Calcium concentration of salivary glands and saliva after chemical or surgical sympathectomy

Calcium concentration of the submaxillary (SM) gland of adult rat was increased to 2-3 times control levels 24 h after a single high dose of reserpine (RES) (5 mg/kg body weight), 6-hydroxydopamine (6-OHDA) (20 mg/kg body weight), or surgical removal of a superior cervical ganglion (Sx). The increase could also be induced within 24 h after a single injection of lower doses of RES (0.05, 0.5 mg/kg body weight). Increase in calcium concentration in the parotid (PA) was not found under any of the above conditions, but did occur transiently at times within 24 h. The increase in glandular calcium concentration was found to be temporally related to depletion of norepinephrine (NE). The time frame for initiation of calcium changes was not the same for chemical and surgical sympathectomy (Sx). Increase in calcium concentration occurred after onset of sympathectomy-induced degeneration secretion; the onset varied with the kind of sympathectomy, and was seen 1 h after drug administration with either RES or 6-OHDA, but not until about 13 h after surgical Sx. Similarly, NE was markedly depleted, but later with surgical than with chemical Sx. Furthermore, with both kinds of sympathectomy, depletion of NE occurred earlier with PA than with SM, since normal basal levels of NE of SM are 3 times as great (3400 ng/g wet weight), compared with PA levels (1300 ng/g wet weight). The fact that NED was the neurotransmitter eliciting the degeneration secretion was confirmed physiologically by examination of the composition of this secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Beta-adrenergic receptors and cAMP levels of rat parotid and submandibular glands during chronic isoproterenol treatment

The number of cell surface beta-adrenergic receptors and the level of cyclic AMP of the parotid and the submandibular gland were examined in rats treated for up to 10 days with twice daily injections of the beta-adrenergic agonist, isoproterenol. Receptor densities of 125 +/- 8.7 fmol/mg membrane protein for the parotid and 60.1 +/- 5.6 fmol/mg for the submandibular glands were found with [3H]dihydroalprenolol (beta-adrenergic receptor antagonist) binding of glands from control rats. No change from levels of controls was found in the number of beta-receptors of the submandibular gland with chronic isoproterenol stimulation; the parotid glands, on the other hand, showed a 22% decrease in dihydroalprenolol binding from the 4th until the 8th day of treatment. By day 10 of isoproterenol treatment the parotid gland demonstrated a shift from a population consisting of primarily beta-adrenergic receptors to one consisting of equal numbers of beta 1- and beta 2-adrenoceptors. The basal level of cAMP present in cell lysates remained unchanged in the isoproterenol-treated submandibular gland while the parotid gland showed a 30-40% decrease. Control and isoproterenol-treated animals demonstrated the same time course of cAMP accumulation after a single challenge with isoproterenol.     

Evidence for a mutual interaction between noradrenergic and serotonergic agonists in stimulation of ACTH and corticosterone secretion in the rat

We investigated the mutual interactions between hypothalamic norepinephrine (NE) and serotonin (5-HT) in mediating the ACTH and corticosterone responses to direct stimulation of the paraventricular nucleus (PVN) with adrenergic and serotonergic agonists. The hormone responses to the intrahypothalamic injection of the alpha1-adrenergic agonist phenylephrine (20 nmol/2 microl) were significantly reduced by prior depletion of hypothalamic 5-HT with intra-PVN injection of the serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT), but not after depletion of hypothalamic NE by intra-PVN injection of the noradrenergic neurotoxin 6-hydroxydopamine (6-OHDA). The ACTH and corticosterone responses to intrahypothalamic injection of the 5-HT(1A) receptor agonist 8-OH-DPAT (20 n mol/2 microl) were significantly reduced by depletion of hypothalamic NE with 6-OHDA, but not after depletion of hypothalamic 5-HT with 5,7-DHT. These mutual interactions between the NE and 5-HT neuronal systems, which innervate the PVN, may explain previous findings of equivalent reductions in the hypothalamic-pituitary-adrenal axis responses to neural stimulation after neurotoxic lesioning of either the NE or 5-HT systems.     

