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A Obeso

Publications and source records attributed to A Obeso.

12 recordsLinked to original sources
PubMed1992

Oxygen and acid chemoreception in the carotid body chemoreceptors.

The carotid bodies are arterial chemoreceptors that are sensitive to blood PO2, PCO2 and pH. They are the origin of reflexes that are crucial for maintaining PCO2 and pH in the internal milieu and for adjusting the O2 supply according to the metabolic needs of the organism in situations of increased demand, such as exercise and while breathing at decreased O2 partial pressures during ascent or when living at high altitude. Chemoreceptor cells of the carotid body transduce the blood-borne stimuli into a neurosecretory response that is dependent on external Ca2+. These cells have an O2-sensitive K+ current that is reversibly inhibited by low PO2. It is proposed that the depolarization produced by inhibition of this K+ current activates Ca2+ channels; Ca2+ influx and neurosecretion follow. The cells have also a potent Na(+)-Ca2+ antiporter that could be responsible for the intracellular Ca2+ rise required to trigger the release of neurotransmitters during high PCO2 or low pH stimulation.

Acids↗
PubMed1992

The role of dihydropyridine-sensitive Ca2+ channels in stimulus-evoked catecholamine release from chemoreceptor cells of the carotid body.

The present study utilized an in vitro preparation of the rabbit carotid body, with tissue catecholamine stores labeled by incubation with 3H-tyrosine. The goal was to characterize pharmacologically the voltage-dependent Ca2+ channels present in the type I (glomus) cells of this arterial chemoreceptor organ, and to elucidate their role as pathways for Ca2+ entry. We found that release of 3H-dopamine induced by high external potassium was over 95% dependent on external calcium concentration and that this release was 90-100% inhibited by the dihydropyridine antagonists, nisoldipine and nitrendipine, and was potentiated by the dihydropyridine agonist, BayK 8644. Therefore, any stimulus-induced, calcium-dependent release of 3H-dopamine that was inhibited by nisoldipine and potentiated by BayK 8644, was considered to be supported by Ca2+ entry into the cells via voltage-dependent Ca2+ channels. Significant differences were observed in the release of 3H-dopamine induced by 75 vs 25 mM K+. On prolonged stimulation, release induced by 75 mM K+ was large and transient, whilst that induced by 25 mM K+, although more moderate, was sustained. The release elicited by 75 mM K+ was inhibited approximately 90% by 1.5 mM Co2+ or 625 nM nisoldipine, while release by 25 mM K+ was completely blocked by 0.6 mM Co2+ or 125 nM nisoldipine. Low PO2-induced release of 3H-dopamine was 95% dependent on Ca2+, and was inhibited by nisoldipine (625 nM) in a manner inversely proportional to the intensity of hypoxic stimulation, i.e. 79% inhibition at a PO2 of 49 Torr, and 20% inhibition at PO2 of 0 Torr. BayK 8644 potentiated the release induced by moderate hypoxic stimuli. Release elicited by high PCO2/low pH, or by Na(+)-propionate or dinitrophenol-containing solutions, was approximately 80% Ca(2+)-dependent, and the dihydropyridines failed to modify this release. It is concluded that type I cells possess voltage-dependent Ca2+ channels sensitive to the dihydropyridines, which in agreement with previous electrophysiological data should be defined as L-type Ca2+ channels. Calcium entry which supports the release of 3H-dopamine elicited by moderate hypoxia should occur mainly through these channels while the release induced by strong hypoxic stimuli will be served by Ca2+ entry which occurs in part via voltage-dependent Ca2+ channels, and in part through an additional pathway, probably a Na+/Ca2+ exchanger.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Effects of almitrine on the release of catecholamines from the rabbit carotid body in vitro.

