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J Alcayaga

Publications and source records attributed to J Alcayaga.

At least 19 recordsLinked to original sources

Dopamine modulates carotid nerve responses induced by acetylcholine on the cat petrosal ganglion in vitro.

We have recently reported that application of acetylcholine (ACh) or nicotine to the petrosal ganglion-the sensory ganglion of the glossopharyngeal nerve-elicits a burst of discharges in the carotid nerve branch, innervating the carotid body and sinus, but not in the glossopharyngeal branch, innervating the tongue and pharynx. Thus, the perikarya of sensory neurons for the carotid bifurcation exhibit selective cholinosensitivity. Since dopamine (DA) modulates carotid nerve chemosensory activity, we searched for the presence of DA sensitivity at the perikarya of these neurons in the cat petrosal ganglion superfused in vitro. Applications of DA in doses of up to 5 mg to the ganglion did not modify the rate of spontaneous discharges in the carotid nerve. However, if DA was applied 30 s before ACh injections, ACh-evoked reactions were modified: low doses of DA enhanced the subsequent responses to ACh, while high doses of DA depressed the responses to ACh. This depressant effect of DA on ACh responses was partially antagonized by adding spiroperone to the superfusate. Our results show that the response to ACh of petrosal ganglion neurons projecting through the carotid nerve is modulated by DA acting on D(2) receptors located in the somata of these neurons. Thus, dopaminergic modulation of cholinosensitivity could be shared also by the membranes of peripheral endings and perikarya of primary sensory neurons involved in arterial chemoreception.

Acetylcholine

Modulatory effect of nitric oxide on acetylcholine-induced activation of cat petrosal ganglion neurons in vitro.

The inhibitory effect of nitric oxide (NO) on carotid chemosensory responses to hypoxia has been attributed in part to an antidromic inhibition of chemoreceptor cells activity. However, NO may also modulate the activity of the primary sensory neurons because NO is produced in the soma of these neurons located in the petrosal ganglion. Since a population of petrosal neurons is selectively activated by acetylcholine (ACh), we studied the effects of NO-donor, sodium nitroprusside (SNP), and the NO-synthase inhibitor, Nomega-nitro-l-arginine methyl ester (l-NAME), on the responses evoked in the carotid sinus nerve (CSN) by ACh applied to the petrosal ganglion in vitro. ACh (1 microgram-1 mg) increased the frequency of action potentials recorded from the CSN in a dose-dependent manner. SNP (10-50 microM) reduced the sensibility and amplitude of the CSN response to ACh, although the maximal response appears less affected. The withdrawal of SNP from the superfusion medium increased the sensibility of the responses to ACh. l-NAME (1-2 mM) slightly increased the sensibility of the ACh-induced responses, effect that persisted after l-NAME withdrawal. These results suggest that NO may play a role as modulator in this autonomic primary sensory ganglion.

Acetylcholine

Sodium nitroprusside blocks the cat carotid chemosensory inhibition induced by dopamine, but not that by hyperoxia.

We studied the effects of the nitric oxide (NO) synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME), and the NO donor, sodium nitroprusside (SNP) on cat chemosensory responses to intravenous injections of NaCN (0.1-100 microg/kg) and dopamine (0. 1-20 microg/kg), and to hyperoxic ventilation (100% O2, 60-120 s). Cats were anesthetized with sodium pentobarbitone, paralyzed and artificially ventilated to prevent secondary ventilatory effects. The frequency of chemosensory discharges (fx) was recorded from one sectioned carotid sinus nerve. L-NAME (50 mg/kg i.v.) increased basal fx and slightly potentiated the responses to NaCN and dopamine. SNP (1-2 mg/kg i.v.) increased basal fx, but reduced the NaCN-induced increases of fx over baseline and the transient fx inhibitions induced by dopamine, but not those produced by hyperoxia. Present results indicate that besides the known inhibitory effect of NO on chemosensory responses to low PO2, NO also blocks the chemosensory response to dopamine, leaving hyperoxic responses largely unchanged.