Cyclosporin A: effects on the secretory process and noradrenergic activity in the submandibular gland of the rat.

OBJECTIVES: The aim of the present work was to study the effect of long-term cyclosporine (CSA) administration on norepinephrine (NE) metabolism and adrenergic-evoked secretion in the rat submandibular gland (SMG). METHODS: Dose-response curves to adrenergic agonists (methoxamine, isoproterenol, NE) were performed in control and CSA (10 and 30 mg/kg every 2 days for 1 month)-treated rats after SMG duct cannulation. In SMG tissue neuronal NE uptake, release, synthesis and endogenous content were determined. In addition phosphoinositide intracellular signaling was also investigated. RESULTS: CSA administration caused an increase in salivary secretion evoked by methoxamine (alpha-adrenergic agonist) and NE but failed to modify salivation evoked by beta-adrenergic stimulation (isoproterenol). Long-term CSA administration decreased NE release and synthesis whereas it enhanced the amine uptake and phosphoinositide hydrolysis in the SMG. CONCLUSIONS: The administration of CSA for 30 days induced salivary gland sensitization likely mediated by diminished adrenergic input. Present results suggest that the decreased sympathetic activity evoked by long-term CSA administration in the rat SMG may lead to sensitization of the gland supported by increased phosphoinositide hydrolysis and enhanced adrenergic-evoked salivation.     

Effects of adrenergic agonists on electrolyte transport in perfused salivary duct of rat

The Na and Cl absorption and K secretion that occur in the main duct of rat submandibular gland are affected by adrenergic actions. The specific effects of stimulation of alpha-, beta 1- and beta 2-adrenergic receptors on net transepithelial fluxes of Na, K and Cl were investigated in microperfused main excretory duct of rat submandibular gland. Administration of methoxamine (2.5 micrograms . kg-1 . min-1, i.v.) resulted in marked decreases in net efflux (or absorption) of Na (38%) from the duct into the interstitial fluid and net influx (or secretion) of K (20%) from interstitial fluid into the ductal lumen without any effect on net efflux or reabsorption of Cl. A higher dose of methoxamine (5 micrograms . kg-1 . min-1, i.v.) produced further inhibition of net fluxes of Na and K without affecting net flux of Cl. Dobutamine (50 micrograms . kg-1 . min-1, i.v.) enhanced net effluxes of Na (40%) and Cl (300%) from the lumen but did not alter net K influx into the lumen. When the dosage of dobutamine was increased to 200 micrograms . kg-1 . min-1, an inhibition of net influx of K (28%) into ductal fluid was observed in addition to enhanced net fluxes of Na and Cl from the lumen. Administration of terbutaline (15 and 30 micrograms . kg-1 . min-1, i.v.) decreased net influx of K (30-40%) and increased net efflux of Cl (280%) without affecting net efflux of Na.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of cyclocytidine and sympathetic nerve induced changes in norepinephrine and adrenoceptors in salivary glands.

Norepinephrine (NE) concentration of parotid and submandibular glands of young rats was reduced 51% and 39%, respectively at 1 h, and 60% and 47% at 2 h after i.p. administration of a single dose (500 mg/kg body weight) of the anti-tumor agent, cyclocytidine (CC). For adult rats, the reductions were 44% and 46%, respectively, at 1 h and 54% and 49% at 2 h. This decrease from controls was generally similar to the decrease induced following 1 and 2 h of electrical stimulation (square wave pulses of 4 V, 5 ms duration, and frequency of 16 Hz) of the sympathetic innervation to these glands (young rats, 59% and 58% at 1 h; 66% and 63% at 2 h; for adult rats, 51% and 55% at 1 h and 69% and 53% at 2 h for parotid and submandibular, respectively). The changes in density of beta-adrenoceptors induced by direct nerve stimulation also corresponded to the changes induced by CC (CC induced a decrease in parotid of 12%, compared with a decrease of 11% with electrical stimulation; a 15% and 18% reduction in number of beta-adrenoceptors of submandibular gland was found at 1 h after CC and electrical stimulation, respectively). Compelling evidence for the mechanism of CC action was thus established, showing that CC mimics effects of sympathetic nerve stimulation (inducing reduction in NE concentration and transient change in beta-adrenoceptor density) by causing release of NE from sympathetic nerve endings.     

