C. Larraín

Effects of dopaminergic blockade upon carotid chemosensory activity and its hypoxia-induced excitation

The effects of domperidone, antagonist of D2 receptors, on arterial chemoreceptor activity were studied in spontaneously breathing and pentobarbitone anesthetized cats, in which recordings of chemosensory impulse activity were obtained simultaneously from both cut carotid (sinus) nerves. Intravenous injections of domperidone 50 micrograms/kg produced a maintained increase in the basal frequency of chemosensory discharges, after which hyperoxic tests (breathing 100% O2 for 30 s) evoked larger falls in the rate of chemosensory impulses. Chemosensory responses evoked by hypoxic hypoxia (100% N2 tests) and by cytotoxic hypoxia (i.v. injections of NaCN) reached higher impulse rates after domperidone treatment. The effects of domperidone reveal that a resting release of dopamine from glomus cells maintains a low level of basal chemosensory activity under normoxic conditions. Domperidone turns off such restraining dopaminergic control and enhances the transient chemosensory responses to hypoxic stimuli. Present data support a modulatory role for dopamine within the chemoreceptor process, but not its participation as excitatory transmitter between glomus cells and sensory nerve endings.

Lipopolysaccharide‐induced carotid body inflammation in cats: functional manifestations, histopathology and involvement of tumour necrosis factor‐α

In the absence of information on functional manifestations of carotid body (CB) inflammation, we studied an experimental model in which lipopolysaccharide (LPS) administration to pentobarbitone‐anaesthetized cats was performed by topical application upon the CB surface or by intravenous infusion (endotoxaemia). The latter caused: (i) disorganization of CB glomoids, increased connective tissue, and rapid recruitment of polymorphonuclear cells into the vascular bed and parenchyma within 4 h; (ii) increased respiratory frequency and diminished ventilatory chemoreflex responses to brief hypoxia (breathing 100% N2 for 10 s) and diminished ventilatory chemosensory drive (assessed by 100% O2 tests) during normoxia and hypoxia; (iii) tachycardia, increased haematocrit and systemic hypotension in response to LPS i.v.; and (iv) increased basal frequency of carotid chemosensory discharges during normoxia, but no change in maximal chemoreceptor responses to brief hypoxic exposures. Lipopolysaccharide‐induced tachypnoea was prevented by prior bilateral carotid neurotomy. Apoptosis was not observed in CBs from cats subjected to endotoxaemia. Searching for pro‐inflammatory mediators, tumour necrosis factor‐α (TNF‐α) was localized by immunohistochemistry in glomus and endothelial cells; reverse transcriptase‐polymerase chain reaction revealed that the CB expresses the mRNAs for both type‐1 (TNF‐R1) and type‐2 TNF‐α receptors (TNF‐R2); Western blot confirmed a band of the size expected for TNF‐R1; and histochemistry showed the presence of TNF‐R1 in glomus cells and of TNF‐R2 in endothelial cells. Experiments in vitro showed that the frequency of carotid nerve discharges recorded from CBs perfused and superfused under normoxic conditions was not significantly modified by TNF‐α, but that the enhanced frequency of chemosensory discharges recorded along responses to hypoxic stimulation was transiently diminished in a dose‐dependent manner by TNF‐α injections. The results suggest that the CB may operate as a sensor for immune signals, that the CB exhibits histological features of acute inflammation induced by LPS, that TNF‐α may participate in LPS‐induced changes in chemosensory activity and that some pathophysiological reactions to high levels of LPS in the bloodstream may originate from changes in CB function.

