Development of beta-adrenergic receptors and the in vitro accumulation of cyclic AMP in the chick spinal cord

To clarify the functional development of the descending monoaminergic input to the chick spinal cord we have studied the ontogeny of beta-adrenergic receptors by measuring the specific binding the tritiated dihydroalprenolol (DHA). In addition, we examined the ability of isoproterenol to stimulate the accumulation of cyclic AMP in slices of developing chick spinal cord. Results show that the chick spinal cord contains a high density of beta-adrenergic receptors that are apparently linked to adenylate cyclase. During development, both the density of beta-receptors, as determined by the specific binding of DHA, and the response of tissue slices to isoproterenol underwent marked changes. beta-Adrenergic receptors (approximately 4 fmol/mg tissue) were first detected on the fourteenth day in ovo. Receptor density increased to approximately 20 fmol/mg by day 20. Between day 20 and the time of hatching, a sharp increase in receptor density, to approximately 50 fmol/mg, was seen. The density of receptors remained high until the second day after hatching, fell off to approximately 30 fmol/mg by the fourth day, and remained relatively unchanged through day 30. The response of spinal cord slices to isoproterenol showed a similar pattern of development with the peak response (7-fold increase in levels of cyclic AMP) occurring at or near the time of hatching. During the period between day 18 in ovo and the time of hatching, when both the response of tissue slices to isoproterenol and the density of beta-receptors increased markedly, the activity of phosphodiesterase did not change. Therefore, the pronounced changes in adrenergic responsiveness that occurred near the time of hatching appear to be related primarily to changes in the density of beta-adrenergic receptors coupled to adenylate cyclase. Such developmental changes in the density of beta-adrenergic receptors and adrenergic responsiveness may play an important role in determining the functional state of the descending monoaminergic systems in the chick spinal cord.     

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)     

Characterization and measurement of [125I]iodopindolol binding in individual rat pineal glands: existence of a 24-h rhythm in beta-adrenergic receptor density

A simple procedure has been developed that permits measurement of beta-receptors in membrane preparations from individual rat pineal glands using [125I]iodopindolol ([125I]PIN). [125I]PIN binding to pineal membranes was stereospecific and saturable. Scatchard analysis of saturation isotherms yielded a Kd of 147.3 +/- 54 pM and a Bmax of 11.1 +/- 1.5 fmol/pineal gland. Binding was linear suggesting that [125I]PIN binds to a single population of pineal beta-adrenergic receptors. This procedure was used to evaluate 24-h variations in density of pineal [125I]PIN binding sites in male rats maintained in a 14:10 h light:dark cycle. Binding remained uniformly low during the daytime, increased slightly prior to lights off and peaked after 6 h of darkness decreasing abruptly 2 h later, before lights on. In animals maintained in light at night, the number of binding sites also increased, but did not exhibit the darkness-related decrease. The results demonstrate that beta-adrenergic receptors defined via [125I]PIN binding can be measured in tissue samples equivalent to less than one pineal gland. Moreover, the technique can be used in studies concerning the noradrenergic regulation of pineal function.     

Calcium and norepinephrine levels of rat salivary glands after terbutaline, dobutamine or isoproterenol