Norepinephrine-evoked calcium transients in cultured cerebral type 1 astroglia

We have used the CA++ indicator dye fura-2 AM and computerized imaging systems to investigate adrenergic regulation of intracellular calcium in cultured cerebral type 1 astroglia. We have found that norepinephrine (NE) and other adrenergic agonists stimulate increases in intracellular calcium in over 80% of type 1 astroglia tested. A wide range in effective NE concentrations was seen. With sufficient agonist concentrations the calcium response was biphasic, exhibiting an initial sharp peak followed by a sustained calcium elevation. This secondary component was sensitive to reductions in extracellular calcium concentrations and dependent on the continued presence of agonist. Pharmacological studies indicated that astroglial calcium responses were mediated by alpha 1- and alpha 2-adrenergic receptors. At times these two receptor subtypes appeared to underlie calcium responses by the same cells, whereas other cells only responded to stimulation of one or the other subtypes of alpha-adrenergic receptor. Finally, we have also observed spontaneous and agonist-evoked oscillations in astroglial calcium levels. The major findings of these studies indicate that 1) astroglial cells respond to alpha-adrenergic receptor stimulation with increased intracellular calcium, 2) these responses can be mediated by alpha 1-and/or alpha 2-adrenergic receptors, and 3) subpopulations of cerebral type 1 astroglia exist with respect to alpha-adrenergic receptor expression.     

Studies of muscarinic receptor subtypes in salivary gland function in anaesthetized rats

The in vivo study aimed to examine whether muscarinic receptor subtypes other than muscarinic M3 receptors exert exocrine functional roles in the rat salivary glands. The effects of pirenzepine, methoctramine and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) were examined on secretion from the major salivary glands evoked by acetylcholine (0.001-10 micromol kg(-1) i.v.) in pentobarbitone-anaesthetized rats. Observations were occasionally made on glandular blood flow. 4-DAMP (0.1-100 nmol kg(-1) i.v.) markedly and equipotently inhibited the acetylcholine-evoked fluid responses in all glands. Pirenzepine (0.1 micromol kg(-1) i.v.-10 mmol kg(-1) i.v.) showed significantly lower inhibitory potency than 4-DAMP, most conspicuously in the parotid, while methoctramine (0.1 micromol kg(-1) i.v.-10 mmol kg(-1) i.v.) exerted an even lesser inhibitory effect. Also against acetylcholine-evoked blood flow increases, 4-DAMP showed a conspicuous potency. At 1 and 10 micromol kg(-1) i.v. of pirenzepine, the antagonist reduced the protein concentration in the submandibular saliva, but not in the parotid saliva. While 4-DAMP (1 and 10 nmol kg(-1) i.v.) significantly inhibited acetylcholine-evoked protein secretory responses in the submandibular glands, methoctramine (below 10 micromol kg(-1) i.v.) affected the responses in neither gland. The reduction of the protein concentration in submandibular saliva caused by 4-DAMP and pirenzepine was inhibited by N(omega)-nitro-L-arginine methyl ester (L-NAME; 30 mg kg(-1) i.p.), while L-NAME had no or only minute effects on the parotid protein secretion. Thus, in addition to muscarinic M3 receptors, other muscarinic receptors contribute to in vivo functional responses in rat submandibular and sublingual glands. While these other receptors are muscarinic M1 receptors in the sublingual gland, they may be a different subtype, possibly muscarinic M5 receptors, in the submandibular gland. However, muscarinic M1 receptors may induce indirect effects via nitric oxide in the submandibular gland.     

beta2-adrenergic receptor activation and genetic polymorphisms in autism: data from dizygotic twins