1. Almitrine increases ventilation by stimulating the carotid body (CB) arterial chemoreceptors but neither its intraglomic target nor its mechanism of action have been elucidated. 2. We have tested the hypothesis that chemoreceptor cells are targets for almitrine by studying its effects on the release of 3H-catecholamines in an in vitro rabbit CB preparation. 3. It was found that almitrine (0.3 and 1.5 x 10(-6) M; i.e. 0.2 and 1 mg ml-1) increases the resting release of 3H-catecholamines from CBs (previously loaded with [3H]-tyrosine) incubated in a balanced 95% O2/5% CO2-equilibrated solution. 4. Almitrine at a concentration of 3 x 10(-6) M (2 mg l-1) also augmented the release of 3H-catecholamines elicited by incubating the CBs in a hypoxic solution (equilibrated with 7% O2/5% CO2 in N2), by high external K+ (35 mM) and by veratridine (2 x 10(-5) M), but did not modify release induced by dinitrophenol (7.5 x 10(-5) M). 5. At the same concentration (3 x 10(-6) M), almitrine increased the rate of dopamine synthesis and was ineffective in modifying the cyclic AMP levels in either normoxic or hypoxic CBs. 6. It is concluded that chemoreceptor cells are the intraglomic targets for almitrine. The mechanisms of action of almitrine on chemoreceptor cells are discussed.

Almitrine↗
PubMed1992

Characterization of cultured chemoreceptor cells dissociated from adult rabbit carotid body.

Short-term cell cultures were obtained from enzymatically dissociated carotid bodies from adult rabbits, and morphological and functional characterization of the cultured chemoreceptor cells were carried out. Under phase contrast, freshly isolated type I cells are round, bright, and 10-14 microns in diameter and exhibit strong fluorescence when stained with the glyoxylic acid technique. The content of endogenous dopamine in the cultures increased from 80 pmol/10(5) cells 2 h after plating the cells to 200 pmol/10(5) cells on the 3rd day, and the rate of synthesis and storage of [3H]dopamine from the precursor [3H]tyrosine increased from 1.7 pmol.10(5) cells-1.h-1 in 1-day cultures to 4 pmol.10(5) cells-1.h-1 on the 3rd day; the later values represent 80-85% of the expected values for the intact carotid body. After labeling with [3H]tyrosine, cultured chemoreceptor cells release [3H]dopamine when challenged by hypoxia, high external K+, or the protonophore dinitrophenol, the pattern of response being similar to that of the intact carotid body. When studied by whole cell clamp recording, individual chemoreceptor cells exhibit a marked variability in the properties of some ionic currents; the data, however, do not support the existence of distinct subpopulations of type I cells.

Animals↗
PubMed1991

Ionic mechanisms for the transduction of acidic stimuli in rabbit carotid body glomus cells.

1. The release of [3H]dopamine (DA) in response to inhibition of the Na+ pump or to intracellular acid load was studied in rabbit carotid bodies (CB) previously incubated with the precursor [3H]tyrosine. The ionic requirements of the release response and the involvement of specific ion transport systems were investigated. 2. Inhibition of the Na+ pump, by incubating the CB with ouabain or in K(+)-free medium, evokes a DA release response which requires the presence of Na+ and Ca2+ in the medium and is insensitive to nisoldipine. This suggests that the response is triggered by entry of external Ca2+ through Na(+)-Ca2+ exchange, a consequence of the increase in intracellular Na+ resulting from inhibition of the pump. 3. Incubation of the CB in medium equilibrated with 20% CO2 at pH 6.6, or in medium containing the protonophore dinitrophenol (DNP) or the weak acid propionate, elicits a DA release response which requires also the presence of Na+ and Ca2+ in the medium and is insensitive to dihydropyridines. 4. Ethylisopropylamiloride (EIPA), an inhibitor of the Na(+)-H+ exchanger, markedly decreases the release response elicited by DNP or propionate in bicarbonate-free medium, but has not any effect in bicarbonate-buffered medium. In the latter condition, the EIPA-insensitive release of DA is inhibited by reducing the HCO3- concentration in the medium to 2 mM or by removal of Cl-, suggesting that in bicarbonate-buffered medium a Na(+)-dependent HCO3(-)-Cl- exchanger is involved in the release response. 5. It is concluded that the release of DA by the chemoreceptor cells in response to acidic stimulation is triggered by entry of external Ca2+ through Na(+)-Ca2+ exchange. This exchange is promoted by the increase of intracellular Na+ that results from the operation of Na(+)-coupled H(+)-extruding mechanisms activated by the acid load.

