Chemosensory Discharge Research Articles

Contribution of in vivo microvascular PO2 in the cat carotid body chemotransduction.

To understand the interplay between microcirculatory control and carotid body (CB) function, we simultaneously measured carotid body microvascular PO2 (CBM PO2) and chemosensory activity in the cat in vivo under several experimental conditions. Cats were anesthetized with pentobarbital sodium, paralyzed, and artificially ventilated. CBs were exposed, and steady-state CBM PO2 was measured by the O2-dependent quenching of the phosphorescence of Pd-meso-tetra-(4-carboxyphenyl)porphine, which was administered intravenously. A few fibers of the carotid sinus nerve were used to record chemosensory discharges. At arterial PO2 (PaO2) of 103.4 +/- 4.1 Torr, CBM PO2 was 52.5 +/- 3.6 Torr (n = 9). Graded lowering of PaO2 from 160 to 50 Torr resulted in nearly proportional decreases in CBM PO2, but at lower PaO2 the decrease in CBM PO2 became more substantial. As PaO2 decreased, chemosensory discharge increased in parallel with CBM PO2. Hypercapnia and hypocapnia did not significantly change the relationship between PaO2 and CBM PO2, although the chemosensory discharge responded significantly. CBM PO2 and chemosensory discharge were not affected by hemorrhagic hypotension until arterial blood pressure fell below approximately 50 Torr and then CBM PO2 decreased and chemosensory discharge increased. The lack of a significant effect of hemorrhagic hypotension indicated that O2 delivery to CB was almost independent of the systemic blood pressure. Taken together, the observations suggest that CB microcirculation and PO2 are subject to control by intrinsic mechanisms and that CBM PO2 is compatible with oxidative metabolism playing a role in O2 chemoreception during hypoxia.

Differential modulation by extracellular ATP of carotid chemosensory responses.

The possibility that the carotid body has ATP surface receptors that mediate O2 chemoreception was tested. To distinguish between the event(s) initiating chemoreception and those at the neurotransmitter level, we also tested the chemosensory response to nicotine before and after ATP administration. Carotid bodies from cats anesthetized with pentobarbital sodium were perfused and superfused in vitro with modified Tyrode solution (PCO2 < 1 Torr, pH 7.4, 36 degrees C) equilibrated at PO2 > 400 or approximately 150 Torr while chemosensory discharge was recorded extracellularly. ATP and adenosine 5'-[gamma-thio]triphosphate stimulated discharge with similar dose dependence, whereas adenosine had little effect. ATP infusion for > or = 2 min evoked an initial stimulation of discharge followed by a decline to baseline (desensitization). Desensitization did not affect the response to hypoxia (perfusate flow interruption) but inhibited the response to nicotine (4-nmol pulse). Therefore, 1) the carotid body has surface ATP receptors that may mediate the chemosensory response to nicotine but not to hypoxia and 2) nicotinic receptors are not required for carotid body O2 chemoreception.

Effects of inorganic calcium channel blockers on carotid chemosensory responses in the cat.

The role of extracellular calcium entry through the voltage-operated calcium channels in response to hypoxia is controversial. Earlier observations (Eyzaguirre and Zapata 1965, Delpiano and Acker, 1989)are that upon removal or depletion of extracellular Ca2+ in vitro, the carotid chemosensory discharge increased during the first 20–40 min and then diminished. At this stage the responses to hypoxia and acetylcholine were also attenuated. More recently Shirahata and Fitzgerald (1991) reported that Ca2+removal in situ during hypoxia was promptly followed by a decrease in the chemosensory activity. Fidone et al.(1982) reported that incubation of carotid body in the Ca2+ free medium reduced dopamine release due to hypoxia. Working with freshly isolated glomus cells Biscoe and Duchen (1990), reported that removal of extracellular Ca2+ or use of calcium channel blocker did not prevent an increase in the intracellular Ca2+ due to hypoxia. Also, the effects of metabolic inhibitors on Ca2+ rise was not blocked by the foregoing procedures. Accordingly they concluded that the Ca2+ entry through the voltage-operated channels was not required for the hypoxic chemoreception. Besides, for the voltage-operated channels to participate in the hypoxic response, the cells needed to be depolarized first but there was no consistent depolarization. However, Sato et al. (1991) found evidence for Ca2+ increase in the glomus cells during hypoxia, thus Ca2+ controversy persisted. Previously we tested the effects of inorganic blockers of Ca2+ channel (Di Giulio et al. 1990) on carotid chemosensory activity and responses to hypoxia and found that Co2+stimulated chemosensory discharge and did not block the response to hypoxia. Recently we extended the study using the divalent cations Ni2+,Cd2+ and Mn2+ on the chemosensory discharges in the cat carotid body in vivo.