Gestational and genetic factors can contribute to autism during infancy and early childhood through their effects on fetal brain development. Previous twin studies have shown strong genetic components for the development of autism, a disorder that can have multiple causes. We investigated the effects of prenatal overstimulation of the beta2-adrenergic receptor in dizygotic twins who were exposed to terbutaline, a selective beta2-adrenergic receptor agonist used to treat premature labor, as a gestational factor. As a possible genetic mechanism, we studied two beta2-adrenergic receptor polymorphisms in twins from whom DNA was available: glycine substitution at codon 16 (16G) and glutamic acid substitution at codon 27 (27E), which show diminished desensitization in vivo compared with the wild-type receptor. Continuous terbutaline exposure for 2 weeks or longer was associated with increased concordance for autism spectrum disorders in dizygotic twins (relative risk = 2.0), with a further increase in the risk for male twins with no other affected siblings (relative risk = 4.4). A significant association was found between the presence of 16G and 27E polymorphisms in autistic patients compared with population controls (P = .006). Prenatal overstimulation of the beta2-adrenergic receptor by terbutaline or by increased signaling of genetic polymorphisms of the beta2-adrenergic receptor that have diminished desensitization can affect cellular responses and developmental programs in the fetal brain, leading to autism.     

Cytokine production by naive and primary effector CD4+ T cells exposed to norepinephrine

We recently showed that clones of Th1 cells, but not Th2 cells, expressed a functional beta-2-adrenergic receptor (beta2AR) and that either norepinephrine or the beta2AR agonist terbutaline stimulated this receptor to modulate the level of Th1 cytokines produced. In the present study, we show that norepinephrine and terbutaline stimulate the beta2AR to decrease the level of IL-2 produced by freshly isolated murine splenic naive CD4+ T cells from either Balb/C or DO11.10 transgenic mice and activated polyclonally with anti-CD3 and anti-CD28 mAbs. In contrast, the level of cytokines produced by primary effector Th1 and Th2 cells were unaffected when norepinephrine, terbutaline, or cAMP analogs were added at the time of restimulation. These results suggest that a diversity exists among CD4+ T-cell subsets with respect to the level of adrenergic receptor expression, responsiveness to cAMP, stage of cell differentiation, or a combination of the above.     

Platelet-mediated vascular permeability in the rat: a predominant role for 5-hydroxytryptamine

The intradermal injection in rat skin of washed, thrombin-activated platelets produces an increase in vascular permeability, the intensity of which increments with the platelet concentration. Pretreatment of the recipient animals with serotonergic antagonists, including the specific 5-HT2 receptor blocker ketanserin, potently inhibits the platelet-mediated and the 5-HT-induced vascular defect. Amine depletion of platelets or skin tissues with reserpine reduces the response to platelets. Platelet prostanoid and lipoxygenase derivatives play no major role in the vascular response to platelet. The permeability increase induced by exogenous 5-HT and by activated platelets is reduced by alpha 1-adrenergic stimulation with noradrenaline or phenylephrine and by beta 2-stimulation with terbutaline or isoprenaline, and is potentiated by adenosine; this points to a modulation of permeability by blood flow changes and to a direct beta-adrenergic effect at the endothelial cell membrane. This study demonstrates a predominant role for 5-HT in the platelet-mediated vascular permeability increase in a sensitive species like the rat.     

Acute amitriptyline effects on parasympathetic-evoked rat saliva

The present study was undertaken to investigate the acute effects of amitriptyline on salivary secretion evoked by electrical stimulation of the parasympathetic innervations of rat salivary glands. Single intravenous injections of amitriptyline (0.1-1 mg/kg) caused a dose-related decrease in flow and Na concentration of saliva from both parotid and submandibular glands. However, the only effect on K concentration was a slight increase when the salivary flow was almost completely inhibited. Amitriptyline increased the Ca concentration of nerve-evoked submandibular saliva, but had no effect on the Ca concentration of similarly evoked parotid saliva. However, amitriptyline (0.5 and 1 mg/kg) increased the protein concentration of both kinds of saliva. Amylase activity of parotid saliva was also moderately increased by amitriptyline. These effects were similar to those observed with atropine, a known cholinergic receptor antagonist. These results suggest that amitriptyline, like atropine, reduces parasympathetic-evoked salivary secretion by blocking cholinergic receptors.     

