Kurt Schmid

Inhibitions mediated by glycine and GABAA receptors shape the discharge pattern of bulbar respiratory neurons

Experiments were performed to identify the glycinergic or GABAergic nature, and the timing of discharge, of the neurons which produce chloride-dependent inhibitions on other bulbar respiratory neurons (RNs) during their silent and active phases. RNs recorded extracellularly in pentobarbital-anesthetized or decerebrate cats, were subjected to iontophoretic applications of glutamate, of the glycine antagonist strychnine, and of the GABAA receptor antagonist bicuculline. Both antagonists induced discharge or increased discharge frequency in restricted parts of the respiratory cycle without affecting the discharge frequency in other parts of the cycle. Strychnine most often elicited activity in late-inspiration and early-expiration, but also in early inspiration and in late expiration. Bicuculline was most often effective throughout the entire discharge period of each neuron with no effect during the silent period, although it also acted selectively during late-inspiration in inspiratory neurons, an effect attributed to GABAA receptor blockade. The convergence of glycinergic afferent inputs during late inspiration and early expiration suggests that glycinergic neurons may play an important role in the inspiratory to expiratory phase transition.

Glycine receptor-mediated fast synaptic inhibition in the brainstem respiratory system

The involvement of glycinergic neurotransmission in the central regulation of respiration was investigated by administration of specific receptor agonists and antagonists into the 4th cerebral ventricle of the rabbit. Central respiratory activity was assessed by cycle-triggered averaging of phrenic nerve activity (PNA). Glycine reduced the peak amplitude of PNA and increased the duration of the respiratory phases. Conversely, strychnine decreased the duration of both phases, being more effective in expiration. Strychnine converted inspiratory ramp activity to a decrescendo type of pattern, with the highest discharge activity present at the onset of the inspiratory phase. Strychnine induced tonic PNA lasting throughout expiration with a decrescendo type of activity. In many cases strychnine induce apneusis with tonic PNA being at a level of about 2/3 of its maximum value during control. We conclude that glycinergic neurotransmission is important for the maintenance of the respiratory rhythm as well as the generation of normal respiratory pattern. Glycine could be involved in phasic as well as tonic inhibitory neurotransmission within the central respiratory system.

Serotonergic control of phrenic motoneuronal activity at the level of the spinal cord of the rabbit

The role of serotonin (5-hydroxytryptamine, 5-HT)-mediated modulation of phrenic motoneuronal activity was evaluated by microapplication of 5-HT and methysergide into the phrenic nuclei of the rabbit. 5-HT facilitated phrenic nerve activity (PNA) considerably resulting in a long-lasting augmentation of the peak amplitude of integrated PNA. In contrast, the blockade of intrinsically active 5-HT by methysergide decreased PNA and led to a strong reduction of the peak amplitude of integrated PNA. MDL 72222 was ineffective. Blockade of 5-HT receptors by preceding administration of methysergide effectively abolished the effects of microinjected 5-HT. Respiratory timing was unaffected by both the agonist and the antagonist. These results suggest that a considerable portion of the facilitatory influence of caudal raphe nuclei on central respiratory activity takes place at the phrenic nuclei level.

Evidence for an involvement of NMDA and non-NMDA receptors in synaptic excitation of phrenic motoneurons in the rabbit.

The action of endogenous excitatory amino acids on phrenic motoneurons was studied in anesthetized, vagotomized, paralyzed and artificially ventilated rabbits. The NMDA receptor antagonists APV and ketamine, as well as the non-NMDA receptor antagonists GAMS and DNQX were administered by microinjection into the ventral horn of the spinal segments C3-C5. Injection of each antagonist resulted in a reversible reduction of the phrenic nerve activity. Results suggest an important function of endogenous excitatory amino acids in the excitation of phrenic motoneurons. NMDA as well as non-NMDA receptors are involved. The functional role of both receptor types in bulbospinal neurotransmission is discussed.

Medullary respiratory-related neurons with axonal connections to rostral pons and their function in termination of inspiration

In urethane-anaesthetized, paralyzed and artificially ventilated rabbits, medullary respiration-related neurons (RRU) were classified according to the phase relation of their burst discharge to phrenic nerve activity. Phase-bound inspiratory (I) or expiratory (E) neurons were discriminated from phase-spanning expiratory-inspiratory (EI) or inspiratory-expiratory (IE) units. Mechanisms of termination of inspiration by electrical stimulation of rostral pontine nuclei (Nc. parabrachialis medialis; Lc. coeruleus) were examined firstly to demonstrate whether RRU receive descending excitatory and inhibitory afferents as well as ascending efferents and secondly to analyse the time course of the neuronal pathways involved. Of 120 RRU, 38 neurons were demonstrated to receive pontine afferents. About 33% of all E neurons became orthodromically excited during rostral pons stimulation whereas 18.2% of all I cells became orthodromically inhibited. Some RRU were shown to project up to the rostral pons. 50% of these were of the phase-spanning IE type. The onset of inspiratory inhibition induced by rostral pons stimulation occurred 3.4 ms after the onset of single electrical pulse stimulation. Based on these results a neuronal model for a pontine mechanism terminating inspiration is proposed.