Thermal dependence of chemosensory activity in the carotid body superfused in vitro

We studied the relationship between chemosensory activity and temperature in carotid bodies excised from pentobarbitone-anesthetized cats, and superfused in vitro at flows between 0.4 and 2.0 ml/min with modified Tyrode's solution buffered with HEPES at pH 7.43. The basal frequencies of chemosensory discharges were recorded from the entire carotid nerve at different steady thermal conditions. For preparations superfused with saline equilibrated with 100% O 2, thermally dependent increases in frequency were observed, with significant differences between all nearby thermal stages separated by 0.5°C steps between 36.0 and 38.5°C. The larger gains were recorded between higher temperatures at high flows, between mid temperatures at intermediate flows, and between lower temperatures at low flows. The critical temperature for the calculated maximal gain was directly correlated to superfusion flow. The basal frequencies were consistently elevated when switching to saline equilibrated with 20% O 2 and no significant differences in mean ranks were recorded between 36 and 37°C, as between 38 and 39°C, but frequencies at 36–37°C were significantly higher than those at 38–39°C. Brief rises in chemosensory discharges were evoked by injections of NaCN applied to carotid bodies superfused with saline equilibrated with 100% O 2. The least effective dose was lower at 40°C than at 37.5° or 35.0°C, but the reactivity and slope were not significantly different. It is concluded that the carotid body chemoreceptors fulfill the criteria for being considered as thermosensors, and that their frequency of discharges is thermally modulated within a range close to physiological body temperature.

Effects of Doxapram on Carotid Chemoreceptor Activity in Newborn Kittens

Doxapram is a respiratory stimulant which acts on peripheral chemoreceptors and central respiratory neurons in a dose-dependent fashion in the adult cat. In the newborn, the mechanisms of action of doxapram are still unclear. To evaluate the effects of doxapram on the carotid chemosensory discharge and its relationship with dopaminergic mechanisms in the carotid body, two groups of kittens less than 13 days old, anesthetized, artificially ventilated and paralyzed, were prepared for the recording of a single or a few chemosensory afferents of the carotid sinus nerve. The chemosensory activity was recorded under five conditions of inspired gas mixtures (21 and 8% O2 in N2, 100% O2, 5 and 10% CO2 in O2). Group 1 (n = 9) received only doxapram and group 2 (n = 8) was pretreated with haloperidol (1 mg/kg), a dopamine D2-receptor blocker, before receiving doxapram. Doxapram significantly stimulated the discharge rate of the carotid chemoafferents under all conditions of inspired gas. The chemosensory discharge was increased by haloperidol, and was raised further after doxapram by an amount similar to group 1. For instance, in normoxia, the activity increased from 2.9 +/- 0.4 to 7.5 +/- 0.9 impulse/s (mean +/- SEM, p < 0.01) in group 1 and from 3.8 +/- 0.6 to 9.1 +/- 1.0 impulse/s (p < 0.01) in group 2. These results indicate that the mechanisms of response of carotid chemoreceptor to doxapram are developed in the newborn kitten and doxapram acts independent of the dopaminergic mechanisms in the carotid body.

Dual responses of carotid chemosensory afferents to dopamine in the newborn kitten

The effects of dopamine and of dopamine D2 receptor blocker haloperidol on the activity of carotid chemoreceptors were studied in 24 anesthetized, paralyzed and artificially ventilated newborn kittens aged 0–17 days. Single or few fiber preparations of chemoreceptors were made from one carotid sinus nerve. The responses of the chemosensory afferents to intravenous injections of dopamine (5–50 μg·kg −1) were studied in kittens breathing air and 8% O 2 in N 2. The effects of haloperidol on the chemosensory activity in air or 100% O 2 and on the chemosensory response to hypoxia were studied. Dopamine inhibited the chemosensory discharge in 25 44 studies in normoxia. Of the 9 studies performed in hypoxia, dopamine was excitatory in 5 or had no effect in 4 ( P<0.05 vs normoxia). Inhibition of the chemosensory discharge was observed in 24 37 studies performed in kittens aged more than 3 days and was predominantly excitatory in 6 7 studies in kittens aged 0–3 days ( P<0.01). One minute after haloperidol, the chemosensory discharge had increased significantly in all experiments. The biphasic pattern of chemosensory response to hypoxia (Marchal et al., Respir. Physiol. 87: 183–193, 1992) was not changed by haloperidol. The steady-state chemosensory activity was significantly increased after haloperidol, respectively from 3.9±0.7 impulses·sec −1 to 9.8±1.2 impulses·sec −1 in air, and from 13.1±1.4 impulses·sec −1 in hypoxia (mean ± SSEM, P<0.03). It is concluded that the dopaminergic mechanisms are active in the carotid body of the kitten, and the observed responses to dopamine and haloperidol are qualitatively similar to those reported in the adult cat.