Regions in the brainstem and frontal cortex where electrical stimulation elicits parotid and submandibular saliva secretion in sheep

Acute experiments were conducted in sheep anaesthetized with sodium pentobarbital to identify regions in the brain where electrical stimulation would elicit secretion by the parotid and submandibular salivary glands. This was a prerequisite for single unit studies in the future. In the brainstem, parotid and submandibular secretions were evoked on average between 7 and 16 mm rostral to obex and from 1 to 11 mm lateral to midline, with the parotid gland being active on the caudal and the submandibular on the rostral sides of this region. Overlap of the two sites was common. The combined region was either between cranial nerves VII and IX or adjacent to either one of them. Stimulating the caudal edge of the parotid sites evoked relatively high parotid secretion rates compared with other areas whereas secretions were uniform throughout the submandibular sites. The combined sites were from 2 to 5 mm deep, the dorsal edge being 1 mm below the floor of the fourth ventricle near midline and 6 mm below it at the lateral extremes. From a dorsal perspective, their orientation was essentially in the lateral plane except that the submandibular site angled slightly rostrally from midline. Profuse parotid secretion was also consistently evoked by stimulating the frontal cortex 15-20 mm from midline and 0-15 mm under the apex. Weak submandibular responses were observed in about half of the sheep. There were no effects on either gland of stimulating the olfactory bulbs. This is the first report of the regions in the brain which increase parotid and submandibular saliva secretion in ruminants. The three-dimensional representation of both sites in the brainstem of individual animals is more precise than the composite representations published to date for other species.     

Receptor subtype and dose dependence of dexmedetomidine-induced accumulation of [14C]glutamine in astrocytes suggests glial involvement in its hypnotic-sedative and anesthetic-sparing effects

Dexmedetomidine, a selective alpha(2)-adrenergic agonist, increases accumulation of [14C]glutamine and its labeled metabolites in primary cultures of mouse astrocytes. The concentration dependence is biphasic and identical to that previously described for dexmedetomidine's effect on free cytosolic calcium concentration ([Ca(2+)](i)) in astrocytes, and both effects are exerted on the alpha(2A) subtype of the alpha(2) receptor, suggesting a Ca(2+)-mediated effect. The concentration corresponding to the most potent effect is similar to that with which dexmedetomidine exerts its anesthetic-sparing activity in vivo, and the second peak corresponds to its hypnotic-sedative effect. It is suggested that both effects may be caused by decreased glutamatergic neurotransmission, secondary to reduced availability of glutamine as a glutamate precursor in glutamatergic neurons.     

Beta-adrenoceptor modulation of transiently overexpressed alpha 2-adrenoceptors in brain and peripheral tissues: cellular mechanisms underlying the developmental toxicity of terbutaline

Terbutaline, a selective beta(2)-adrenoceptor (beta(2)AR) agonist, is widely used as a tocolytic to arrest preterm labor but recent studies indicate that excessive betaAR stimulation can alter the expression and function of other neurotransmitter receptors that are essential to fetal/neonatal development. In many immature tissues, alpha(2)-adrenergic receptors (alpha(2)ARs) are overexpressed and the receptors are thought to play a role in cell proliferation and architectural assembly. We evaluated whether betaAR agonists perturb the expression of alpha(2)ARs in central and peripheral tissues during various developmental stages in the fetal and neonatal rat. In peripheral tissues (heart, liver, kidney) administration of terbutaline (10mg/kg s.c. for 4 days) elicited decrements in alpha(2)AR expression only during a critical developmental window that differed for each tissue; terbutaline was more effective than isoproterenol, a mixed beta(1)/beta(2) agonist. Neonatal destruction of sympathetic nerves with 6-hydroxydopamine (6-OHDA) had a biphasic effect, initially reducing alpha(2)ARs but subsequently elevating receptor expression. In contrast to the effects in the periphery, terbutaline administration promoted alpha(2)AR expression in neonatal brain regions with effects preferential to males. As the rat is an altricial species, these results during late gestation and the early neonatal period indicate that betaAR input modulates alpha(2)AR expression during developmental stages in which betaAR tocolytics are likely to be used. Disruption of alpha(2)AR expression and function may therefore contribute to adverse effects that have been noted in the offspring of pregnant women treated with terbutaline.     

