Mechanisms of low-glucose sensitivity in carotid body glomus cells.

OBJECTIVE: Glucose sensing is essential for the adaptive counterregulatory responses to hypoglycemia. We investigated the mechanisms underlying carotid body (CB) glomus cells activation by low glucose. RESEARCH DESIGN/METHODS AND RESULTS: Removal of extracellular glucose elicited a cell secretory response, abolished by blockade of plasma membrane Ca(2+) channels, and a reversible increase in cytosolic Ca(2+) concentration. These data indicated that glucopenia induces transmembrane Ca(2+) influx and transmitter secretion. In patch-clamped glomus cells, exposure to low glucose resulted in inhibition of macroscopic outward K(+) currents and in the generation of a depolarizing receptor potential (DRP). The DRP was abolished upon removal of extracellular Na(+). The membrane-permeable 1-oleoyl-2-acetyl-sn-glycerol induced inward currents of similar characteristics as the current triggered by glucose deficiency. The functional and pharmacological analyses suggest that low glucose activates background cationic Na(+)-permeant channels, possibly of the transient receptor potential C subtype. Rotenone, a drug that occludes glomus cell sensitivity to hypoxia, did not abolish responsiveness to low glucose. The association of Glut2 and glucokinase, characteristic of some high glucose-sensing cells, did not seem to be needed for low glucose detection. CONCLUSIONS: Altogether, these data support the view that the CB is a multimodal chemoreceptor with a physiological role in glucose homeostasis.

Catecholamines in paraganglia associated with the hepatic branch of the vagus nerve: effects of 6-hydroxydopamine and reserpine.

Paraganglia are clusters of cells containing catecholamines (CA), mainly norepinephrine (NE) and dopamine (DA). The presence of epinephrine (E), on the other hand, has only been determined by indirect methods in retroperitoneal paraganglia of newborn and aged rats. Because their location, paraganglia associated with the hepatic branch of the vagus nerve may be a possible source of CA for the liver. The main purposes of the present study were to determine CA levels and whether E can be found in the omentum minus which includes paraganglia associated with the hepatic branch of the vagus nerve, and then to study the effects of 6-hydroxydopamine and reserpine on their CA content. Twenty-four female Wistar rats were randomly ascribed to three groups receiving two intraperitoneal injections of either 6-hydroxydopamine, reserpine or saline. Twenty-four hours after the last administration the rats were anesthetized and a portion of the omentum minus was obtained. Left adrenal medulla and a liver fragment were also collected as controls. The samples were processed to be analyzed by high performance liquid chromatography and catecholamine histofluorescence. The results confirm previous reports about the presence of considerable amounts of norepinephrine and dopamine in paraganglia. Norepinephrine and dopamine in the omentum like the adrenal medulla were significantly depleted by reserpine but not by 6-hydroxydopamine treatment, suggesting that some other sources in addition to sympathetic terminals are responsible for CA in the omentum. On the contrary, both drugs reduced liver NE, consistent with the localization of this amine mainly to hepatic sympathetic terminals. Histofluorescence of the omentum revealed 2-4 paraganglia per tissue fragment. Paraganglia associated with the hepatic branch of the vagus nerve contain also E. The presence of perihepatic sources of extra-adrenal CA, and more specifically E, could be of physiological significance.

Chemosensitivity, plasticity, and functional heterogeneity of paraganglionic cells in the rat coeliac-superior mesenteric complex.