Intracellular pH and oxygen chemoreception in the cat carotid body in vitro.

To test the hypothesis that O2 chemoreception in the carotid body (CB) is mediated by cellular acidosis, we simultaneously measured responses of the chemosensory and intracellular pH (pHi) to agents that are known to change pHi and studied the effects of hypoxia and ischemia on these variables in the cat CB. The CB was perfused and superfused in vitro with a modified Tyrode's solution at 36.0 +/- 0.5 degrees C with or without CO2-HCO3- (pH 7.40) and equilibrated at a given PO2. Chemosensory discharges were recorded from the whole carotid sinus nerve. To measure pHi changes, the CB was loaded with the pH-sensitive indicator 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, and the fluorescence (excitation 420-490 nm, emission greater than 515 nm) was detected by an intensified charged coupled device camera with an epifluorescence macroscope. Boluses of Tyrode's solution (0.5 ml, free of CO2-HCO3-) containing sodium acetate or NH4Cl prolonged perfusion of acid Tyrode's solution (pH 7.20-6.50), and boluses of Tyrode's solution with CO2-HCO3- were used. A decrease of fluorescence indicated pHi turning acid, and an increase of fluorescence indicated a change in alkaline pHi. Chemosensory activity varied inversely with the fluorescence change after application of these agents. Interruption of perfusate flow or application of hypoxic perfusate resulted in large increases in chemosensory discharge without any change in the fluorescence. The results indicated that chemosensory responses to brief ischemia and hypoxia were not mediated by a fall of pHi of CB cells, whereas those to CO2 and extracellular acidity were associated with decreases in pHi.

Optical measurements of oxygen and electrical measurements of oxygen chemoreception in the cat carotid body.

Measurements of oxygen pressures within the carotid body have been of particular importance for elucidating the mechanism(s) of oxygen chemoreception. An optical technique based on the quenching of phosphorescence by oxygen that is capable of imaging oxygen within the microvasculature has been developed (Rumsey et al, 1988, Wilson et al, 1991). These optical measurements are non-invasive and in particular have the advantage of determining oxygen pressures throughout the entire tissue. We have previously applied this technology to the study of the isolated perfused/superfused cat carotid body and oxygen chemoreception (Rumsey et al, 1991), relying primarily on imaging of phosphorescence intensity. Although the latter is quite useful for following dynamic changes in oxygen pressures, calculation of oxygen pressure is only qualitative. In addition, the carotid body was perfused and superfuised with cell-free media (Iturriaga et al, 1991). In the present report, we describe measurements of oxygen pressure determined from images in phosphorescence lifetimes (Wilson et al, 1991) in the cat carotid body using an in vivo preparation (Lahiri and DeLaney, 1975). These determinations were compared to those obtained previously with the in vitro preparation, thereby permitting comparison of the oxygenation of this organ in the presence and absence of blood, respectively. In both cases, chemosensory discharge was recorded in order to provide measurements of the expression of oxygen chemoreception.

