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R C Deth

Publications and source records attributed to R C Deth.

48 records · Page 3Linked to original sources
PubMed1985

Phosphoinositide hydrolysis is correlated with agonist-induced calcium flux and contraction in the rabbit aorta.

In the present study changes in the extent of 32P labelling of membrane phospholipids were correlated with the alpha 1-adrenoceptor-induced events of increased 45Ca influx, 45Ca release and contraction in the rabbit aorta. Under basal conditions 32P incorporation into all phospholipids proceeded without saturation through 80 min of labelling. During a 5 min exposure to 10(-5) M norepinephrine (NE) after 25 min of prelabelling the incorporation of 32P into certain phospholipids was substantially increased. Phosphatidic acid (PA) labelling was increased above basal levels by 4.1 fold, phosphatidylinositol (PI) 2.5 fold and phosphatidylcholine (PC) 1.8 fold. Half maximal stimulation of 32P labelling of PA occurred at 2.0 microM, which was similar to the EC50 value for stimulation of 45Ca influx (2.5 microM) and 45Ca release (2.1 microM) but slightly higher than the value for contractile response (0.9 microM). Antagonist sensitivity studies reinforced the alpha 1 receptor subtype character of the rabbit aorta. Prazosin (10(-7) M) reduced agonist-induced events by 63-82% while yohimbine (10(-7) M) was without influence. Phenoxybenzamine (10(-8) M) reduced agonist-induced events by 56-76%. A temporal comparison showed that agonist stimulation of PA labelling was slower than 45Ca release, but similar to the time course of 45Ca influx. Hydrolysis of 32P-labelled phosphatidylinositol diphosphate (PIP2) was more rapid and paralleled 45Ca release. These findings suggest that PIP2 hydrolysis may account for the rapid phase of norepinephrine-induced contraction in rabbit aorta while PA or its immediate precursor diacylglycerol may account for receptor-induced Ca2+ influx.

Animals↗
PubMed1985

Evidence for a decrease in the efficiency of beta-receptor coupling to adenylate cyclase in liver membranes from sucrose-fed rats.

Sucrose feeding has been shown previously to alter the plasma concentration of several factors which may regulate beta-adrenergic receptors, including corticosteroids and insulin as well as altered sympathetic nervous system (SNS) tone. For this reason we initiated a study of the effects of sucrose feeding on the beta-adrenergic receptor-adenylate cyclase system in rat liver plasma membranes. Beta-Adrenergic responsiveness was monitored by measuring isoproterenol stimulation of adenylate cyclase activity, while beta-adrenergic receptor characteristics were evaluated by analyzing [125I]iodocyanopindolol [( 125I]CYP) binding. Rats fed rat chow ad lib. supplemented by drinking water containing 10% sucrose solution exhibited a 50-75% reduction in hepatic isoproterenol-sensitive adenylate cyclase activity. This effect of sucrose was also observed in adrenalectomized (ADX) and 6-hydroxydopamine-pretreated animals, ruling out a causal role for corticosteroids or the sympathetic nervous system respectively. No effect was observed on basal, glucagon-, fluoride- or GTP-stimulated adenylate cyclase. A small but significant decrease in [125I]CYP specific binding capacity was observed in liver membranes prepared from sucrose-fed ADX rats, whereas no change in [125I]CYP binding capacity was observed in in sucrose-fed normal rats. These observations suggest that beta-receptor to adenylate cyclase coupling efficiency is decreased by the sucrose diet. The activities of two membrane-associated phospholipid methyltransferases and the content of endogenous S-adenosylmethionine in liver were reduced by sucrose feeding, implying a defect in the methylation pathway for phosphatidylcholine synthesis. The possible relationship between this latter finding and the observed decrease in beta-adrenergic receptor to adenylate cyclase coupling efficiency is discussed.

Adenylyl Cyclases↗
PubMed1984

Phorbol ester-induced contraction of arterial smooth muscle and inhibition of alpha-adrenergic response.

The influence of the phorbol ester, 12-o-tetradecanoylphorbol-13 acetate (TPA) on the contractility of rat and rabbit aorta was assessed. At 10(-8) to 10(-5)M TPA produced a gradual contraction of rat aorta, while rabbit aorta gave only a small contraction at 10(-5)M. Contractions were irreversible and dependent on extracellular Ca2+. Associated with TPA-induced contraction, alpha-adrenergic receptor responses of the rat were inhibited in a noncompetitive manner, while rabbit responses were little affected. Since TPA activates phospholipid-dependent protein kinase C in a manner analogous to endogenous diacylglycerol, a critical role for the latter in alpha-adrenergic events is suggested.