Chemosensitivity and plasticity of paraganglionic cells in the rat coeliac-superior mesenteric complex (CSMC) were investigated at a basal state of normoxia (21% O2) and after long-term moderate hypoxia (10% O2, 14 days). Chemical sympathectomy previous to hypoxia was performed to destroy principal ganglionic neurons and thus to allow measurement of the norepinephrine and dopamine content of paraganglionic cells. At the basal state, the CSMC contained dopaminergic (TH+/DBH-) and noradrenergic (TH+/DBH+) paraganglionic cells, the majority being of the noradrenergic type. After 14 days of hypoxia, this ratio was reversed and dopaminergic cells predominated, as indicated by a twofold increase of TH+ cells and a twofold decrease of DBH+ cells. Biochemically, hypoxia produced an increase in the content (1.6-fold) and utilization (1.4-fold) of dopamine as well as a smaller increase in the content of norepinephrine, with no change in its utilization rate. The dopaminergic activation induced by hypoxia persisted after sympathectomy with guanethidine. It is concluded that paraganglionic cells in the CSMC display a chemosensitive function. Furthermore, our findings indicate that paraganglionic cells are differentially affected by hypoxia, depending on their distribution and the nature of their neuromodulators. The alterations induced by hypoxia point out the phenotypic plasticity developed by paraganglionic cells in adaptation to hypoxia and further demonstrate the functional heterogeneity of this autonomic cell population in the rat CSMC.

Immunohistochemistry of small intensely fluorescent (SIF) cells and of SIF cell-associated nerve fibers in the rat superior cervical ganglion.

Double-labelling immunofluorescence was applied on single sections of the rat superior cervical ganglion to evaluate neurochemistry and connectivity of intraganglionic SIF cells. The synaptic vesicle membrane protein synaptophysin and secretoneurin, a newly discovered neuropeptide derived from secretogranin II, proved reliable molecular markers of this cell type, whereas serotonin and tyrosine hydroxylase immunoreactivities were observed in slightly incongruent SIF cell subpopulations. Immunolabelling for vasoactive intestinal polypeptide and neuropeptide Y occurred in few SIF cells. None of the above immunoreactivities were visibly altered by preganglionic or postganglionic denervation, while some SIF cells were immunolabelled for galanin or for the neuronal microtubule-associated protein MAP2 after postganglionic denervation. SIF cells were nonreactive for the pan-neuronal marker protein gene product (PGP) 9.5 or neurofilament 160 kD. Intense staining of NADPH-diaphorase in some SIF cells, suggesting catalytic activity of nitric oxide synthase, could not be substantiated by immunoreactivity for this enzyme. SIF cells were approached by nonidentical fiber populations immunoreactive for PGP 9.5, neurofilament, or neuropeptide Y, whereas immunoreactivities for galanin and vasoactive intestinal polypeptide were colocalized in fiber meshes around SIF cells. The findings indicate (1) neurochemical SIF cell heterogeneity, (2) SIF cell plasticity in response to ganglionic perturbation, and (3) a differentiated innervation of SIF cells in the rat superior cervical ganglion.

The glomus tympanicum: a middle ear chemoreceptor?

Negative pressure in the middle ear has been identified as an important factor in the pathogenesis of middle ear disease. Unfortunately, to date, the physiological mechanisms that control middle ear pressure are poorly understood. Recent interest has focused on the possible role of carotid body-like tissue in the middle ear (the glomus tympanicum) as a chemoreceptor for changes in the gas composition of the middle ear. From the few anatomical studies of normal glomus tissue in the temporal bone it would seem that glomus bodies may not be consistently present in the middle ear. In this study, glomus bodies were sought in histological preparations of the promontory mucosa and tympanic plexus. It was found that the glomus tympanicum is only occasionally present in the middle ear mucosa, and thus a chemoreceptor role specific to the middle ear is unlikely.

Paraganglia of the human recurrent laryngeal nerve.

Recurrent laryngeal nerves (RLNs) from human autopsy material were collected and prepared for light microscopy. Within five of eight investigated RLNs, paraganglia-like organs were found, the mean cross-sectional area of which was estimated to be 0.03 mm2. These organs were composed of cells resembling the type I and type II cells of the carotid body. The possible function of laryngeal nerve paraganglia is discussed.

Adrenergic mechanisms in oxygen chemoreception in the cat aortic body.