Carotid body chemoreception in the absence and presence of CO 2[sbnd]HCO 3−

Carotid body (CB) chemosensory responses to natural and pharmacological stimuli were studied in vitro in the presence and nominal absence of CO 2 HCO 3 − in the perfusion-superfusion media. The CBs obtained from cats (n = 10), anesthetized with sodium pentobarbitone, were simultaneously perfused and superfused with a modified Tyrode solution at36.5 ± 0.5°C, equilibrated respectively with PO 2 of 120 and< 20 Torr. The Tyrode, nominally free of CO 2 HCO 3 − (HEPES-NaOH, pH 7.38, 310 mOsm), was used first. Subsequently the Tyrode containing HEPES-HCO 3 − equilibrated with PCO 2 of 36.8 Torr (pH 7.38) was used. Chemosensory discharges were recorded from the carotid Sinus nerve. Both hypoxia (PO 2 = 20–25Torr) and ischemic hypoxia stimulated the discharge in the absence and presence of CO 2 HCO 3 −. However, the presence of CO 2 HCO 3 − significantly raised the baseline activity, augmented the speed, sensitivity and the maximal responses to both types of hypoxia. Hypercapnic perfusate (PCO 2 = 65Torr at pH7.17) produced a peak response equally promptly in the absence and presence of CO 2 HCO 3 − in the ongoing perfusate at but generated a larger and more sustained response. Presence of CO 2 HCO 3 − strongly potentiated the responses to cyanide (10 −10-10 −7 mol) but less strikingly the responses to nicotine (10 −11-10 −8 mol). Thus, the extracellular CO 2 HCO 3 − significantly improved the response to hypoxia but was not essential for O 2 chemoreception. The underlying mechanisms of the effect of CO 2 HCO 3 − is likely to be mediated by the Cl − HCO 3 − anion exchanger in the pH regulation of glamous cells.

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.

Optical measurements of the dependence of chemoreception on oxygen pressure in the cat carotid body

The relationship between oxygen pressure (PO2) in the carotid body and carotid sinus nerve discharge was evaluated in the isolated perfused/superfused cat carotid body using the oxygen-dependent quenching of phosphorescence. Images of phosphorescence intensity arising from Pd-coproporphyrin within the microcirculation of the carotid body provided measurements of intravascular PO2. These measurements were substantiated by determining phosphorescence life-time. The carotid body was perfused in the isolated state via the common carotid artery with N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid-buffered Tyrode solution, pH 7.4, at a constant pressure of 80 mmHg. Superfusion was maintained with similar media equilibrated with 100% argon. PO2 in the exchange vessels was markedly less than that in the perfusate entering the carotid artery, 23 +/- 3 and 45 +/- 3 Torr for normoxic (111 +/- 15 Torr) and hyperoxic (345 +/- 72 Torr) perfusates, respectively. Chemosensory discharge rose slowly in response to a brief interruption of perfusate flow as PO2 steadily declined from either of these capillary PO2 values to approximately 10 Torr. Between approximately 10 and 3 Torr, chemosensory discharge increased strikingly, concomitant with an enhanced rate of oxygen disappearance, from -36 +/- 4 to -69 +/- 13 (92% change) and -28 +/- 3 to -48 +/- 3 (71% change) Torr/s for normoxic and hyperoxic perfusates, respectively. As PO2 fell below approximately 3 Torr, oxygen disappearance slowed and neural activity decayed. Thus the relationships between microvascular PO2 and chemosensory discharge and between oxygen disappearance and neural discharge suggest that oxygen metabolism in the carotid body determines the expression of oxygen chemoreception.

Carbonic anhydrase and chemoreception in the cat carotid body

To test the hypothesis that CO2 and O2 chemoreception in the carotid body (CB) may depend on its carbonic anhydrase (CA) activity, we used an in vitro cat CB preparation and studied the effects of methazolamide, a permeable CA inhibitor (pK 7.3), on the chemosensory responses to CO2, O2, and nicotine. The isolated CB was perfused and superfused with Tyrode solution, free of CO2-HCO3-, at 36.0 +/- 0.5 degrees C. The frequency of chemosensory discharges was recorded from the whole carotid sinus nerve. The responses to bolus injections (0.3-0.5 ml) of Tyrode solution equilibrated with PCO2 of 38-110 Torr, switching from HEPES to CO2-HCO3- Tyrode (PCO2 = 25-60 Torr) for about 3 min, hypoxic Tyrode (PO2 = 25-30 Torr) for 2-8 min, perfusate flow interruptions for approximately 4 min, and bolus injections of nicotine (4 nmol) were studied before, during, and after perfusion (30-45 min) with methazolamide (42.4 microM). Methazolamide reversibly inhibited, delayed, and reduced the responses to transient CO2 stimulus, diminished the onset of but not the late response to prolonged CO2 stimulus, and delayed but did not decrease the responses to hypoxia and perfusate interruption. The response to nicotine did not change. The results indicated that CA in the glomus cells played a crucial role primarily in the speed and magnitude of the initial response to CO2 stimulus and indirectly influenced O2 chemoreception. These effects were upstream from the nicotine receptor-mediated sensory response.

