was augmented, indicating STM; and (3) the exercise ventilatory response with delta Vd was not affected further, allowing PaCO2 to increase from rest to exercise and indicating an inability to elicit further STM. These data suggest that serotonin is necessary for short term modulation of the exercise ventilatory response with increased respiratory dead space, although the location of relevant serotonin receptors is not yet clear.

Phrenic responses to contralateral spinal stimulation in rats: effects of old age or chronic spinal hemisection.

Serotonin reveals ineffective spinal pathways from the C2-lateral funiculus to contralateral phrenic motoneurons in young adult rats with acute spinal hemisection. We tested the hypothesis that old age (1.5-2 years) or chronic hemisection (3-5 days) strengthens these pre-existing crossed spinal pathways. There were no consistent differences between young adult rats with acute hemisection versus young adult rats with chronic hemisection or old rat with acute hemisection except that one long-latency phrenic excitation could not be elicited in old rats. The results indicate that neither old age nor chronic hemisection strengthens crossed-spinal pathways, but that old age may selectively diminish spinal pathways involved in the neural control of breathing.