GABAB receptor mediated effects on central respiratory system and their antagonism by phaclofen

The role of GABAB receptors in control of central respiratory system was evaluated by cycle-triggered averaging of phrenic nerve activity (PNA) of the rabbit. Blockade of GABAB receptors of the caudal brainstem by intracerebroventricular administration of phaclofen augmented PNA, decreased the duration of inspiration and to about the same extent increased the duration of expiration thus unmasking intrinsically active GABA. Analogously, stimulation of brainstem GABAB receptors by exogenous baclofen decreased PNA. Preceding administration of larger doses of phaclofen could block the effects of baclofen. It is proposed that GABAB receptors are involved in tonic and phasic modulation of central respiratory activity.

Effects of corticotropin-releasing factor on central respiratory activity.

Effects of corticotropin-releasing factor (CRF) on central regulation of respiration were studied in urethane-anaesthetized, vagotomized, paralyzed and artificially ventilated rabbits. Injections of CRF into the IVth cerebral ventrile (i.c.v.) resulted in an increase of the neuronal tidal volume (nVt), but had only minor effects on the duration of respiratory phases. Microinjection of CRF into the ventral parabrachial region of the pons (pneumotaxic center) resulted in a reduction of nVt and of the respiration rate. Respiratory effects of CRF administered i.c.v. or into the pontine parabrachial region were antagonized by alpha-helical-CRF, indicating a receptor-mediated action of CRF. Respiratory effects of CRF were not significantly affected after blocking ganglionic transmission with chlorisondamine. Neither mean arterial blood pressure nor heart rate were significantly affected by the injection of CRF into the IVth ventricle or the rostral pons. Results suggest an involvement of CRF in the central regulation of respiratory movements. CRF may be involved in the adaptation of respiration to stress.

Evidence for a respiration-modulated cholinergic action on the activity of medullary respiration-related neurons in the rabbit

Effects of the iontophoretically administered cholinergic agonists acetylcholine, bethanechol and DMPP on the activity of medullary respiration-related neurons were examined in urethane-anaesthetized rabbits. Inhibitory effects prevailed over excitatory effects. Analysis of cholinergic effects by cycle-triggered averaging revealed three major types of neuronal responses: (i) constant alterations of spike-density throughout the whole period of activity (“constant effects”), (ii) effects increasing during the progression of the burst of discharge or effects restricted to a particular fraction of the burst (“phasic effects”) and (iii) effects which were characterized by an excitation during one respiratory phase and an inhibition during the other phase (“bi-phasic effects”). The latter type of effects was observed in phase-spanning respiration-related neurons. Phasic effects were mainly observed in inspiration-related neurons which were predominantly inhibited by stimulation of muscarinic receptors. Inspiratory Rβ-neurons in no case were phasically affected by cholinergic agents. The mean muscarinic inhibition of inspiration-related neurons increased with the progression of inspiration. The mean nicotinic inhibition of expiration-related neurons decreased with the progression of expiration. Results suggest that the efficacy of (i) a central inspiration terminating mechanism and (ii) the onset of discharge of expiratory neurons is modulated by acetylcholine.

Involvement of fast synaptic inhibition in the generation of high-frequency oscillation in central respiratory system.

During moderate hypercapnia, spectrum analysis of efferent phrenic nerve activity (PNA) of urethane anesthetized rabbits revealed high-frequency oscillations (HFO). The spectral peak (mean frequency 111 Hz) was reversibly eliminated by the glycine receptor antagonist strychnine administered into the 4th cerebral ventricle. In contrast, blockade of brainstem GABAA receptors by bicuculline did not abolish HFO but even reinforced or induced HFO. Thus, there is evidence that fast synaptic inhibition via glycine receptors is involved in the generation of HFO within the medullary respiratory center, whereas GABA may play a modulatory role.

Cycle-triggered averaging of respiration-related neuronal activity.

A computer system is presented which provides off-line computation of cycle-triggered histograms (CTH) of respiration-related neuronal activity. Binwidths of the histograms are freely selectable by software from 10 ms to 100 ms. For special evaluation purposes, CTHs can be standardized in different ways concerning cycle duration as well as amplitude. Time incidence of maximum frequency, center of gravity and expiration-to-inspiration phase transition within the respiratory cycle are computed. The system employs special hardware interfaces to an 8-bit microcomputer which are briefly described. Data acquisition, data manipulation and output handling of the results are performed by chaining 3 compiled BASIC programs. Some comments on peculiarities of the BASIC language concerning combined application of a BASIC interpreter and a BASIC compiler are brought up. The usefulness of the method is demonstrated by examples of CTHs computed from the activity of medullary respiration-related neurons as well as of the corresponding phrenic nerve mass activity.