Beta 2-adrenergic receptor agonist induces IL-18 production without IL-12 production

Endogenous catecholamine, epinephrine and norepinephrine, and isoproterenol concentration-dependently induced the production of interleukin (IL)-18, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma, and inhibited that of IL-10 in human peripheral blood mononuclear cells (PBMC). All responses by these stimulations were antagonized by the selective beta 2-adrenergic receptor (AR) antagonist, butoxamine, but not by alpha 1-, alpha 2- and beta 1-AR antagonists. The selective beta 2-AR agonists, salbutamol and terbutaline, induced a similar pattern of cytokine production, indicating that the effect of these AR agonists on cytokine production was through beta 2-AR stimulation. Anti-IL-18 Ab or caspase-1 inhibitor prevented all increase/decrease effects, suggesting that IL-18 might affect the production of all other cytokines. While endogenous IL-18 produced by salbutamol and terbutaline reached a sufficient concentration to induce IL-12 production, these beta 2-AR agonists did not induce the production of IL-12 at all. Epinephrine/norepinephrine/isoproterenol/beta 2-AR agonists increased the production of IL-18 in monocytes, but had no effect on IL-12, TNF-alpha, IFN-gamma and IL-10 production. The lack of beta 2-AR-induced effect on IL-12 production was due to a beta 2-AR-induced inhibition of an IL-18-elicited upregulation of both CD40 and CD40 ligand (CD40L/CD154) expressions on monocytes. The sympathetic innervating lymphoid organs may be under the control of beta2-AR stimulation, maintaining the basal cytokine environment in the tissues.     

Effects of noradrenergic agonists and antagonists on growth hormone secretion under gamma-hydroxybutyrate narco-analgesia in the rat

(1) The administration of gamma-hydroxybutyrate (GHB) has no effect on the hypothalamic concentration of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) in the rat. (2) The injection of GHB induces a consistent growth hormone (immunoreactive GH) secretory episode, followed by basal levels for several hr. (3) During this period, the infusion of clonidine, an α-adrenergic agonist, elicits a dramatic GH increment, whereas basal levels are not modified by isoproterenol, a β-adrenergic agonist. (4) In animals pretreated with α- and β-adrenergic agonists, clonidine enhances the GH peak induced by GBH, which is clearly inhibited by isoproterenol. The effects of α- and β-adrenergic antagonists were also investigated in this study. (5) The data reported here could support the hypothesis that α-adrenergic receptors mediate the secretion of the GH-releasing hormone and that β-adrenergic receptors are involved in the release of somatostatin.     

Beta-adrenoceptors in the rat parotid gland enlarged by salivariectomy and bulk diet. Effects of denervation

The numbers of beta-adrenergic receptors and level of cyclic AMP (cAMP) of the parotid gland of adult female rats were determined 4 weeks after introduction of a regimen that induced a 2-fold increase in gland weight. This regimen consisted of ablation of the submandibular-sublingual glands and substitution of the normal chow diet with a bulk diet consisting of 50% inert cellulose and 50% ground solid chow. There was a 2.4-fold increase in number (density) of beta-adrenoceptors in the enlarged parotid gland when comparison was made with parotid glands of control rats. The beta-adrenoceptor present in the enlarged and normal glands was of the beta 1 subtype. Removal of either autonomic pathway at the time of partial salivariectomy and dietary substitution was followed by a small reduction in number of beta-adrenoceptors (4-9% with either denervation), but when both nerves were removed the reduction was 25%; in magnitude, these changes were generally similar to those observed with denervated parotid glands of chow-fed rats. The norepinephrine concentration of the enlarged gland was much less than that of normal glands (reduced 38%); sympathectomy of normal or enlarged parotid glands resulted in a marked lowering of norepinephrine concentrations (to 1-5% of control levels); parasympathectomy had no effect on norepinephrine concentration of enlarged parotid glands but caused a decrease in that of the parotid of normal size. Apparently, the number of beta-adrenoceptors depends on the degree of activity of both the parasympathetic and sympathetic nerves to the parotid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Excitatory amino acid regulation of intracellular Ca2+ in isolated catfish cone horizontal cells measured under voltage- and concentration-clamp conditions.