Sixteen cats were studied to test the hypothesis that oxygen chemoreception in the cat aortic body is dependent on the beta-adrenergic mechanism. The chemoreceptor activity was measured from a few aortic chemoreceptor afferents in each cat, anesthetized with alpha-chloralose (60 mg X kg-1). Three types of experiments were conducted. Aortic chemoreceptor responses to steady-state hypoxia (PaO2 range, 100-30 Torr) were measured (a) before and during intravenous infusion of the beta-receptor agonist, isoproterenol (0.5 micrograms X kg-1) in nine spontaneously breathing cats, and (b) before and after intravenous injection of the beta-receptor antagonist, propranolol (1 mg X kg-1) in seven cats which were paralyzed and artificially ventilated. In the third category (c) the stimulatory effect of hypotension on aortic chemoreceptor activity was measured in six of the seven cats in group (b) before and after propranolol injection. Isoproterenol infusion only moderately stimulated aortic chemoreceptor activity. This stimulation was blocked by propranolol. However, propranolol did not attenuate aortic chemoreceptor responses to hypoxia or to hypotension. We conclude that the beta-receptor adrenergic mechanism does not mediate oxygen chemoreception in the cat aortic body.

Effects of stimulation of aortic chemoreceptors on abdominal vascular resistance and capacitance in anaesthetized dogs.

1. Dogs were anaesthetized with chloralose, ventilated artificially, and the regions of the aortic arch and carotid sinuses were isolated vascularly and perfused with blood. The abdominal circulation was isolated vascularly, perfused at constant flow and drained from the inferior vena cava at constant venous pressure. Changes in vascular resistance were determined by calculating changes in abdominal aortic perfusion pressure, and changes in capacitance by integrating the changes in venous outflow. 2. Stimulation of aortic body chemoreceptors, either by changing the aortic arch perfusate from arterial to venous blood at constant perfusion pressure or by injection of sodium cyanide into the aortic arch, resulted in an increase in abdominal vascular resistance and a decrease in abdominal vascular capacitance. 3. After both cervical vagosympathetic trunks had been cut, stimulation of aortic chemoreceptors no longer resulted in resistance or capacitance responses. 4. These results indicate that stimulation of aortic chemoreceptors, like carotid chemoreceptors, results in reflex constriction of both resistance and capacitance vessels in the abdominal circulation.

Neural responses of the cat carotid and aortic bodies to hypercapnia and hypoxia.

The response (imp . s-1) of single- or few-fiber preparations from the carotid body (10 experiments) and the aortic body (5 experiments) to various levels of hypercapnia on different backgrounds of hypoxia were analyzed by two statistical techniques--analysis of variance and the Duncan's new multiple-range test. These analyses showed an initial statistically significant increase in the slope of the response to increasing arterial pressure of CO2 (PaCO2) as PaO2 fell. But the slope of the response to carbon dioxide later showed a clear tendency to become less; i.e., no significant increase in imp . s-1 when a PaCO2 rose (substantially) with normoxic (carotid body) and hypoxic (carotid and aortic bodies) backgrounds. The response of the aortic body to hypercapnia showed no statistically significant increase if the background was hyperoxia or normoxia. The characteristic of the chemoreceptor to become saturated in its response to carbon dioxide while still retaining its ability to respond to hypoxia suggests the possibility that at least some of the mechanisms involved in the chemoreception of hypoxia differ from those involved in the chemoreception of hypercapnia.

Arterial chemoreceptor-like activity in the abdominal vagus of the rat.

1. Centripetal activity in fibres in the ventral abdominal vagus nerve of the rat has been studied by recording from fine strands of the divided nerve within the abdomen. 2. In the starved animal, few spontaneously active fibres were located. A proportion of these, however, showed changes in activity in response to changes in F1 oxygen which were typical of arterial chemoreceptor afferent nerves. The resting discharge in these preparations was 0.8-8.0 impulses/sec. In response to extreme hypoxic hypoxia, histotoxic hypoxia or acetylcholine, this discharge increased markedly, with a maximum mean activity of up to 25 impulses/sec. 3. Both the mean/S.D. ratio and statistical comparison with a 'noise' equation were used to assess the apparent random nature of the spike intervals. The former indicated that the spike intervals were random but the latter test was inconclusive. 4. We suggest that this chemoreceptor-like activity originates from the abdominal vagal paraganglia and that these structures may be part of a more generally distributed chemoreceptor system.