Animals

Selective activation of carotid nerve fibers by acetylcholine applied to the cat petrosal ganglion in vitro.

The petrosal ganglion innervates carotid body chemoreceptors through the carotid (sinus) nerve. These primary sensory neurons are activated by transmitters released from receptor (glomus) cells, acetylcholine (ACh) having been proposed as one of the transmitters involved in this process. Since the perikarya of primary sensory neurons share several properties with peripheral sensory endings, we studied the electrical responses of the carotid nerve and glossopharyngeal branch to ACh locally applied to the cat petrosal ganglion superfused in vitro. Ganglionar applications of AChCl (1 microg-1 mg) generated bursts of action potentials conducted along the carotid nerve, while only a few spikes were exceptionally recorded from the glossopharyngeal branch in response to the largest doses. Carotid nerve responses to ACh were dose-dependent, the higher doses inducing transient desensitization. Application of nicotine to the petrosal ganglion also evoked dose-dependent excitatory responses in the carotid nerve. Responses to ACh were reversibly antagonized by adding hexamethonium to the superfusate, more intense and prolonged block of ACh responses being produced by mecamylamine. Ganglionar applications of gamma-amino butyric acid and serotonin, in doses of up to 5 mg, did not induce firing of action potentials in any of the branches of the glossopharyngeal nerve. Our results indicate that petrosal ganglion neurons projecting through the carotid nerve are selectively activated by ACh acting on nicotinic ACh receptors located in the somata of these neurons. Thus, cholinosensitivity would be shared by the membranes of peripheral endings and perikarya of primary sensory neurons involved in arterial chemoreception.

Acetylcholine

Effects of CO2-HCO3- on catecholamine efflux from cat carotid body.

Using a chronoamperometric technique with carbon-fiber microelectrodes and neural recordings, we simultaneously measured the effects of the following procedures on catecholamine efflux (delta CA) and frequency of chemosensory discharges (fx) from superfused cat carotid body: 1) the addition of CO2-HCO3- to Tyrode solution previously buffered with N-2-hydroxyethylpiperazine-N'-2-ethane-sulfonic acid, maintaining pH at 7.40; 2) hypercapnia (10% CO2, pH 7.10); 3) hypoxia (PO2 h approximately 40 Torr) with and without CO2-HCO3-; and 4) the impact of several boluses of dopamine (DA; 10-100 micrograms) on hypoxic and hypercapnic challenges. With CO2-HCO3-, hypoxia increased fx which preceded delta CA increases, whereas hypercapnia raised fx but did not consistently increase delta CA. Repeated stimuli induced similar fx increases, but attenuated delta CA. After DA, hypoxia produced larger delta CA, which preceded chemosensory responses. Without CO2-HCO3-, hypoxia produced a similar pattern of delta CA and fx responses. Switching to Tyrode solution with CO2-HCO3- at pH 7.40 raised fx but did not increase delta CA. With CO2-HCO3- and after DA, hypoxic-induced delta CAs were larger than in its absence. Results suggest that DA release is not essential for chemosensory excitation.

Animals

Cat carotid body chemosensory responses to non-hypoxic stimuli are inhibited by sodium nitroprusside in situ and in vitro.

We studied the effects of sodium nitroprusside, a nitric oxide donor, on the chemosensory responses to cyanide and nicotine in the cat carotid body. In situ, sodium nitroprusside infusion reduced the cyanide-evoked responses in a dose-dependent manner. In vitro, Tyrode containing nitroprusside reversibly reduced the cyanide- (by 59%) and nicotine-induced (by 45%) chemosensory responses. The present results suggest that chemosensory responses induced by cyanide and nicotine are reduced by increased nitric oxide content, similarly to the hypoxic chemosensory responses.

Animals

Time structure, temporal correlation and coherence of chemosensory impulses propagated through both carotid nerves in cats.