[Ca2+]i was measured using fura-2-loaded isolated catfish horizontal cells in the presence of L-glutamate and the glutamate analogs kainate (KA), quisqualate (QA), and NMDA. Caffeine was used to release Ca2+ from intracellular stores. Cell membrane potential was controlled with a voltage clamp to prevent activation of voltage-dependent Ca2+ channels in the presence of agonist. All excitatory amino acid agonists produced a rapid and sustained rise in [Ca2+]i with the order of potency being QA greater than Glu greater than KA greater than NMDA. The agonist-induced [Ca2+]i increase was blocked in reduced [Ca2+]o and by 6-cyano-7-nitroquinoxaline-2,3-dione and 2-amino-5-phosphonopentanoate, which are specific blockers for QA/KA and NMDA receptors, respectively. The metabotropic receptor agonist trans-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD; 10-200 microM) had no effect on [Ca2+]i. Hill coefficients from curves fitted to concentration-response data suggested an amplification of the Ca2+ signal that was interpreted as calcium-induced calcium release (CICR) from intracellular Ca2+ stores. Caffeine (10 mM) produced a rapid transient rise in [Ca2+]i, confirming the existence of a Ca(2+)-sensitive store. Following caffeine-induced depletion of Ca2+ from intracellular stores, agonists were still able to produce increases in [Ca2+]i, confirming Ca2+ influx through the agonist-gated channel. The agonist-induced increase in [Ca2+]i was decreased following caffeine-induced depletion, confirming a process of CICR. These results are consistent with the hypothesis that excitatory amino acids can produce direct modulation of [Ca2+]i by influx through the agonist-gated channel and by CICR from intracellular stores.     

Noradrenergic modulation of the masseteric reflex in behaving cats. I. Pharmacological studies

The masseteric (jaw closure) reflex was utilized as a model system for assessing functional changes in central norepinephrine (NE) neurotransmission. This monosynaptic reflex was chosen because of its simple and well-defined circuitry, and because its motor component receives a dense NE innervation. Previous experiments in our laboratory described NE modulation of this reflex in the anesthetized rat. The present experiments examine the effects of NE on this response in the unanesthetized, behaving cat. The masseteric reflex was elicited by electrical stimulation of the mesencephalic trigeminal nucleus, and the response was recorded via electrodes permanently implanted in the masseter muscle. The amplitude of the reflex response was measured before and at various intervals following microinfusion (0.5 microliters) of NE or of various NE agonists directly into the motor trigeminal nucleus (MoV). Microinfusions of NE (0.125-5.0 micrograms) produced dose-dependent increases in the amplitude of the elicited reflex response. These effects were evident within 1 min postinfusion and lasted up to 30 min; in all cases, the response amplitude returned to baseline levels. The increase seen in response to 0.5 micrograms NE was blocked by pretreatment with the alpha-1-adrenergic antagonist prazosin, but not by pretreatment with the serotonin (5-HT) antagonist methysergide. Methysergide did, however, completely block the increase in the amplitude seen in response to microinfusion of 5-HT. Infusion of the alpha-1-adrenergic agonist phenylephrine also increased the amplitude of the reflex response. By contrast, infusion of the beta-adrenergic agonist isoproterenol had no effect, whereas clonidine, a presynaptic alpha-2-adrenergic agonist, decreased its amplitude.(ABSTRACT TRUNCATED AT 250 WORDS)