Relative responses of aortic body and carotid body chemoreceptors to carboxyhemoglobinemia.

The effects of carbon monoxide inhalation and of consequent carboxyhemoglobinemia (HbCO) on the discharge rates of aortic body and carotid body chemoreceptor afferents were investigated in 18 anesthetized cats. In 10 experiments both aortic and carotid chemoreceptor activities were monitored simultaneously. Carbon monoxide inhalation during normoxia always stimulated aortic chemoreceptors before carotid chemoreceptors, and the steady-state response of aortic chemoreceptors to HbCO was greater than that of most carotid chemoreceptors. Only 2 of the 18 carotid chemoreceptor fibers tested showed a distinct increase in activity in response to moderate increases in HbCO%. Thus, oxyhemoglobin contributed substantially to maintain tissue PO2 of all aortic chemoreceptors and of a few carotid chemoreceptors. Hyperoxia diminished the response of both aortic and carotid chemoreceptors to HbCO, indicating a lowered tissue PO2 as the stimulus source. We hypothesize that the aortic bodies have a much lower perfusion relative to their O2 utilization compared to the carotid bodies. As a consequence, the aortic chemoreceptors are able to act as a sensitive monitor of O2 delivery and to generate a circulatory chemoreflex for O2 homeostasis. carotid chemoreceptors monitor O2 tension and initiate strong reflex effects on the level of ventilation.

Responses of the heart to stimulation of aortic body chemoreceptors in dogs.

We stimulated the aortic chemoreceptors in dogs that were anesthetized with chloralose and artificially ventilated by perfusing the isolated aortic arch with venous blood. Inotropic responses were determined by measuring the maximum rate of change of left ventricular pressure (dP/dt max) with aortic pressure and heart rate held constant. Stimulation of the aortic chemoreceptors resulted in an average increase in heart rate of 14 +/- 2.0 beats/min (mean +/- SE) from 166 +/- 7.7 beats/min and an increase in dP/dt max of 501 +/- 85 mm Hg/sec from 3508 +/- 154 mm Hg/sec. These changes were statistically significant (P less than 0.001). The afferent pathway of the reflex was shown to be in the vagus nerves and the efferent pathway in the cardiac sympathetic nerves. In some of the dogs, the carotid chemoreceptors were also stimulated. This resulted in decreases in heart rate and dP/dt max of 48 +/- 24 beats/min and 795 +/- 142 mm Hg/sec. Thus we have shown that stimulation of aortic chemoreceptors evokes chronotropic and inotropic responses opposite to those evoked from stimulation of carotid chemoreceptors.

Aortic body chemoreceptor responses to changes in PCO2 and PO2 in the cat.

Responses of aortic chemoreceptor afferents to a range of arterial carbon dioxide tension (Paco2) changes at various levels of arterial oxygen tension (Pao2) were investigated in 18 cats anesthetized with alpha-chloralose and maintained at 38 degrees C. Aortic chemoreceptor activity, end-tidal oxygen pressure, end-tidal carbon dioxide pressure, and arterial blood pressure were continuously monitored. Arterial blood gases were measured in steady states. Single or a few clearly identifiable afferents were studied during changes and steady states of Pao2 and Paco2. All the aortic chemoreceptor afferent discharge rates increased with Paco2 increases from hypercapnia (10-15 Torr) to normocapnia and moderate hypercapnia (30-50 Torr) and with Pao2 decreases from above 400 to 30 Torr. Hypoxia augmented the response to Paco2 most effectively in the range of 10-40 Torr. At any Pao2, the discharge rate reached a plateau with sufficient intensity of hypercapnia. The Paco2 stimulus threshold at a Pao2 of 440 Torr was about 15 Torr, and at a Pao2 of 60 Torr it was 10 Torr. In the transition from hypocapnia to hypercapnia, responses increased gradually, usually without an overshoot. The steady-state responses to Paco2 of the majority of aortic chemoreceptors resembled those of carotid chemoreceptors. The responses of both receptors can be attributed to the same basic type of mechanism.