In spontaneously breathing, pentobarbitone anesthetized cats, we recorded simultaneously the impulses in the chemosensory fibers of both carotid (sinus) nerves, to analyze the correlations between the frequencies of chemosensory discharges (f chi) and their activation (¿df chi/dt¿a) and deactivation (¿df chi/dt¿d) rates. We studied the chemosensory responses to brief exposures to hypoxia (100% N2; 5-s and 10-s) and hyperoxia (100% O2; 30-s), and intravenous injections of excitatory (NaCN 0.2-100 micrograms/kg) and inhibitory (dopamine hydrochloride 0.02-20 micrograms/kg) chemoreceptor agents. Hypoxia increased f chi, with a high temporal correlation between frequency levels in both nerves. Prolonging hypoxic stimulation increased ¿df chi/dt¿d, with preservation of ¿df chi/dt¿a. Hyperoxic exposure produced highly correlated decreases in f chi in both nerves, but reduced correlation in df chi/dt. Increasing doses of NaCN produced analogous increments in f chi, df chi/dt and their correlations, the ¿df chi/dt¿a/¿df chi/dt¿d ratio remaining constant along all the experimental range, except in one animal in which the ratio increased in both nerves alike. Dopamine reduced f chi bilaterally, with chemosensory silencing being reached with doses of about 0.2-0.5 microgram/kg, the correlations between f chi's of both nerves remaining constant within the range analyzed. Maximal ¿df chi/dt¿d was not affected along the range of dopamine doses, except in one animal in which it increased in both nerves. It is concluded that both carotid nerves convey similar quantitative information to the brain stem. Thus, the carotid nerves constitute either cooperative inputs or redundant afferences contributing to a high safety factor.

Animals

Dissociation of hypoxia-induced chemosensory responses and catecholamine efflux in cat carotid body superfused in vitro.

1. To examine the correlation between chemosensory response and dopamine release induced by hypoxic stimulation, we studied carotid bodies excised from anaesthetized cats. 2. The carotid bodies with their carotid (sinus) nerves were superfused in vitro with modified Tyrode solution (pH 7.40, at 37.5 degrees C) equilibrated with 20 or 100% O2. The PO2 of the superfusing channel was monitored polarographically. The frequency of chemosensory discharges (fx) was recorded from the whole carotid nerve. Catecholamine (CA) efflux-mostly consisting of dopamine-was measured by high-speed chronoamperometry, through Nafion-coated carbon electrodes placed on the carotid body tissue. Chemosensory stimulation was induced by intrastream injections of NaCN, by superfusion with 100% N2-equilibrated saline (lowering PO2 to 25-40 Torr) or by flow interruption. 3. Low doses of NaCN increased fx, but had no measurable effect on CA efflux, while larger doses produced fast increases in fx, preceding delayed and prolonged increases in CA efflux. Repeated injections of NaCN, still increasing fx, gave reduced CA effluxes. 4. Switching to hypoxic superfusion for 6-8 min produced large and fast fx increases, but delayed and prolonged augmentations of CA efflux. 5. Administration of three to four boluses of dopamine (7-15 micrograms; augmenting CA concentration by up to 35 microM) initially decreased fx, after which hypoxic stimulation resulted in enhanced and faster CA effluxes, without changing the speed and intensity of chemosensory responses. 6. Flow interruptions induced fast increases in fx and delayed increases in CA efflux. Repeated flow interruptions produced similar increases in fx but progressively attenuated CA effluxes. 7. Our results suggest that CA efflux is not essential for hypoxia-induced chemosensory excitation in the cat carotid body. They also suggest the presence of two pools of releasable CAs in the carotid body, one of slow turnover and release, and another of recently incorporated dopamine and fast release, both pools being rapidly depleted by repeated stimulation of the carotid body.

Animals

The petrosal ganglion of the adult cat: neuronal count, sectional area, and their respective distributions.