Effects of combined cholinergic–purinergic block upon cat carotid body chemoreceptors in vitro

Since acetylcholine (ACh) and ATP have been proposed as excitatory co-transmitters at synapses between glomus cells and sensory nerve endings of the carotid body (CB), we tested such hypothesis by studying the effects of combined cholinergic-purinergic block on the chemosensory activity recorded from cats carotid bodies perfused and/or superfused in vitro. The preparations were bathed with Tyrodes solution, either normoxic (PO2=98.5+/-13.5 Torr) or hypoxic (PO2=31.8+/-5.2 Torr), and the frequency of chemosensory impulses (fchi) was recorded from the carotid (sinus) nerve. Dose-response curves for fchi increases evoked by intra-stream boluses of acetylcholine, nicotine and ATP were studied. A combination of mecamylamine 2 microM and suramin 50 microM, applied through the perfusate or superfusate, suppressed nicotine- and ATP-induced increases in fchi, but the basal chemosensory activity in normoxia and the chemosensory excitation elicited by hypoxic superfusion were preserved, although variably reduced in most preparations. Thus, in spite of the excitatory effects provoked by applying ACh and ATP to the perfused/superfused CB in vitro, a co-release of these substances cannot account entirely for the chemosensory excitation induced by hypoxic stimulation of the CB.

Immunosensory signalling by carotid body chemoreceptors

Injections of lipopolysaccharide (LPS) have been used to produce the signs of sepsis and study their underlying mechanisms. Intravenous (IV) injections of LPS in anesthetized cats induce tachypnea, tachycardia and hypotension, but ventilatory changes are suppressed after sectioning carotid and aortic nerves. Otherwise, LPS increases the basal frequency of carotid chemosensory discharges, but reduces ventilatory and chemosensory responses to hypoxia and nicotine injections. Increases in cytokines (IL-1β, IL-6 and TNF-α) are observed in plasma and tissues after injecting LPS. In carotid bodies perfused in vitro, TNF-α reduces chemosensory discharges induced by hypoxia. The rat carotid body and its sensory ganglion constitutively express LPS canonical receptor, TLR4, as well as TNF-α and its receptors (TNF-R1 and TNF-R2). Increases of TNF-α and TNF-R2 expression occur after LPS administration. The activation of peripheral and central autonomic pathways induced by LPS or ILs is partly dependent on intact vagus nerves. Thus, the carotid and vagus nerves provide routes between the immune system and CNS structures involved in systemic inflammatory responses.

Carotid body chemosensory activity and ventilatory chemoreflexes in cats persist after combined cholinergic-purinergic block.

Acetylcholine (ACh) and ATP have been proposed as excitatory co-transmitters operating at synapses between glomus cells and sensory nerve endings of the carotid body (CB). To test such hypothesis, we performed experiments on cats under pentobarbitone anesthesia and breathing spontaneously. Cholinergic and purinergic agonists and antagonists were given into one common carotid artery. Chemoreflex ventilatory changes initiated from the ipsilateral CB or chemosensory activity from the ipsilateral carotid nerve were recorded. Agonists ACh, nicotine, epibatidine, ATP, betagamma-methylene-ATP and gammaS-ATP induced transient chemoreflex enhancements of ventilation or increased chemosensory activity. When given in combination, mecamylamine and suramin suppressed both nicotine- and ATP-induced ventilatory chemoreflexes or chemosensory responses. However, neither chemoreflex hyperventilation induced by brief hypoxic exposures or steady-state hypoxic levels, nor chemosensory excitation elicited by these maneuvers were eliminated. Asphyxia-induced chemosensory excitation was not reduced by combined blockade of ACh and ATP receptors. Furthermore, ventilatory or chemosensory depression evoked by 100% O2 tests was unmodified, thus evidencing that basal chemosensory drive in normoxia was not suppressed by combined cholinergic-purinergic blockade. Therefore, although ACh and ATP may participate in chemoexcitation of the CB, their involvement fails to explain the origin of chemosensory discharges from synaptic transmission between glomus cells and chemosensory nerve endings of the CB.

Acute ventilatory and circulatory reactions evoked by nicotine: are they excitatory or depressant?