Present data show that a dose-related increase in calcium concentration [Ca] of submandibular glands of rat occurred after 6 days of twice daily administration of 25, 50 or 75 mg/kg b. wt. doses of the beta 1-adrenergic agonist, dobutamine, or the beta 2-adrenergic agonist, terbutaline. The beta 1-adrenergic receptor was responsible for mediation of these changes with both agonists since the effects of either agonist were prevented when a 10 mg/kg dose of the beta 1-antagonist, atenolol, was injected 20 min prior to the agonist, and the increase induced by either agonist was not prevented when the beta 1-adrenergic antagonist, butoxamine, was given prior to each agonist. Dobutamine caused more marked increases in [Ca] than did terbutaline. The parotid gland, however, showed a decrease with both agonists, and that caused by dobutamine was greater than that caused by terbutaline. Neither antagonist had any effect on the agonist-induced changes in the parotid gland. Reasons for the differences in response of the two glands are suggested. It does not appear, however, that, as suggested previously, the increase in the [Ca] of the submandibular gland is dependent on depletion of glandular levels of norepinephrine (NE). Of the agonists, only dobutamine caused a decrease in glandular concentration of NE, as well as total NE of the gland. With isoproterenol (ISO), NE concentration of parotid and submandibular was reduced, but with terbutaline only that of parotid was reduced. changes in total glandular NE were not found in either gland with either ISO or terbutaline. Thus, the decrease in concentration was a consequence of the increased mass of gland. With cyclocytidine (CC), both NE concentration and total NE were reduced, even though gland size increased. With reserpine (RES), NE concentration as well as total NE were markedly reduced (87-90%), and no change in gland size occurred. It is suggested, on the basis of present data, that prolonged activation of beta 1-adrenoceptors is the cause of the calcium accumulation, and that reduction in NE is not the cause of the calcium increase, but may only be a coincident event. Even with sympathectomy (here induced by reserpine), activation of beta-receptors over a long period of time is suggested as the cause of the calcium change, not the depletion of NE. The present data also provide the first evidence that CC causes NE depletion.     

A critical period of the development of beta-adrenergic receptor binding in the visual system of rat during visual deprivation

The density of [3H]dihydroalprenolol binding to beta-adrenergic receptors in the visual structures (visual cortex, superior colliculus and lateral geniculate nucleus) of rats raised under a normal 12 hr light-dark cycle was compared to those of rats visually deprived at different postnatal ages. Unilateral eyelid suture from postnatal days 10 or 16 to 3 months resulted in an increased [3H]dihydroalprenolol bilateral binding in the lateral geniculate nucleus compared to control animals. Monocular deprivation from postnatal days 25, 40, 60 and 90 had no effect on the density of [3H]dihydroalprenolol binding. After re-opening of the eyelid, which was sutured on postnatal day 10, at postnatal day 25 no changes in beta-adrenergic receptor binding in the lateral geniculate nucleus of the adult animal could be detected. After re-opening of the sutured eyelid on day 90, followed by examination of the adrenoceptor density 4 weeks later, the [3H]dihydroalprenolol binding in both lateral geniculate nuclei remained elevated as was also found in corresponding regions of monocular deprived animals. Binocular visual deprivation from postnatal day 10 until the age of 3 months had no effect on [3H]dihydroalprenolol binding in the visual centres in comparison to corresponding control animals. The data suggest that there exists a critical period for the ontogenetic development of beta-adrenergic receptors binding in the visual system of rats during which permanent alterations of receptor binding can be induced by monocular but not binocular visual deprivation.     

Identification and characterization of muscarinic cholinergic receptors in the human urinary bladder and parotid gland

The binding characteristics of [3H]quinuclidinyl benzilate (QNB) to muscarinic sites in isolated plasma membrane fractions of the human urinary bladder and parotid gland were studied. QNB binding to both preparations was of high affinity and low capacity. Mean values for the apparent dissociation constants (Kd) for binding to membrane preparations from the urinary bladder and parotid glands were 22 and 34 pM and the Bmax values 234 and 456 fmol/mg protein, respectively. Significance of difference between Kd and Bmax values from the two tissues was at the level of P less than 0.005 and P less than 0.05, respectively. QNB binding was inhibited by muscarinic receptor antagonists with varying degree of effectiveness. The mean values for the inhibition constant (Ki) were significantly lower for oxybutynin, amitriptyline, and pirenzepine but higher for secoverine in preparations of the urinary bladder than of the parotid gland. The mean Ki values for quinidine and verapamil were lower in the urinary bladder than that in the parotid gland. Carbachol exhibited a marked selectivity for the urinary bladder (about 30-fold) compared with the parotid gland. The present data obtained in two human tissues that are highly cholinergic in their innervation give support to the argument for heterogeneity of the muscarinic cholinergic receptors.     