Ventilatory control in peripheral chemoreceptor-denervated ponies during chronic hypoxemia.

The present study was designed to provide further insight into the role of the carotid and aortic chemoreceptors in ventilatory (VE) acclimatization during sojourn at altitude. Measurements were made: 1) on 10 ponies near sea level (SL, 740 Torr) under normal conditions, 2) on 6 of these at SL following chemoreceptor denervation (CD), and 3) subsequently on all 10 during 4 days of hypobaric hypoxia (PaO2 = 40-47 Torr). CD resulteo in hypoventilation at SL (deltaPaCO2 = d8 Torr, P less than 0.05), and it prevented hyperventilation normally observed with injection of NaCN and acute exposure to hypoxia (less than 1 h). In contrast, hyperventilation was evident in normal ponies during acute hypoxia (deltaPaCO2 = -6.7 Torr). Ventilation increased in both groups between the 2nd and 8th h of hypoxia (deltaPaCO2 from 1 h = -4 Torr, P less than 0.05). This change, a common characteristic of acclimatization, persisted throughout 4 days of hypoxia in the normal ponies. However, in the CD ponies this change was evident consistently only through the 12th h and after the 44 h hyperventilation was no longer evident. We conclude that the peripheral chemoreceptors are essential in ponies for normal VE acclimatization to this degree of hypoxemia. Two additional findings in CD ponies suggest the presence of a CNS inhibitory influence on the VE control center during chronic hypoxemia. First, acute hyperoxygenation on the 4th day of hypoxemia induced hyperventilation (deltaPaCO2 = -5 Torr, P less than 0.05). Second, again on the 4th day and during hyperoxygenation, VE responsiveness to CO2 and doxapram HCl was greater than at sea level.

Effects of sympathetic stimulation on carotid and aortic baroreceptors in the cat.

Variations in the discharge of baroreceptor units in the left aortic nerve were investigated during stimulation of the stellate ganglion both in the intact cat and in vitro perfusion of the aortic arch. The effects of stimulation of the peripheral cervical sympathetic trunk of the baroreceptor discharge in the carotid nerve during in vitro perfusion of the carotid artery were further studied. Stimulation of the stellate ganglion or the aortic nerve in the intact cat caused a simultaneous increase in arterial pressure, heart rate and number of baroreceptor impulses in a filament to the left aortic nerve. In the in vitro studies, decreases in the number of baroreceptor impulses both in the aortic and carotid nerves were produced in most cases during the stimulation of the sympathetic nerves. These effects were only observed during low pressure perfusion of the isolated artery and in low frequency changes. The infusion of norepinephrine caused a more marked decrease.

Chemoreceptor properties of glomus tissue found in the carotid region of the cat.

1. ;Miniglomera' appearing as small masses of tissue with ample vascularization were found around the common carotid artery of the cat. Physiological, gross anatomical and electron microscopic studies were conducted on these tissues.2. The chemosensory function of each ;miniglomus' was evident from the behaviour of the afferent nerve fibres supplying the tissue: afferent responses became more active during asphyxia, when the blood flow through the tissue was reduced or blocked and when cyanide or ACh were applied. The afferent impulses became more infrequent during hyperventilation.3. Sensory frequency response curves constructed against percentage of inhaled O(2) showed that the impulses of single units increased in frequency with lowering of O(2) content of the inhaled gas.4. These miniglomera are innervated by afferent fibres emerging from the nodose ganglion; sometimes these fibres are contained in the aortic or common carotid baroreceptor nerves, but sometimes they emerge as independent nerves. None of the miniglomera are supplied by branches of the sinus nerve.5. The fine structure of the miniglomus is similar to that of the carotid body. The tissue contains two types of cells: glomus cells which contain dense cored granules, and sustentacular cells whose fine processes enclose the former. Membrane densifications occur where glomus cells lie adjacent to one another or where they are contacted by nerve terminals. Nerve fibres are common in the miniglomus but they contact glomus cells less frequently than in the carotid body.