Search PubMed⌕ Search

Biomedical subjects

J Alcayaga

Publications and source records attributed to J Alcayaga.

At least 19 recordsLinked to original sources
PubMed2000

Acetylcholine sensitivity in sensory neurons dissociated from the cat petrosal ganglion.

The petrosal ganglia contain the somata of the sensory fibers of the glossopharyngeal nerves, innervating structures of the tongue, pharynx, carotid sinus and carotid body. Petrosal ganglia were excised from adult cats and their neurons were dissociated and kept in tissue culture for 7-12 days. Intracellular recordings were obtained through conventional microelectrodes. In response to depolarizing pulses, most cells (41/60) presented a 'hump' in the falling phase of their action potentials (H-type), while the remaining neurons lack such hump (F-type). The two types of cells had no differences in resting membrane potential or action potential amplitude. Acetylcholine (ACh) applied locally elicited responses in nearly two thirds of both H-type and F-type neurons tested. Most H-type neurons (17/19) responded with a slow long lasting depolarization, while the remaining (2) did so by generating spikes. In contrast, half of F-type neurons (6/12) responded with one or more spikes and the other half only with a slow depolarization. These results indicate that ACh receptors are present in the soma of many petrosal ganglion neurons subjected to tissue culture, thus supporting the idea that - under normal conditions - their peripheral sensory processes may be excited by ACh.

Acetylcholine↗
PubMed2000

Lack of correlation between cholinergic-induced changes in chemosensory activity and dopamine release from the cat carotid body in vitro.

We studied the effects of nicotine, acetylcholine (ACh) and dopamine (DA) on the frequency of chemosensory discharges (f(x)) and catecholamine (CA) efflux in the cat carotid body superfused in vitro. CA efflux was measured by changes in CA concentration (DeltaCA) determined by chronoamperometry with nafionated carbon-fiber microelectrodes inserted in the carotid body, while f(x) was recorded simultaneously from the carotid (sinus) nerve. Nicotine (10-20 microg) and ACh (>100 microg) increased f(x) in all carotid bodies (n=16), but produced a delayed DeltaCA ( approximately 0.65 microM) in only half of them. Eserine potentiated ACh-evoked increases in f(x) and CA effluxes. Nicotine and ACh-induced DeltaCA were rapidly reduced upon repeated administration. While f(x) increases evoked by low doses of nicotine or ACh were reduced or abolished by prior administration of exogenous DA (>100 microg), CA effluxes were enhanced and hastened. Thus, cholinergic-induced changes in f(x) are dissociated from CA efflux.

Acetylcholine↗
PubMed2000

Adenosine triphosphate-induced peripheral nerve discharges generated from the cat petrosal ganglion in vitro.

Since nucleotides have been postulated as transmitters between glomus cells and chemosensory nerve endings in the carotid body, we studied the effects of their application to the petrosal ganglion, where the perikarya of carotid (sinus) nerve are located. Cat petrosal ganglia were superfused in vitro, while electrical activities of their peripheral processes (carotid nerve and glossopharyngeal branch) were recorded simultaneously. Adenosine triphosphate (ATP) evoked dose-dependent bursts of impulses in carotid nerve, while those in glossopharyngeal branch were less intense and consistent. Adenosine monophosphate was less effective than ATP. ATP-induced carotid nerve responses presented no temporal desensitization and persisted after applying P(2Y) receptor blocker Reactive Blue 2 to the ganglion. The results indicate that ATP has an excitatory effect on the perikarya of the population of petrosal ganglion neurons projecting peripherally through the carotid nerve.

Action Potentials↗
PubMed1999

Responses to hypoxia of petrosal ganglia in vitro.

NaCN is a classical stimulus used to elicit discharges from carotid body chemoreceptors. The effect is assumed to be mediated by glomus (type I) cells, which release an excitatory transmitter for the excitation of carotid nerve endings. Since the sensory perikarya of the glossopharyngeal nerve (from which the carotid nerve branches) are located in the petrosal ganglion, we tested whether application of this drug to the petrosal ganglion superfused in vitro elicits antidromic discharges in the carotid nerve. NaCN did indeed cause an intense and prolonged burst of nerve impulses in the carotid nerve, while provoking a less intense and much briefer burst of discharges in the glossopharyngeal branch. Carotid nerve responses to NaCN were reduced and shortened by prior or following application of dopamine to the ganglion. Sodium azide applied to the petrosal ganglion evoked a less intense and much briefer burst of impulses in the carotid nerve. Ganglionar application of 2,4-dinitrophenol did not induce discharges in the carotid nerve. Switching the superfusion of the ganglion from a normoxic to a hypoxic solution did not evoke discharges in the carotid nerve. Therefore, the perikarya of carotid nerve neurons are sensitive to NaCN, but are not excited by reducing the pO(2) of the superfusing solution.

2,4-Dinitrophenol↗
PubMed1999

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↗
PubMed1999

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↗
PubMed1998

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↗
PubMed1998

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↗
PubMed1998

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↗
PubMed1997

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↗
PubMed1997

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↗
PubMed1996

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↗