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.     

Beta-receptor-stimulated and cyclic adenosine 3',5'-monophosphate-mediated taurine release from LRM55 glial cells

Adrenergic stimulation of LRM55 glial cells results in the release of the neuroactive amino acid taurine. The present study characterizes the receptors involved in taurine release and shows that taurine release is mediated by cyclic adenosine 3',5'-monophosphate (cAMP). beta-Receptors in LRM55 cells were first characterized by [125I]iodohydroxybenzylpindolol binding. Binding was stereospecific and saturable with time and ligand concentration. Kinetic analysis of equilibrium binding at 37 degrees C revealed a single component of high affinity (Km = 113 pm; Bmax = 52.1 +/- 5.0 fmol/mg of protein). The pharmacologies of the stimulation of cAMP accumulation and taurine release were similar. The agonists isoproterenol (IPR), epinephrine (E) and norepinephrine (NE) showed a rank order of potency characteristic of a beta-adrenergic system (IPR greater than E greater than or equal to NE). The beta-antagonists alprenolol and propranolol inhibited the IPR stimulation of both processes; the alpha-antagonist phentolamine did not. The dependence of taurine release on cAMP was further suggested by the similarity of the two time courses and was demonstrated by the stimulation of taurine release by the cAMP analogue dibutyryl cAMP. Thus, one physiological response of glial cells to beta-adrenergic stimulation is the release of taurine. Receptor-activated release of taurine from glia represents a previously undescribed neuronal-glial interaction by which glia may actively regulate neuronal excitability.     

Adrenergic and VIP stimulation of cyclic AMP accumulation in ovine pineals

In vitro autoradiography of [125I]cyanopindolol ([125I]cyp) binding to sections of ovine pineal reveals a uniform distribution of beta-adrenergic receptors throughout the gland. Norepinephrine (NE) stimulated cyclic AMP production in pineal slices in both a time- and dose-dependent manner, producing a maximal two-fold increase. NE, isoproterenol (ISO) and epinephrine (E) stimulate cyclic AMP (cAMP) production with equal potency. NE stimulation of cAMP was totally blocked by propranolol (beta antagonist) but only partially blocked by practolol (a beta 1 preferential antagonist) indicating a mixed population of beta 1 and beta 2 receptor subtypes. Displacement of [125I]cyp binding by either practolol or zinterol (preferential beta 2 agonist) revealed IC50s of 1.3 x 10(-5) M and 9.95 x 10(-8) M respectively, confirming a mixed population of beta 1 and beta 2 receptors. A range of peptides previously localised within the pineal by immunocytochemistry were tested at a concentration of 10(-5) M for their effect on cyclic AMP production in pineal homogenates. Only vasoactive intestinal peptide (VIP) was effective showing a dose-dependent stimulation. ISO and VIP stimulation of cAMP were additive indicating action via independent receptors.     

Autonomic regulation of cystatin S gene expression in rat submandibular glands

Innervation of rat submandibular and parotid glands by the autonomic nervous system regulates saliva volume, its rate of secretion and its composition. The autonomic nervous system also plays a regulatory role in the differentiation and growth of salivary glands, and in the expression of specific sets of genes. Rat cystatin S, a member of family 2 of the cysteine proteinase inhibitor superfamily, is expressed in submandibular and parotid glands of human and rat. In the rat, cystatin S gene expression is tissue- and cell type-specific, is temporally regulated during postnatal development, and not observed in adult animals. The beta-adrenergic agonist isoproterenol (IPR) induces hypertrophic and hyperplastic enlargements of rat salivary glands and the expression of a number of genes including cystatin S. Sympathectomy reduces, but does not completely block, IPR-induced expression of the cystatin S gene in submandibular glands of adult female rats, indicating the participation of sympathetic factor(s) in its regulation. Bilateral parasympathectomy also reduces IPR-induced cystatin S gene expression, suggesting a role of the parasympathetic nervous system in its regulation. Experiments described in this paper suggest that similar factor(s) arising from both the sympathetic and parasympathetic branches of the autonomic nervous system simultaneously participate in IPR-induced cystatin S gene expression in submandibular glands.     

