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.

Long-term hypoxia induces changes in the substance P immunoreactivity pattern in laryngeal nerve paraganglia.

We previously showed that long-term hypoxia increases the dopamine content in rat laryngeal nerve paraganglia. In the present study paraganglia of rats exposed to hypoxia (10 +/- 0.5% O2) for 14 days were examined immunohistochemically to detect changes in the expression of neuropeptides and catecholamine-synthesizing enzymes. Hypoxia induced an intense cellular substance P (SP)-like immunoreactivity (LI) in some paraganglia and an increase in the number of stromal nerve fibers showing SP-LI in others. The patterns of tyrosine hydroxylase-, dopamine-beta-hydroxylase-, phenylethanolamine-N-methyltransferase-, vasoactive intestinal polypeptide-, neuropeptide-Y and calcitonin gene-related peptide-LI were not changed in response to hypoxia. The results show that hypoxia induces changes in the pattern of SP immunoreactivity in laryngeal nerve paraganglia and may indicate that SP plays a role in the regulation of catecholamine metabolism in this tissue.

Studies of local anesthetic action on natural spike activity in the aortic nerve of cats.

The aortic nerve was used to study the blocking action of procaine and bupivacaine on natural spike activity. In anesthetized cats, a segment of the aortic nerve was placed in a perfusion chamber and exposed to increasing drug concentrations, varying pH, while temperature remained constant. Total nerve activity was recorded continuously, and its change was related to drug concentration. The half-time of recovery following drug wash-out was also determined. At pH 7.4, the minimal blocking concentration was 0.5 X 10(-3) mol/l for procaine and 0.05 X 10(-3) mol/l for bupivacaine, the half-times of recovery 1.4 and 3.0 min, respectively. Procaine and bupivacaine reversibly blocked natural spike activity at the same concentrations as they blocked electrically evoked activity. The aortic nerve, whose physiologic spike traffic can be followed continuously for hours, may be used to advantage for studying the long-term effects of local anesthetics in vivo.

Aortic body chemoreceptor responses to dopamine, haloperidol, and pargyline.

Aortic chemoreceptor activity, from single- or few-fiber afferent nerve preparations, was measured in response to dopamine and a dopaminergic blocker, haloperidol, in 18 anesthetized cats. In six of these cats the effect of dopamine was assessed before and after inhibiting monoamine oxidase (MAO) by pargyline. Intravenous dopamine infusion (7-14 microgram X kg-1 X min-1) had a generally inhibitory effect on aortic chemoreceptor activity, but the magnitude of this effect varied with arterial partial pressure of O2 (Pao2) levels. The inhibitory effect of dopamine increased as Pao2 levels fell, and at severely hypoxic Pao2 levels (below 30 Torr) exogenous dopamine had no significant effect. The inhibitory effect of dopamine also increased during hyperoxic hypercapnia. Blockade of dopamine receptors in the aortic body by haloperidol-stimulated chemoreceptor activity significantly during hypoxia, suggesting an O2-dependent release of dopamine from the aortic body as Pao2 falls. Inhibition of MAO by pargyline had no significant effect on the control rate of activity at any level of Pao2 but augmented the inhibitory effect of exogenously administered dopamine. These data indicate that MAO is not significantly involved in the degradation of endogenous dopamine at the aortic receptor sites, but may participate in the degradation of exogenous dopamine.

Relative contribution of carotid and aortic bodies to cyanide-induced ventilatory responses in the cat.

The participation of afferences from carotid and aortic bodies to the hyperventilation caused by cytotoxic hypoxia was assessed in pentobarbitone-anesthetized cats. Dose-response curves for the ventilatory effects induced by i.v. injections of NaCN were obtained before and after successive denervations of peripheral chemoreceptors, in different sequences. Bilateral aortic neurotomy (BAN) or unilateral carotid neurotomy (UCN) did not affect significantly the minimal sensitivity to the drug, although maximal reactivity was reduced in some cats. After bilateral carotid neurotomy (BCN), with preservation of aortic nerves, sensitivity was reduced, but hyperventilation was still provoked by large doses of cyanide. BAN + BCN abolished the ventilatory responses to the drug. In cats with BAN + UCN, ventilatory responses had a high degree of correlation with increases of carotid chemosensory discharges in the range between ca 200% of control and the gasping threshold. It is concluded that the aortic bodies of the cat play a significant role in the hyperventilation produced by cytotoxic hypoxia, although it is less marked than that induced through the carotid bodies.

Long-lasting hyperventilation induced by almitrine: evidence for a specific effect on carotid and thoracic chemoreceptors.

In anesthetized dogs, almitrine (0.5-3 mg/kg i.v.) induced a dose-dependent increase in respiratory rate and ventilation. The aortic and carotid chemoreceptors were involved in the effects of almitrine. Section of both carotid sinus nerves and vagus nerves abolished the effects of the drug on respiration. The respiratory response did not occur in dogs with bilateral lesions of the nucleus of the solitary tract. The electrical activity of chemoreceptor fibres was increased. Perfusion of almitrine into the carotid artery stimulated respiration. Inhalation of pure oxygen shifted the dose-response curve of the respiratory effect towards the right. Almitrine slightly stimulated ventilation in dogs with bilateral section of carotid sinus nerves and aortic nerves and this disappeared when both vagus nerves were cut indicating that this effect was mediated through some chemoreceptor fibres present in the vagus nerves or through afferent vagal fibres.

[An extremely sensitive preparation for vasotocin action: the fresh-water eel aortic bulb]. / Une préparation particulièrement sensible à l'action de la vasotocine: le bulbe aortique de l'anguille d'eau douce

Transversal and longitudinal aortic bulb strips show isometric contractions in response to the vasotocin action. The extreme sensitivity of the preparations correspond to a final hormone concentration of 8.25 X 10(-16) M.

An electron microscopic study on the effects of reserpine on the subclavian glomera of the rabbit.

Young male and female New Zealand white rabbits were given a daily subcutaneous injection of reserpine (Serpasil, Ciba; 3 mg/kg) for two days and were sacrificed 24 hours after the last injection. The subclavian glomera (aortic bodies) were processed for electron microscopy to determine the effects of this biogenic amine depleting agent on the electron-opaque cytoplasmic granules of the parenchymal type I cells. Observations of glutaraldehyde-osmium tetroxide fixed glomera from reserpinized animals showed a slight decrease in granule density of the type I cells. Glomera fixed in glutaraldehyde and incubated in potassium dichromate (pH 4.1) demonstrated a reduction in granule opacity following reserpine treatment. Control glomera incubated in potassium dichromate displayed electron-opaque granules. These results indicate that reserpine does deplete the amines without granule disappearance or changes in granule population. The positive reaction of the control tissue granules to potassium dichromate incubation suggests that the predominant biogenic amines in the electron-opaque granules are unsubstituted monoamines. Persistence of the opaque granules following reserpinization and glutaraldehyde-osmium tetroxide double fixation, may be due to amine-binding protein within the granules. The mode of granule depletion could not be ascertained with certainty.