Carotid chemosensory timing effects on cervical sympathetic discharges in the cat

The hypothesis that respiratory phase-related sympathetic nerve activity would manifest timing effect of carotid chemosensory input with respect to the central respiratory drive was tested in the anesthetized, paralyzed and artificially ventilated cats which were also vagotomized and tracheostomized. Preganglionic cervical sympathetic nerve fibers (PSNF) which were clearly responsive to carotid chemosensory stimulation were selected for the test. Simultaneously with the PSNF, phrenic nerve (PN) and internal intercostal expiratory nerve (IICEN) activities were also recorded. In most instances, carotid chemosensory discharges were monitored in order to register the precise timing of its input to the brain-stem. Timed stimulation of carotid chemoreceptors was elicited by bolus injections of cyanide (10–20 μg), nicotine (10–20 μg) and CO 2-saturated saline (0.2–0.5 ml) into the base of the common carotid artery. Two types of PSNFs were identified: type I discharged only in synchrony with the PN activity ( 13 47 ) and type II fired independently of, but entrained to, PN activity ( 34 47 ). Type I displayed characteristic timing effects of carotid chemoreflex on PN discharges. However, the sympathetic activity did not share the after-discharge of PN which persisted beyond the carotid chemoreceptor stimulation. Type II did not manifest any timing effect of carotid chemoflex with respect to PN and IICEN activities. These results suggest the following model of carotid chemoreflex effects on sympathetic neuron activities: a group is exclusively gated by the properties of central respiratory drive whereas the other is not gated but entrained by it in the spontaneously breathing anesthetized cat.

In vitro perfused-superfused cat carotid body for physiological and pharmacological studies

An in vitro perfused carotid body preparation was developed to study its chemosensory responses to physiological and pharmacological stimuli. The carotid bifurcation with the carotid body was vascularly isolated and excised from pentobarbital sodium-anesthetized cats. The CB was perfused in a chamber by gravity (80 Torr) with modified Tyrode's solution (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid-NaOH at pH 7.40) equilibrated at a given Po2 and superfused with the same medium at (Po2 of 20 Torr). The temperature was maintained at 35.5 +/- 0.5 degrees C. The frequency of chemosensory discharges (CD) was recorded from the whole carotid sinus nerve (n = 24), and the responses were tested by repeated interruptions of perfusate flow (SF), perfusion with hypoxic medium, and injections of nicotine and cyanide (0.1 nmol to 1 mumol) and hypercapnic medium. During hyperoxic perfusion, SF resulted in a sigmoidal increase in CD, reaching a maximum that was 23.6 +/- 4.4-fold greater than the basal activity. Restoration of flow returned CD promptly to basal values. After normoxic perfusion, SF led to a similar maximal activity more rapidly, but the duration was shorter. Reduction of the perfusate PO2 (Po2 from 450 Torr to 150, 30, and less than 10 Torr) caused a nonlinear increase in CD. CO2 stimuli (PCo2 38-110 Torr) resulted in a linear increase in CD. Nicotine or cyanide increased CD in a dose-dependent manner. The preparation retained its initial responsiveness for 2-3 h, making extensive experimental studies feasible.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of [125I]Endothelin Binding Sites in the Region of the Carotid Bifurcation and Brainstem of the Cat

We have demonstrated by autoradiography, displaceable binding for [125I]endothelin-1 ([125I]ET-1), [125I]endothelin-2 ([125I]ET-2), and [125I]endothelin-3 ([125I]ET-3) in the cat carotid bifurcation as well as in the nucleus of the tractus solitarius, where baroreceptor and chemoreceptor afferents from the carotid body and sinus terminate. There was also significant binding in the nodose and superior cervical ganglia. Barosensory and chemosensory discharge was recorded from filaments of the carotid sinus nerve in cats anesthetized with pentobarbitone. Intra-carotid injection of ET-1 or ET-3 (4-402 pmoles) caused transient dose-related depression of baroreceptor discharge without any immediate effects on systemic blood pressure (BP) or heart rate; there was a delayed biphasic effect on BP. ET-1 had little effect on chemosensory discharge during the first 15 s post-injection, but there was a delayed (45-90 s) dose-related increase in discharge. The effects of all three ETs were qualitatively similar, and ET enhanced chemoexcitation evoked by either acetylcholine or sodium cyanide. Our results show that (a) ET binding sites are located in the baroreceptor and chemoreceptor afferent pathways and (b) ETs can influence afferent activity of baroreceptors and chemoreceptors. Further studies are needed to determine the significance of these findings, particularly with regard to reflex control of the cardiovascular system.