Increased serotonin2 and beta-adrenergic receptor binding in the frontal cortices of suicide victims

A statistically significant 28% increase in the mean (+/- SD) number of serotonin2 receptors (127.8 +/- 13.4 vs 99.6 +/- 11.1 fmol/mg of protein) and a 73% increase in beta-adrenergic receptor binding (14.5 +/- 1.5 vs 8.4 +/- 1.5 fmol/mg) was found in the frontal cortices of violent suicide victims compared with matched controls. No significant differences were found in the number of serotonin1 binding sites (109.5 +/- 13.4 vs 99.9 +/- 8.8 fmol/mg). We have previously reported a reduced density of presynaptic tritiated imipramine binding sites on serotonergic nerve terminals in the frontal cortices of suicide victims. These data support the hypothesis that suicide completed by violent methods is associated with reduced presynaptic serotonergic activity that has generated compensatory upregulation of the postsynaptic serotonin2 receptor sites. The increase observed in beta-adrenergic binding suggests that there may also be a concomitant reduction in presynaptic noradrenergic activity associated with suicide. If antidepressant pharmacotherapies specifically downregulate cortical beta-adrenergic and/or serotonin2 receptors in depressed subjects, as has been demonstrated in animal studies, and since these effects would be in the opposite direction of the receptor changes found in suicide victims, they may account for the therapeutic action of antidepressants on suicidal behavior and depressive disorders.     

Interaction between alpha 2- and beta-adrenergic receptors in rat cerebral cortical membranes: clonidine-induced reduction in agonist and antagonist affinity for beta-adrenergic receptors.

The interaction between alpha 2- and beta-adrenergic receptors was investigated in rat cerebral cortical membranes. Clonidine inhibition of [3H]dihydroalprenolol ([3H]DHA) binding resulted in biphasic competition curves with a mean Hill coefficient of 0.45. The addition of 1 microM yohimbine caused a rightward shift of the first portion of the clonidine inhibition curve. In the presence of 1 microM clonidine, the maximum concentration which did not inhibit [3H]DHA binding, inhibition curves of [3H]DHA binding by isoproterenol shifted to the right. A mean Hill coefficient increased from a control value of 0.63 to 0.76. Computer modeling analysis revealed that 1 microM clonidine decreased a beta-adrenergic high-affinity state from 28% to 13%. However, the addition of 1 microM yohimbine completely prevented the clonidine-induced reduction in the beta-adrenergic high-affinity state. In the presence of 200 microM GTP, the effect of clonidine was not observed. In addition, Kd and Bmax values for [3H]p-aminoclonidine ([3H]PAC) binding were not significantly changed by the addition of 100 nM isoproterenol, the maximum concentration which did not inhibit [3H]PAC binding. Moreover, isoproterenol inhibition of [3H]PAC binding resulted in steep competition curves with a mean Hill coefficient of 0.97. The addition of 1 microM alprenolol did not affect the isoproterenol inhibition curve. These data demonstrated that clonidine caused a decrease in agonist and antagonist affinity for beta-adrenergic receptors, while isoproterenol did not modulate the binding characteristics of alpha 2-adrenergic receptors. Furthermore, these results suggest that regulation between alpha 2- and beta-adrenergic receptors is not bidirectional, but is instead unidirectional from alpha 2-adrenergic receptors to beta-adrenergic receptors.     