Animals↗
PubMed1984

Lack of correlation between [3H]ouabain binding and Na-K ATPase inhibition in rat aorta.

The binding of [3H]ouabain to intact strips of rat aorta was compared with the ability of ouabain to inhibit the uptake of 86Rb by the same preparation. When a cold temperature wash was used to process tissues after binding of [3H]ouabain, a class of relatively high affinity binding sites was found (KD = 1.2 X 10(-7) M). Binding was saturable and sensitive to both ATP depletion and elevated potassium. Elevation of cytoplasmic Ca2+ levels by phenylephrine or c-AMP levels by theophylline and terbutaline had no influence on [3H]ouabain binding. Ouabain inhibition of 86Rb uptake progressed to 60% of the total 86Rb uptake at 2 X 10(-3) from a threshold of about 10(-5) M. Half-maximal inhibition by ouabain occurred at a concentration of 10(-4) M. The disparity between [3H]ouabain binding and inhibition of 86Rb uptake indicates that the high affinity binding site in the rat does not contribute to inhibition of Na-K ATPase function.

Animals↗
PubMed1984

Effect of sucrose feeding on alpha 1-adrenergic responses in rat liver.

A sustained increase in sympathetic nervous system (SNS) activity was induced by substituting a 10% sucrose solution for the drinking water of rats fed laboratory chow ad libitum. The effects of increased SNS activity on alpha 1-adrenergic processes in liver were examined by evaluating three alpha 1-responses, namely, phenylephrine-stimulated ouabain-sensitive 86Rb+ uptake, 45Ca2+ efflux, and glucose release. Sucrose feeding abolished phenylephrine stimulation of ouabain-sensitive 86Rb+ uptake and 45Ca2+ efflux and induced a three- to fourfold reduction in the ability of phenylephrine to stimulate glucose release from liver slices. Pretreatment with 6-hydroxydopamine markedly reduced liver norepinephrine content. When 6-hydroxydopamine was used to prevent the sucrose-induced increase in SNS activity, the changes in 86Rb+ uptake, 45Ca2+ efflux, and glucose release that otherwise followed sucrose feeding were not observed. Sucrose feeding did not alter binding of the alpha 1-antagonist [3H]prazosin to liver cell membrane alpha 1-receptors or displacement of [3H]prazosin by the alpha-agonist epinephrine. These observations suggest that sustained increases in SNS activity may have profound effects on liver alpha 1-adrenergic events that occur subsequent to hormone-receptor interaction.

Animals↗
PubMed1984

Release of a common source of intracellular Ca2+ by alpha-adrenergic agonists and dinitrophenol in rat liver slices.

The effects of alpha-adrenergic agonists on 45Ca efflux from slices of rat liver were studied and the results compared to earlier studies on the rabbit aorta. Norepinephrine and phenylephrine (PE) cause a stimulation of 45Ca efflux which was due to alpha 1-receptor activation. The mitochondrial uncoupler dinitrophenol (DNP) also stimulated 45Ca efflux while caffeine had little or no effect. PE and DNP effects were due to the release of intracellular 45Ca stores, and both agents released a similar quantity of Ca2+. 45Ca/40Ca exchange in both PE- and DNP-released sources was similar (t1/2 approximately or equal to 16 min). DNP and PE effects were not additive, and previous exposure to one agent reduces the response to the other. These results suggest that alpha 1-agonists and DNP release a common source of intracellular Ca2+ in rat liver. Since in rabbit aorta alpha-agonists primarily release Ca2+ from a nonmitochondrial source, our results suggest alpha 1-receptors act via the generation of a Ca2+-releasing substance which subsequently mobilizes Ca2+ from different organelles in different tissues.

2,4-Dinitrophenol↗
PubMed1983

Simultaneous loss and reappearance of alpha 1-adrenergic responses and [3H]prazosin binding sites in rat liver after irreversible blockade by phenoxybenzamine.

The relative influences of the in vivo administration of phenoxybenzamine on in vitro binding to alpha 1-adrenergic receptors and alpha 1-receptor-mediated responses were studied. Phenoxybenzamine treatment reduced maximal specific binding of the alpha 1-selective antagonist [3H]prazosin to liver cell membranes. This response was rapid (less than 90 min) and half-maximal following a phenoxybenzamine dose of approx. 10 mg/kg. A similar decrease in the ability of phenylephrine to stimulate glucose release and 45Ca2+ efflux from liver slices was also noted after phenoxybenzamine treatment. During the recovery period following administration of 30 mg/kg phenoxybenzamine, [3H]prazosin specific binding and phenylephrine-stimulated glucose release and 45Ca2+ efflux returned to their respective control levels with t 1/2 values of 42, 49 and 38 h, respectively. At all times studied during the recovery period, alpha 1-binding and both of the alpha 1-responses were similar fractions of their respective control values. These observations indicate that a close relationship exists between the density of [3H]prazosin binding sites and the ability of rat liver to respond to alpha 1-stimulation. We suggest that the binding sites identified in studies using the antagonist [3H]prazosin and those through which the agonist phenylephrine stimulates glucose release and 45Ca2+ efflux are either identical or in equilibrium with each other.