Acids↗
PubMed1989

Effects of cyanide and uncouplers on chemoreceptor activity and ATP content of the cat carotid body.

In cat carotid bodies (c.b.'s) incubated in vitro with [3H]tyrosine to label the stores of catecholamines, it was found that CN promotes dose- and Ca2+-dependent release of [3H]dopamine (DA) from c.b. tissues in parallel to the increased electrical activity recorded from the carotid sinus nerve (c.s.n.). Two different uncouplers, dinitrophenol (DNP) and carbonyl-cyanide-m-chlorophenyl-hydrazone (CCCP), both activate also in a dose-dependent fashion, release of DA and electrical activity in the c.s.n. However, while cyanide (CN) (10(-4) M) applied during 5 min reduced the adenosine triphosphate (ATP) content of the c.b. by 45%, DNP (2.5 x 10(-4) M) and CCCP (10(-6) M) applied for the same period of time did not modify the ATP levels of the organ. At the above concentrations, the 3 agents increased about 8-fold the electrical activity recorded from the c.s.n. Thus, contrary to the postulates of the metabolic hypotheses, our findings indicate that the decrease in the ATP content in the c.b. is not a prerequisite for the activation of the chemoreceptors. We propose alternative mechanisms to explain the chemostimulant action of the metabolic poisons.

Action Potentials↗
PubMed1989

Metabolic activation of carotid body glomus cells by hypoxia.

The effects of low O2 on glucose consumption in the rabbit carotid body were studied using the in vitro 2-deoxyglucose technique. Metabolically active structures within the tissue were localized autoradiographically after freeze-drying and vacuum fixation/embedding of selected incubated tissue samples. In 100% O2-equilibrated media, the mean basal glucose consumption calculated from the rate of 2-[1,2-3H]deoxy-D-glucose phosphorylation and its specific activity in the incubation media was 61 nmol.g tissue-1.min-1 in the carotid body and 42 nmol.g tissue-1.min-1 in parallel experiments with nodose ganglia. Low PO2 (20% O2-equilibrated media in vitro) increased glucose consumption in the carotid body by 44% but did not alter glucose metabolism of nodose ganglia. Autoradiographic data showed that preneural type I parenchymal cells are the principal site of glucose consumption in carotid chemosensory tissue. The mechanisms responsible for the hypoxia-induced increase in glucose consumption by the type I cells are discussed in relation to sensory transduction by the carotid body chemoreceptors.

Animals↗
PubMed1988

Activation of the release of dopamine in the carotid body by veratridine. Evidence for the presence of voltage-dependent Na+ channels in type I cells.

Veratridine (50 microM), an agent known to activate voltage-dependent Na+ channels, induced a strong release of [3H]dopamine from the rabbit carotid body in vitro. The effect of veratridine was dependent on the presence of both Na+ and Ca2+ in the extracellular medium and was abolished by 1 microM tetrodotoxin. These results suggest that chemoreceptor type I cells have voltage-dependent Na+ channels, which could be involved in the depolarization of the cell membrane and activation of voltage-dependent Ca2+ channels.

Animals↗
PubMed1986

Effects of 2-deoxy-D-glucose on in vitro cat carotid body.