Electrophysiological responsiveness to isoproterenol in rat hippocampal slices correlates with changes in beta-adrenergic receptor density induced by chronic morphine treatment

The effects of chronic morphine treatment and morphine withdrawal on beta-adrenergic receptor density and electrophysiological responsiveness in rat hippocampus were examined. Chronic treatment of rats with morphine for 14 days resulted in a 19% increase in the number of beta-adrenergic receptors in hippocampus, as measured by the binding of the specific antagonist [3H]dihydroalprenolol (DHA). In comparison, the number of specific binding sites for [3H]DHA was decreased 27% in hippocampus in morphine-withdrawn animals, compared to saline-treated controls. These alterations in beta-adrenergic receptor density were not accompanied by a significant change in the dissociation constant (Kd) for [3H]DHA or in the inhibitory constants (Ki) for the displacement of the [3H]-antagonist by either norepinephrine or isoproterenol. Electrophysiological experiments in the in vitro hippocampal slice preparation revealed that responses to threshold as well as maximal concentrations of isoproterenol were significantly enhanced in morphine-dependent animals, compared to controls, whereas electrophysiological responsiveness to maximal concentrations of isoproterenol was decreased in slices from morphine-withdrawn rats. The results of this study indicate that beta-adrenergic receptors in hippocampus are up-regulated during the development of morphine dependence and down-regulated during opiate withdrawal. These changes in hippocampal beta-adrenergic receptor density are likely to be of functional relevance since they are manifested in a corresponding increase and decrease, respectively, in electrophysiological responsiveness to an exogenously administered beta-adrenergic receptor agonist.     

Daily changes in beta-adrenergic sensitivity of rat submandibular gland. Correlation with beta-adrenoceptor rhythm

Neurons from the superior cervical ganglia (SCG) innervate the submandibular gland and release noradrenaline during the dark phase of the daily photoperiod. Since in the pineal, another structure innervated by sympathetic neurons, nocturnal activation of the SCG is associated with beta-adrenergic sub- and super-sensitivity rhythms, the possible existence of similar phenomena in the rat submandibular gland was assessed. Wistar female rats, kept on a 14:10 light/dark cycle (light from 06:00 to 20:00 h), were sacrificed at 09:00, 14:00, 20:00, 24:00 and 04:00 h. beta-Adrenoceptors were studied by 3H-dihydroalprenolol binding to membrane preparations. The equilibrium dissociation constant (Kd) did not change as a function of time while significant daily variations in maximal binding values (Bmax) were observed with a peak at 20:00 h. Changes in Bmax correlated with a high response of adenylate cyclase to isoproterenol. In addition, when the response in salivary flow to isoproterenol was measured. a shift to the left (about 1 logarithmic unit) in dose-response curves was observed at 19:00-20:00 has compared to 08:00-09:00 h. These daily variations in isoproterenol responsiveness seem not to depend on the pattern of eating since a 24-h starvation or a nocturnal starvation for 16-18 days did not abolish the morning-evening differences in the salivary flow response to isoproterenol. Rather, the results suggest that the daily variations in isoproterenol response correlate with beta-adrenergic super- and sub-sensitivity phenomena associated with the circadian release of noradrenaline from SCG neurons.     

Human M1-, M2- and M3-muscarinic cholinergic receptors: binding characteristics of agonists and antagonists

The muscarinic acetylcholine receptors were identified in membrane preparations from human tissues by the specific binding of 1-[benzilic-4,4'-3H] quinuclidinyl benzilate. Saturation binding isotherms of this radioligand yielded a total amount of receptors of 435 +/- 208, 159 +/- 65 and 913 +/- 89 fmol/mg protein, respectively, in the hippocampus, pons and submandibular gland. Non linear least squares analysis of competition binding studies with the antagonists pirenzepine and AF-DX 116 indicates that the majority of receptors are of the M1-type in the hippocampus (83%, high affinity for pirenzepine, intermediate affinity for AF-DX 116), the M2-type in the pons (low affinity for pirenzepine and high affinity for AF-DX 116), and the M3-type in the submandibular gland (low affinity for pirenzepine and AF-DX 116). Competition binding parameters of the agonists carbachol, arecoline, oxotremorine, pilocarpine and MCN-A-343 were compared for M1, M2 and M3 receptors in the human hippocampus, pons and submandibular gland. GTP caused a shift to the right and a steepening of the shallow agonist competition curves in the 3 tissues but did not affect the initially steep ones. This effect is explained by a GTP-mediated conversion of high- to low-agonist affinity sites. The extent of the nucleotide shift was much greater for M2 receptors as compared with M1 and M3 receptors. The GTP effect was impaired by the sulphydryl reagent N-ethylmaleimide, probably due to alkylation of GTP-binding proteins. Moreover, the reagent provoked also an increase of the agonist affinity for the uncoupled muscarinic receptors. For all agonists, this increase was more pronounced for the M2 receptors than for the M1 and M3 receptors. These findings suggest structural differences between the agonist binding sites of M1 and M3 receptors versus the M2 receptors.     