Arterial chemoreceptor involvement in salicylate-induced hyperventilation in rats.

1. The extent to which peripheral arterial chemoreceptors are involved in the respiratory stimulant action of salicylates has been investigated in rats. 2. Injection of sodium salicylate (200 mg kg-1, single dose i.v.) caused a rapid transient hyperventilation that was not obtained when the carotid chemoreceptors were denervated by section of the carotid sinus nerves. A delayed (10 min) increase in respiration occurred regardless of whether or not the carotid nerves were sectioned. 3. Intravenous infusions of sodium salicylate (0.5 or 4 mg kg-1 min-1) caused hyperventilation in barbiturate-anaesthetized rats. The threshold dose for respiratory stimulation was significantly lower when the carotid sinus nerves were intact than when they were bilaterally sectioned, and the same pattern was observed following intravenous injections of sodium salicylate (cumulative doses) in anaesthetized and conscious rats. 4. Bilateral sectioning of the vagosympathetic nerve trunks did not significantly affect hyperventilation evoked by salicylate, suggesting that this response does not involve actions of salicylate on sensory receptors innervated by these nerves. 5. Administration of salicylate close-arterial to a carotid body, by local perfusion or cross-perfusion of a carotid sinus, led to an increase in respiration when the ipsilateral carotid nerve was intact, but not when it was sectioned. 6. Neuropharmacological studies on anaesthetized rats showed that chemosensory discharge, recorded from a sectioned carotid nerve, increased in response to salicylate injections with a similar dose-response pattern to the hyperventilation. Salicylate had no effect on baroreceptor discharge. 7. We conclude from our experiments that arterial chemoreceptors do contribute to salicylate-induced hyperventilation, and are almost exclusively responsible for the initial phase of the response in rats. Later increases in breathing are independent of reflexes from arterial chemoreceptors and result from actions at other sites, including the CNS. The therapeutic implications of our results are discussed.

Ganglioglomerular nerves influence responsiveness of cat carotid body chemoreceptors to almitrine

A bolus injection of almitrine bismesylate (0.5 mg · kg −1 i.v.) in anaesthetised artificially ventilated cats caused a significantly greater increase in carotid chemosensory discharge in animals with sectioned ipsilateral ganglioglomerular sympathetic nerves in comparison with a group in which these nerves were intact. Plasma levels of almitrine were similar in both groups. Responses to hypoxia and hypercapnia post-almitrine were also bigger if the ganglioglomerular nerves were cut. Domperidone (10–50 μg · kg −1 i.a), a dopamine D 2 receptor antagonist, greatly increased the responsiveness of chemoreceptors to almitrine in ganglioglomerular nerve-intact preparations. Almitrine-induced chemosensory activity was unaffected by illuminating the carotid bifurcation with light from a fibre optic lamp, regardless of whether or not the ganglioglomerular nerves were cut. It is concluded that almitrine may directly or indirectly activate an efferent pathway in the ganglioglomerular nerves to cause depression of chemoreceptor activity, possibly by releasing dopamine to act at D 2 dopamine receptors in the carotid body.

Flow-dependent chemosensory activity in the carotid body superfused in vitro

The relationship between carotid body chemoreceptor activity and flow was studied in preparations superfused in vitro. The carotid bodies were excised from pentobarbitone-anesthetized cats and superfused with modified Tyrode's solution, buffered with HEPES-NaOH to pH 7.41. The bath temperature was kept constant at 37.7°C. The frequency of chemosensory discharges from the entire carotid nerve was determined during steady-state superfusion with 100% or 20% O 2-equilibrated saline at flow rates between 0.15 and 2.95 ml/min, and during 5 min flow interruptions. The peak frequency evoked by flow interruptions was maximal and independent of previous superfusion flows, but the half-excitation time of chemosensory responses to flow interruption was minimal when preceded by superfusion with 100% O 2-equilibrated saline at 0.7 ml/min. In steady-state conditions, mean chemosensory activity was higher at lower rates of flow, and, at constant flow, higher under 20% O 2 than under 100% O 2. To allow comparisons of all data, basal frequencies at given basal flows were referred to their own maximal frequencies evoked by flow interruptions. The best fitting for the relation between basal chemosensory activity and superfusion flow was provided by inverse sigmoid (logistic decay) curves; r = −0.90and−0.84, at 100% and 20% O 2 levels, respectively. The maximal gains were at about 0.78 and 0.86 ml/min, respectively. It is concluded that the chemosensory discharge frequency recorded from carotid bodies superfused in vitro is determined by the superfusion flow, when all other natural chemoreceptor stimuli are held constant.

