0066 Glycinergic Postsynaptic Inhibition is Responsible for the Suppression of Hypoglossal Motoneuron Activity During Naturally-Occurring REM Sleep

Introduction The present study was undertaken to explore the role of glycinergic postsynaptic inhibition and monoaminergic disfacilitation (a withdrawal of excitatory noradrenergic and serotonergic inputs) in the control of hypoglossal motoneuron activity during REM sleep. Accordingly, glycinergic, noradrenergic and serotonergic antagonists were microinjected into the hypoglossal nucleus, and their effects on the hypoglossal nerve activity during REM sleep were examined in chronically-instrumented, unanesthetized cats. Methods Adults cats were prepared for monitoring behavioral states of sleep and wakefulness, and for extracellular recordings from hypoglossal nerve. Strychnine (a glycinergic antagonist) and a mixture of prazosin (a noradrenergic antagonist) and methysergide (a serotonergic antagonist) were microinjected, separately, into the hypoglossal nucleus during naturally-occurring states of sleep and wakefulness. Results During REM sleep, compared to non-REM sleep, the hypoglossal nerve activity decreased by 17.4±1.5% (n=17) in the control recordings (prior to the injection of strychnine). Following the microinjection of strychnine, there was only a mean decrease of 7.2±1.2% (n=12) in the nerve activity during REM sleep versus NREM sleep. The strychnine effect was statistically significant compared to control (p<0.001; unpaired t-test), which indicates that strychnine blocks REM sleep-related suppression of hypoglossal nerve activity. In contrast, the microinjection of prazosin and methysergide did not significantly reduce the hypoglossal nerve activity during REM sleep (control: 15.9±2.3, n=9 vs. prazosin+methysergide: 12.6±1.4%, n=10, p=0.229, unpaired t-test). Conclusion The present results demonstrate that the microapplication of strychnine, but not prazosin and methysergide, into the hypoglossal nucleus significantly reduces the suppression of the hypoglossal nerve activity during naturally-occurring REM sleep. We therefore suggest that glycinergic postsynaptic inhibition is primarily responsible for the suppression of hypoglossal motoneuron activity during REM sleep. Support 5R01NS094062

Differential activating effects of thyrotropin-releasing hormone and its analog taltirelin on motor output to the tongue musculature in vivo

Thyrotropin-releasing hormone (TRH) is produced by the hypothalamus but most brain TRH is located elsewhere where it acts as a neuromodulator. TRH-positive neurons project to the hypoglossal motoneuron pool where TRH receptor RNA shows a high degree of differential expression compared with the rest of the brain. Strategies to modulate hypoglossal motor activity are of physiological and clinical interest given the potential for pharmacotherapy for obstructive sleep apnea (OSA), a common and serious respiratory disorder. Here, we identified the effects on tongue motor activity of TRH and a specific analog (taltirelin) applied locally to the hypoglossal motoneuron pool and systemically in vivo. Studies were performed under isoflurane anesthesia and across sleep–wake states in rats. In anesthetized rats, microperfusion of TRH (n = 8) or taltirelin (n = 9) into the hypoglossal motoneuron pool caused dose-dependent increases in tonic and phasic tongue motor activity (both p < 0.001). However, the motor responses to TRH were biphasic, being significantly larger “early” in the response versus at the end of the intervention (p ≤ 0.022). In contrast, responses to taltirelin were similar “early” versus “late” (p ≥ 0.107); i.e. once elicited, the motor responses to taltirelin were sustained and maintained. In freely behaving conscious rats (n = 10), microperfusion of 10 μM taltirelin into the hypoglossal motoneuron pool increased tonic and phasic tongue motor activity in non-rapid-eye-movement (REM) sleep (p ≤ 0.038). Intraperitoneal injection of taltirelin (1 mg/kg, n = 16 rats) also increased tonic tongue motor activity across sleep–wake states (p = 0.010). These findings inform the studies in humans to identify the potential beneficial effects of taltirelin for breathing during sleep and OSA.

Synchronization of presynaptic input to motor units of tongue, inspiratory intercostal, and diaphragm muscles

The respiratory central pattern generator distributes rhythmic excitatory input to phrenic, intercostal, and hypoglossal premotor neurons. The degree to which this input shapes motor neuron activity can vary across respiratory muscles and motor neuron pools. We evaluated the extent to which respiratory drive synchronizes the activation of motor unit pairs in tongue (genioglossus, hyoglossus) and chest-wall (diaphragm, external intercostals) muscles using coherence analysis. This is a frequency domain technique, which characterizes the frequency and relative strength of neural inputs that are common to each of the recorded motor units. We also examined coherence across the two tongue muscles, as our previous work shows that, despite being antagonists, they are strongly coactivated during the inspiratory phase, suggesting that excitatory input from the premotor neurons is distributed broadly throughout the hypoglossal motoneuron pool. All motor unit pairs showed highly correlated activity in the low-frequency range (1–8 Hz), reflecting the fundamental respiratory frequency and its harmonics. Coherence of motor unit pairs recorded either within or across the tongue muscles was similar, consistent with broadly distributed premotor input to the hypoglossal motoneuron pool. Interestingly, motor units from diaphragm and external intercostal muscles showed significantly higher coherence across the 10–20-Hz bandwidth than tongue-muscle units. We propose that the lower coherence in tongue-muscle motor units over this range reflects a larger constellation of presynaptic inputs, which collectively lead to a reduction in the coherence between hypoglossal motoneurons in this frequency band. This, in turn, may reflect the relative simplicity of the respiratory drive to the diaphragm and intercostal muscles, compared with the greater diversity of functions fulfilled by muscles of the tongue.