Identification and localization of adrenergic receptors in cat visual cortex

The concentration and location of adrenergic receptors in cat visual cortex have been determined by radioligand binding techniques using [3H]prazosin (alpha 1-adrenergic receptors), [3H]yohimbine (alpha 2-adrenergic receptors) and [3H]dihydroalprenolol (beta-adrenergic receptors). Saturable high affinity binding sites for all of these ligands were found. The beta-adrenergic receptor population was resolved into beta 1- and beta 2-sites that were present in the ratio 35:65. The laminar distributions of the alpha 1-, alpha 2- and beta-adrenergic receptors were different. The alpha 1- and beta-adrenergic receptors were very similarly localized, being seen in upper layers (I, II and III) and lower layers (layers V and VI). The labelling in upper layers was greater than that in lower layers, more so for alpha 1-adrenergic receptors than beta-adrenergic receptors. alpha 2-Adrenergic receptors were seen in a single band that occupied layer II and III but did extend to the pial surface. These results indicate that the effect of norepinephrine on neuronal activity in cat visual cortex will depend upon the layer in which it is released. Our results provide a basis for further physiological studies of the role of norepinephrine in the processing of visual information.     

Effects of alpha, beta 1, and beta 2 adrenergic antagonists on the Na and K concentrations of sympathetic-nerve stimulated rat saliva

Selective alpha and beta 1 and beta 2 adrenergic antagonists were used with electrical stimulation of the sympathetic innervation to parotid and submandibular glands of rats in order to delineate the role of the beta 1 and beta 2 adrenoceptors in regulation of salivary flow rate, Na reabsorption and K secretion from these glands. In parotid gland, [Na] of sympathetically evoked saliva in the presence of phentolamine (3 mg/kg, i.p.) was not different from that of nerve-evoked saliva in the presence of phentolamine and butoxamine (3 mg/kg, i.p.), except for the last 20 min of stimulation when [Na] of nerve-elicited saliva was higher. [K] of saliva with sympathetically evoked stimulation was the same in the presence of phentolamine alone as it was or in the presence of phentolamine and butoxamine. Again, there was no difference in salivary flow rate induced by either kind of stimulation, except for the first 10 min of stimulation, during which salivary flow rate of nerve-evoked saliva in the presence of phentolamine was lower than under the other conditions indicated. On the other hand, with submandibular gland, [Na] and [K] of nerve-elicited saliva in the presence of phentolamine were generally higher than levels of sympathetically evoked saliva in the presence of phentolamine and butoxamine. However, salivary flow rate of nerve-evoked saliva in the presence of phentolamine was generally lower than that of sympathetically evoked saliva in the presence of phentolamine and butoxamine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Muscarinic receptors of rat parotid gland enlarged by gland ablation and bulk diet. Effects of denervation