Either excitatory or inhibitory cardio-respiratory responses induced by nicotine have been reported. We evaluated the joint and separate contributions of peripheral arterial chemoreceptors and pulmonary vagal afferences to nicotine-induced cardio-respiratory responses in 11 pentobarbitone-anaesthetized cats. Nicotine, given i.v. in doses of from 1 to 200 microg/kg, evoked dose-dependent transient increases in tidal volume (VT) and arterial blood pressure (BP), but the highest doses evoked brief apnoea, immediately followed by intense hyperventilation, as well as discrete early hypotension followed by late hypertension. Bilateral section of the aortic and carotid nerves abolished all hyperventilatory responses to nicotine, giving way to apnoea followed by few cycles of reduced VT and profound hypotension followed by slight hypertension in response to intermediate doses (50-100 microg/kg). Subsequent bilateral vagotomy (BV) suppressed apnoeic and hypotensive responses. In other cats initially subjected to BV, only increases in VT and BP were observed in response to nicotine, effects which were no longer observed after additional carotid and aortic deafferentation. These data suggest that excitatory effects of nicotine on respiration and BP are reflexes evoked by stimulation of peripheral arterial chemoreceptors, while inhibitory effects are also reflex responses but evoked from stimulation of pulmonary vagal afferences.

Thermal effects on ventilation in cats: participation of carotid body chemoreceptors

In pentobarbitone anesthetized cats, raising body temperature from 37 to 40 degrees C by external heat increased respiratory frequency, tidal volume, frequency of spontaneous gasps and mean inspiratory flow. It reduced end-tidal CO2 pressure, together with inspiratory and expiratory durations. After bilateral section of the carotid nerves, raising body temperature still induced hyperventilation, but the increase in gasp frequency was less pronounced and no significant change in tidal volume was observed. In comparison to steady ventilatory values in the intact condition, significant reductions in tidal volume at 38 degrees C and in gasp frequency at 37, 39 and 40 degrees C were observed after bilateral carotid neurotomy. Brief hyperoxic tests induced transient decreases in tidal volume and increases in end-tidal CO2 pressure which were significantly larger at 40 degrees C than at 37 degrees C. These changes disappeared after bilateral carotid neurotomy. Anesthetic block of both carotid nerves produced transient reductions in tidal volume at any given temperature. We conclude that carotid body afferents contribute to the hyperventilation evoked by hyperthermia. After their interruption, such contribution is replaceable from other thermal afferents.

Effects of body temperature on chemosensory activity of the cat carotid body in situ

The effects of changes in body core temperature (TB) upon the frequency of chemosensory discharges (fx) from one carotid nerve were studied in pentobarbitone anesthetized cats. Raising TB from 35 to 40 degrees C increased fx in some cats, an effect more commonly seen after contralateral carotid neurotomy. In other animals, the simultaneously increased alveolar ventilation counteracted the above effect. A multiple correlation analysis of global data showed predicted increases in fx in response to raising TB at different CO2 levels.

How the carotid body works: Different strategies and preparations to solve different problems

This is a review of the different experimental approaches developed to solve the problems in our progress towards a comprehensive understanding of how arterial chemoreceptors operate. An analysis is performed oi the bases, advantages and limits of the following preparations: studies of ventilatory reflexes originated from carotid bodies (CBs) in the entire animal; recordings of CB chemosensory discharges in situ; CB preparations perfused in situ; CB explants in oculo; CB explants in ovo; CB preparations incubated in vitro; CB preparations superfused in vitro; CB preparations perfused and superfused in vitro: CB tissue slices in vitro; cells acutely dissociated from CBs; CB cells in tissue culture; petrosal ganglia superfused in vitro; petrosal ganglion cells in tissue culture; and co-cultures of CB and sensory ganglion cells. A brief historical account is given of the passage from one preparation to the next one. Emphasis is placed on personal experience with the different preparations whenever possible. Examples are given of the importance of selecting the appropriate experimental preparation for solving each particular theoretical problem. In fact, brilliant ideas on how the CB works have been unproductive until finding the adequate experimental approach to explore the validity of such ideas.

Ventilatory Chemosensory Drive in Cats, Rats and Guinea-pigs

Comroe and Schmidt (1938) proposed that decreasing PaO2is the effective stimulus for arterial chemoreceptors (aortic and carotid bodies), but they considered that these peripheral chemoreceptors could not play a role in the ventilatory control in normoxia. Nevertheless, electrophysiological recordings from the carotid (sinus) nerves showed chemoreceptor activity at normoxic PaO2, which became feeble or absent when the animal was made hyperoxic by breathing pure oxygen (Landgren and Zotterman, 1951; Astrand, 1954).