Animals↗
PubMed1983

Alpha adrenergic receptor subtype associated with receptor binding, Ca++ influx, Ca++ release and contractile events in the rabbit aorta.

The alpha-adrenergic receptor mechanism in rabbit aorta was examined for the involvement of alpha-1 or alpha-2 receptor subtypes. Agonists (phenylephrine, norepinephrine and clonidine) and antagonists (prazosin and yohimbine) with known receptor subtype selectivity were used to define the contribution of alpha-1 or alpha-2 receptors to receptor-initiated cellular Ca++ influx, intracellular release of Ca++ and overall contraction. The receptor content of isolated membranes was also measured in [3H]prazosin and [3H]yohimbine radioligand binding studies. Contraction-derived KB values for prazosin (3 nM) or yohimbine (1 microM) were similar for all three agonists, indicating that each acted on the same alpha-1 receptor. Prazosin (10(-7) M) was effective in causing inhibition of cellular Ca++ influx initiated by agonists whereas yohimbine (10(-6) M) had no effect. Prazosin but not yohimbine caused a partial reduction in phenylephrine or norepinephrine-induced stimulation of 45Ca efflux rate whereas the smaller clonidine-induced stimulation was totally inhibited by prazosin and partially inhibited by yohimbine. Alpha-1 specific binding of [3H]prazosin was observed with a KD of 3.5 nM and maximum binding site (Bmax) of 73 fmol/mg of protein. Although no alpha-2 specific binding of [3H]yohimbine was observed, binding to a low-affinity/high-capacity class of sites was found. The results indicate the sole presence and contribution of alpha-1 receptors to Ca++ flux and contractile events in the rabbit aorta.

Animals↗
PubMed1981

Mobilization of a common source of smooth muscle Ca2+ by norepinephrine and methylxanthines.

The ability of norepinephrine (NE), caffeine (CAF), theophylline (THEO), dinitrophenol (DNP), and potassium (high K) to mobilize cellular Ca2+ in rabbit aorta was examined using 45Ca-efflux techniques. After 10 min of Ca2+ deprivation using either Ca-free EGTA or Ca-free lanthanum (La3+) buffers, NE, CAF, and DNP still caused an increase in 45Ca-efflux rate, indicating a cellular source of 45Ca, while high K did not. Contractile behavior after Ca removal paralleled 45Ca-efflux events. CAF (10 mM) inhibited NE contractile responses, and this inhibition was associated with the depletion of the NE-releasable Ca2+ store. Previous exposure to CAF during 45Ca efflux reduced subsequent stimulation of 45Ca efflux was not additive. THEO caused a stimulation of 45Ca efflux similar to CAF. CAF, THEO, and 3-isobutyl-l-methylxanthine caused a two- to threefold increase in cAMP levels in association with their stimulation of 45Ca efflux. These results suggest that NE and methylxanthines mobilize a common cellular Ca2+ source that is associated with contraction in the case of NE but relaxation in the case of methylxanthines.

Animals↗
PubMed1978

Effect of lanthanum and reduced temperature on 45Ca efflux from rabbit aorta.

The ability of lanthanum (La3+) to block calcium efflux from smooth muscle cells of the rabbit aorta has been examined. La3+ promotes the very early phase of 45Ca efflux, which is extracellular in origin, and partially inhibits the latter, cellular portion. Stimulation of 45Ca efflux caused by the release of intracellular 45Ca with either 10(-4) M dinitrophenol or 10 mM caffeine was not reduced by pretreatment with 10 mM La for 40 min, whereas stimulation due to norepinephrine was abolished. It was concluded that during the use of the "La method" for measuring cellular 45Ca there is an underestimation due to unblocked 45Ca loss. This loss can be reduced by processing tissue at 2 degrees C, which inhibits transport processes. The time course of 45Ca uptake and the stimulation of uptake by high K+ are qualitatively but not quantitatively similar if tissues are washed at 37 and 2 degrees C. Tissues washed in La3+ at 2 degrees C for 60 min retain approximately double the cellular 45Ca of those washed at 37 degrees C. This methodology provides an improved correlation between estimates of cellular calcium derived from 45Ca uptake and 45Ca efflux experiments.

Animals↗