The process of chemosensory transduction in the arterial chemoreceptors is not well understood. According to the metabolic hypothesis of chemoreception, a decrease in arterial pO2 will produce a decrease in the adenosine triphosphate (ATP) content in the chemosensory type I cells, leading to release of a neurotransmitter and increased sensory neural activity. There is a paucity of direct experimental support for this hypothesis, and in the present work, we have tested the postulates of the metabolic hypothesis in an in vitro preparation of cat carotid body using 2-deoxy-D-glucose as an ATP-depleting agent. This preparation, when superfused with Tyrode containing 5 mM Na+-pyruvate instead of glucose, responds normally to hypoxia, low pH and acetylcholine, and maintains normal ATP levels. Under these conditions, 2-deoxy-D-glucose is a chemostimulant, i.e. electrical activity in the carotid sinus nerve is increased, with a threshold concentration of 0.25 mM and a maximum response at about 2-4 mM. These concentrations of 2-deoxyglucose evoke a dose-dependent release of [3H]dopamine (synthesized from [3H]tyrosine) from the carotid bodies which parallels the electrical activity. The 2-deoxy-D-glucose-evoked release and electrical activity is dependent on the presence of extracellular Ca2+. These same concentrations of 2-deoxy-D-glucose significantly reduce the ATP content of the carotid bodies. The events postulated by the metabolic hypothesis, i.e. decrease in ATP content, release of a putative neurotransmitter and activation of the sensory nerve endings are found to occur simultaneously. A possible cause-effect relationship between these three events is discussed.

Action Potentials↗
PubMed1986

Effects of high potassium on the release of [3H]dopamine from the cat carotid body in vitro.

Using an in vitro preparation of the cat carotid body, we have characterized the release of [3H]dopamine (DA) induced by high extracellular K+. Pulse superfusion (3 min) with high K+ Tyrode solution (20-80 mM) evoked a concentration-dependent release of [3H]DA from type I cells with a threshold at about 20 mM-extracellular K+ and a plateau at about 60 mM-extracellular K+. Equivalent low extracellular Na+ concentration ([Na+]o) solutions osmotically balanced with sucrose did not induce release. The high extracellular K+ concentration ([K+]o)-evoked release of [3H]DA by type I cells was dependent on the presence of Ca2+ in the superfusion media. On prolonged (10-14 min) superfusion with high K+ Tyrode solution, the [3H]DA release evoked by 60 mM-extracellular K+ was transient, while that evoked by 30 mM-extracellular K+ was sustained. In preparations superfused for 6 min with 60 mM-extracellular K+ and zero extracellular Ca2+ concentration ([Ca2+]o) Tyrode solution, reintroduction of Ca2+ did not elicit a secretory response. Ba2+ was a potent secretagogue of [3H]DA in preparations superfused with normal and zero [Ca2+]o Tyrode solution. Additionally, Ba2+ was capable of eliciting a secretory response from type I cells in preparations previously exposed (6 min) to 60 mM-extracellular K+, whether or not [Ca2+]o was present. With regards to stimulus-secretion coupling, our results indicate that high [K+]o probably depolarizes type I cells. This effect would, in turn, activate voltage-dependent Ca2+ channels, allowing the entrance of this ion to activate the neurosecretory response.

Action Potentials↗
PubMed1985

Correlation between adenosine triphosphate levels, dopamine release and electrical activity in the carotid body: support for the metabolic hypothesis of chemoreception.

An unsolved issue for the arterial chemoreceptors is the mechanism by which hypoxia and other natural stimuli lead to an increase of activity in the carotid sinus nerve. According to the 'metabolic hypothesis', the hypoxic activation of the carotid body (CB) is mediated by a decrease of the ATP levels in the type I cells, which then release a neurotransmitter capable of exciting the sensory nerve endings. Using an in vitro preparation of cat CB, we report that ATP levels in the CB do in fact decrease when the organs are exposed to moderate, short lasting hypoxia (5 min 20% O2). Additionally, we found that decreases in ATP levels induced by 2-deoxyglucose (2 mM) or sodium cyanide (0.1 mM) are closely correlated with dopamine release from type I cells and electrical activity in the carotid sinus nerve elicited by these agents. The possible cause-effect relationship of these events is discussed.

Adenosine Triphosphate↗