The petrosal ganglion contains most of the perikarya of sensory neurons of the glossopharyngeal nerve. We studied the number and size of neuronal somata in 4 petrosal ganglia from adult cats. Ganglia were serially sectioned in length at 8 microns, sections drawn through a projection microscope, and those neuronal profiles presenting nuclei and nucleoli on each section were counted and their areas measured. The number of neurons ranged from 2311 to 3429 (2908 +/- 271; mean +/- SEM). Neurons were symmetrically distributed around the longitudinal axes of most ganglia, with a skewed distribution in only one ganglion. The sectional area of most neurons (> 98%) ranged between 250 and 1725 microns 2, with median values of 667-963 microns 2. Area distributions were significantly different, but differences never exceeded 8.2% in related area bins. The ganglion presenting a skewed count distribution and the highest median area departed from the rest, with differences surpassing 25%. We conclude that the neuronal population of the petrosal ganglion of the cat is regular both with respect to the number and the size of its constituents, with departures from this pattern probably reflecting individual variations.

Adult

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% O2, thermally dependent increases in frequency were observed, with significant differences between all nearby thermal stages separated by 0.5 degrees C steps between 36.0 and 38.5 degrees 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% O2 and no significant differences in mean ranks were recorded between 36 and 37 degrees C, as between 38 and 39 degrees C, but frequencies at 36-37 degrees C were significantly higher than those at 38-39 degrees C. Brief rises in chemosensory discharges were evoked by injections of NaCN applied to carotid bodies superfused with saline equilibrated with 100% O2. The least effective dose was lower at 40 degrees C than at 37.5 degrees or 35.0 degrees 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.

Animals

The chick chorioallantoic membrane promotes survival of co-transplanted rat carotid bodies and nodose ganglia.

Carotid bodies and nodose ganglia, removed from adult rats, were co-implanted onto the chorioallantois of 6- to 12-day chick embryos. Implants were rapidly vascularized and incorporated into the chorioallantoic membrane, where they survived and grew for up to 12 days. The morphological characteristics of grafted tissues were largely preserved. Regenerating axons from nodose neurons invaded the carotid body and contacted some glomus cells through morphologically immature synapses. Thus, the chick chorioallantoic membrane may be a useful substrate to study carotid chemoreceptor-sensory neuron interactions.

Allantois

Electrophysiological evidence for the reconstitution of chemosensory units in co-cultures of carotid body and nodose ganglion neurons.

The electrophysiological characteristics of nodose ganglion sensory neurons, cultured alone or co-cultured with carotid body tissue, were compared. Some properties of the neurons and their response to acid (a carotid body 'natural' stimulus) changed in the presence of this tissue. (a) The evoked action potential after-hyperpolarization was smaller and longer whereas spike amplitude and duration, and the passive membrane properties remained unaltered. (b) Spontaneously occurring action potentials happened more frequently (16% vs 3%). (c) Acid solutions induced appreciable depolarization, an increased discharge, or both, only in a population of co-cultured neurons. These changes probably arose because of synaptic and/or trophic interactions between neurons and glomus cells.

Action Potentials

Fast activity and oscillatory potential of carp retina in the frequency domain.

There are two kinds of fast activity in the ERG: fast retinal potentials (FRP), an irregular series of spiky wavelets and oscillatory potentials (OP), a rhythmic sequence of events. Corneal ERG from nine intact young carps, evoked by extended pulses of diffuse white light under mesopic adaptation, displayed two different groups of wavelets related to ON and OFF, respectively. Stimulation and recording conditions were established to permit separate Fourier analysis of both groups of wavelets. Power distributions of normalized ON spectra showed both a wide dispersion and a high inter-subject variability. All normalized OFF spectra showed, instead, components within a narrow band from 52 to 56 Hz, most of them maximum relative power peaks. It is concluded that FRP originating in highly labile sources dominate ON fast activity, while the predominant OFF fast activity are OP originating in a stable discrete source.

Animals

CONFREG: a BASIC program for calculating and plotting confidence regions based on correlational analyses.

Many observations encountered in biological and medical research are randomly distributed in bivariate scales, and thus not susceptible to simple regression analyses. Since such data are depicted by ellipses in scatter diagrams, a computer program to calculate the confidence regions for the means or the total data of bivariate samples was written in BASIC for correlational analyses. The program, based on the principal axes algorithm, plots the calculated confidence regions as an elliptic area, using the fitted equations for its major and minor axes. The program displays the sample parameters required to perform comparisons between different groups of experimental conditions.

Animals