Characterization of opioid receptors in the cat carotid body involved in chemosensory depression in vivo.

The effects of selective opioid receptor agonists and antagonists on neural discharge recorded from carotid body arterial chemoreceptors in vivo were studied in anaesthetized cats. Mean ID50 values were determined for each agonist and used to assess chemodepressant potency on intracarotid (i.c.) injection in animals artificially ventilated with air. [Met]enkephalin, [Leu]enkephalin, [D-Ala2, D-Leu5]enkephalin and [D-Pen2, D-Pen5]enkephalin were more potent chemodepressants than [D-Ala2, Me-Phe4, Gly-ol5]enkephalin, dynorphin (1-8) or ethylketocyclazocine; morphiceptin (mu-agonist) was inactive. The rank order of potency was compatible with the involvement of delta-opioid receptors in opioid-induced depression of chemosensory discharge. ICI 154129, a delta-opioid receptor antagonist, was used in fairly high doses and caused reversible dose-related antagonism of chemodepression induced by [Met]enkephalin. It also antagonized depression caused by single doses of [Leu]enkephalin, [D-Ala2, D-Leu5]enkephalin, [D-Ala2, Me-Phe4, Gly-ol5]enkephalin or dynorphin (1-8). ICI 174864, a more potent and selective delta-opioid receptor antagonist, also antagonized chemodepression induced by [Met]enkephalin or by the selective delta-receptor agonist [D-Pen2, D-Pen5]enkephalin. Comparison of background or 'spontaneous' chemosensory discharge during the 30 min periods immediately before and after injecting ICI 174864 (0.1-0.2 mg kg-1 i.c.) showed a significant increase in discharge in one experiment, but in four others discharge was either unaffected or decreased after the antagonist, which argues against a toxic depression of chemosensors by endogenous opioids under resting conditions in our preparation. Sensitivity of the carotid chemoreceptors to hypoxia (ventilating with 10% O2) was increased significantly after ICI 174864, which could be taken as evidence that endogenous opioids depress chemosensitivity during hypoxia. In contrast, responsiveness to hypercapnia was reduced after the antagonist, implying that endogenous opioids may potentiate chemoreceptor sensitivity during hypercapnia. The results obtained using 'selective' agonists and antagonists provide evidence that depression of chemosensory discharge caused by injected opioids involves a delta type of opioid receptor within the cat carotid body. Endogenous opioids may modulate arterial chemoreceptor sensitivity to physiological stimuli such as hypoxia and hypercapnia.

Pharmacological characterization of the receptor involved in chemoexcitation induced by adenosine.

Experiments were performed on cats anaesthetized with pentobarbitone in which carotid body chemoreceptor activity was recorded from the peripheral end of a sectioned carotid nerve. Intracarotid (i.c.) injections of adenosine and its analogues, NECA (5'-N-ethylcarboxamidoadenosine), L-PIA(L-N6-phenylisopropyladenosine), and D-PIA(D-N6-phenylisopropyladenosine), caused dose-related increases in chemosensory discharge. The rank order of potency as chemoreceptor stimulants was: NECA greater than adenosine greater than L-PIA greater than D-PIA. Infusion of theophylline antagonized the chemoexcitatory effects of NECA, and infusion of 8-phenyltheophylline (8-PT), which is a more potent adenosine antagonist with less activity as a phosphodiesterase inhibitor, reduced the chemoexcitation induced by adenosine. Infusion of 8-PT (10 micrograms min-1 i.c.), a dose which substantially reduced the effect of injected adenosine, also reduced the sensitivity of carotid chemoreceptors to hypoxia (10% O2 for 4 min). It is concluded that the adenosine receptors in the cat carotid body which mediate chemosensory excitation are xanthine-sensitive and appear to be of the A2 sub-type. Adenosine, released within the carotid body by physiological stimuli, may be involved in chemoexcitation.