The density of muscarinic binding sites was increased 10% in the rat parotid gland enlarged (2 times control) as a result of ablation of the submandibular-sublingual glands and maintenance of rats on bulk diet (50% inert cellulose plus 50% solid chow) for 4 weeks. When either the parasympathetic or sympathetic innervation to the gland was unilaterally removed at the time of submandibular-sublingual ablation and introduction of the bulk diet, the density of muscarinic receptors showed an even greater increase from levels of innervated glands of chow-fed controls (29%); with removal of both nerves, the increase was 39%. A 36% increase in cyclic guanosine monophosphate levels accompanied the increase in receptors of the enlarged gland, but when the parotid was denervated, there was no change in cyclic GMP. Absence of either or both nerves led to a maximal decrease of 24-29% in density of muscarinic receptors of parotid gland of chow-fed controls, but to no change in cyclic GMP levels. While autonomic influences mediate the changes in density of muscarinic receptors of parotid gland of chow-fed rats, some additional factor is apparently involved in their increase in the enlarged gland.     

Serotonin regulation of nerve growth factor synthesis in neonatal and adult astrocytes: comparison to the beta-adrenergic agonist isoproterenol

Although serotonin regulates synthesis of the neurotrophic factor S-100 beta by astrocytes, its ability to affect nerve growth factor (NGF) synthesis has never been examined. We report here that there is a correlation between the effect of serotonin on cyclic adenosine monophosphate (cAMP) content and on NGF content in neonatal astrocytes but not in adult astrocytes. In neonatal striatal astrocytes, serotonin increases both cAMP and NGF, whereas, in neonatal cerebellar astrocytes, serotonin decreases both. The increase in neonatal cortical astrocyte cAMP appeared to be too small (45%) to increase NGF significantly. The beta-adrenergic agonist isoproterenol increased cAMP and NGF in both cortical and striatal astrocytes derived from neonatal rats. In contrast, there was a dissociation between cAMP changes and NGF changes in astrocytes derived from adult rats. Both serotonin and isoproterenol increased cAMP in adult cortical astrocytes, without any effect on NGF content. However, adult striatal astrocytes responded to serotonin with an elevation of both cAMP and NGF, whereas isoproterenol could only enhance cAMP, without affecting NGF. Thus, in neonatal astrocytes, a change of sufficient magnitude in cAMP was correlated with a comparable change in NGF, in response to activation of either serotonergic or beta-adrenergic receptors; in cerebellar astrocytes, the decrease in cAMP was accompanied by a decrease in NGF. In contrast, adult astrocytes were not responsive: Although cAMP changes were large, NGF synthesis was increased only in striatal astrocytes and only in response to serotonin. J. Neurosci. Res. 64:261-267, 2001. Published 2001 Wiley-Liss, Inc.     

Effect of age on the rate of recovery of beta-adrenergic receptors in rat brain following desmethylimipramine-induced subsensitivity

Brain tissues from aged rats have an impaired ability to increase beta-adrenergic receptors in response to reduced noradrenergic input, but can down-regulate these receptors in response to repeated administration of desmethylimipramine (DMI). In this study we compared the ability of brain tissues from young (3-month) and aged (20- to 26-month) rats to restore their density of beta-adrenergic receptors following desmethylimipramine (DMI)-induced receptor subsensitivity. Either DMI or saline was administered i.p. twice daily for 7 days to groups of young and aged rats. At various times after drug administration [3H]dihydroalprenolol (DHA) binding was determined in homogenates of pineal gland and cerebral cortex. Four hours after the last dose of DMI there was a decrease in DHA binding in both brain areas of young and aged rats. In young rats DHA binding in these tissues returned to control levels by 2 days after DMI administration. In contrast, in aged rats it took 8 and 16 days for DHA binding to recover in cerebrum and pineal, respectively. The concentration and half-life for the disappearance of DMI from serum and cerebrum were significantly greater in aged rats than in young rats, but the differences do not entirely explain the delayed recovery of beta-receptors in the aged rats. The results suggest that beta-adrenergic receptors of brain tissues from aged rats cannot recover from beta-receptor subsensitivity as readily as those from young rats. If this recovery process requires the synthesis of new receptors, then this synthetic mechanism may be impaired with age.(ABSTRACT TRUNCATED AT 250 WORDS)