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Biomedical subjects

E Heilbronn

Publications and source records attributed to E Heilbronn.

At least 19 recordsLinked to original sources
PubMed1997

Uridine nucleotide receptors and their ligands: structural, physiological, and pathophysiological aspects, with special emphasis on the nervous system.

This review presents data on metabotropic uridine nucleotide receptor subtypes (P2UR) activated by UTP, sometimes also by UDP and/or ATP. Some chemical details of receptor subtypes and ligand interactions are described. Ligand-activated P2UR subtypes may couple to different second messengers, yet little is known about the nature of the coupling G-proteins. Data evaluating UTP as a physiological ligand include UTP origin, release and metabolism and illuminate especially roles for P2UR in the nervous system. No evidence shows UTP as a synaptic transmitter; sympathetic neurons may, however, carry P2UR allowing UTP-stimulation of norepinephrine release. UTP and derivatives act as therapeutic agents in several diseases involving mutated genes of transepithelial conductance regulators, including cystic fibrosis. This focuses interest to the synthesis of new compounds. Further, therapeutically used pyrimidine and pyrimidine analogues are suspected to have CNS-pathological effects. The presently scarce information in these areas strongly underlines the need for and importance of intense research on the suspected pyrimidine derivative triggered pathology as well as on the role of P2UR receptors in physiology and pathophysiology.

Animals↗
PubMed1996

Peptidyl conjugates of adenosine 5'-carboxylic acid synthesized and evaluated as ligands for P2 purinoceptors.

The role of extracellular ATP in vivo and the various cellular responses mediated by P2 purinoceptors have not yet been fully elucidated, in part depending on the lack of subtype-specific high affinity antagonists. Here we describe the synthesis of a new class of compounds, peptidyl derivatives of adenosine 5'-carboxylic acid, among which some have inhibitory effects in certain P2 purinoceptor-carrying biological systems, e.g., glioma and smooth muscle cell lines and isolated smooth muscle tissue preparations from guinea pig vas deferens and urinary bladder.

Adenosine↗
PubMed1995

Ligand-binding properties of an unusual nicotinic acetylcholine receptor subtype on isolated outer hair cells from guinea pig cochlea.

Acetylcholine receptors on isolated guinea pig cochlear outer hair cells (OHC) were characterized by radioligand binding. Equilibrium binding of [125I]alpha-bungarotoxin revealed a KD of 62 +/- 2 nM, Bmax = 7.2 +/- 1.8 x 10(7) binding sites/OHC, and a slowly reversible dissociation rate constant, kappa-1 = 2.2 +/- 0.01 x 10(-4) min-1. L-[3H]Nicotine bound reversibly (estimated KD approximately 230 nM and Bmax approximately 5 x 10(7)) with kinetic rate constants of association kappa-1 = 6.2 +/- 0.06 x 10(4) min-1 nM-1 and dissociation kappa-1 = 0.23 +/- 0.003 min-1. [3H]Strychnine bound to OHC with a KD of 35 +/- 6 nM and Bmax = 2.6 +/- 0.5 x 10(7), and binding increased 3-4 fold after membrane depolarization with 56.2 mM [K+], suggesting additional binding sites. Binding, seen only at > nM concentrations, of [3H]3-quinuclidinyl benzilate (KD = 11.5 +/- 5 nM; Bmax = 2.5 +/- 0.6 x 10(6)) was competitively inhibited by the muscarinic antagonists atropine and 4-DAMP (IC50 of 6.1 +/- 0.5 and 6.5 +/- 0.4 nM). The OHC receptor is thus an atypical nicotinic acetylcholine receptor subtype with unusual pharmacological properties.

Animals↗
PubMed1994

ATP-induced cytoplasmic [Ca2+] increases in isolated cochlear outer hair cells. Involved receptor and channel mechanisms.

Outer hair cells (OHC) of the mammalian cochlea are thought to preprocess the sound signal by active movements, which can be induced by electrical or chemical stimulation, e.g. depolarization evoked by high [K+] or increased cytoplasmic [Ca2+]. Extracellular ATP has been found to induce cytoplasmic [Ca2+] increases in OHC but involved mechanisms have not been elucidated. Cytoplasmic [Ca2+] was measured in non-enzymatically isolated single OHC using Fura-2 microspectrometry. Results, using ATP/derivatives and other P2-purinergic receptor (P2R) ligands, as well as Ca(2+)-channel blockers and pertussis toxin, revealed several signal transduction pathways that increase cytoplasmic [Ca2+] in OHC: a P2-purinergic receptor (P2R)--G-protein--effector (phospholipase C or an ion channel) system and a voltage-dependent Ca2+ channel. Agonist potency studies denote a pattern analogous to that found in skeletal muscle, i.e. ATP-alpha-S > ATP = 2-methyl-S-ATP >> ADP > alpha,beta-methylene-ATP, but no activation by ADP beta F or UTP, leaving a choice of P2y or P2zR subtypes. The latter possibility gained strength from calculations showing that up to 8% of ATP may have formed the P2zR agonist ATP4- in the experimental medium. Experiments in Ca(2+)-free medium and with pertussis toxin revealed that the main Ca2+ source was intracellular. Pertussis toxin did not affect [Ca2+] increase induced by carbachol. Acetylcholine, administered a few seconds before ATP, did not affect total cytoplasmic [Ca2+] increases. Induced cytoplasmic [Ca2+] increases were high enough (> 500 nM at 50 microM ATP/derivatives) to hyperpolarize the OHC membrane by opening K(+)-channels and decreased little with time.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1994

A new class of compounds, peptide derivatives of adenosine 5'-carboxylic acid, includes inhibitors of ATP receptor-mediated responses.

A new type of ligand for the study of P2-purinergic receptor subtypes was synthesized by combining and modifying conventional nucleoside chemistry with Fmoc solid phase peptide synthesis techniques. The tri- and tetra-aspartic acid derivatives of adenosine-5'-carboxylic acid (AdoCAsp3 and AdoCAsp4) were found to act as weak agonists at P2-purinergic receptors, (activated by ATP and UTP respectively) present on C6 glioma cells. AdoCAsp4 induced inositol 1,4,5-trisphosphate formation in the C6 cells with an EC50 of 73 microM. In addition, AdoCAsp4 was found to inhibit (IC50 approximately 80 microM) ATP-induced cytosolic [Ca2+] transients in these glioma cells. The glycine derivative, AdoCGly, increased evoked release of noradrenaline from mouse vas deferens slices, probably due to the blockade of presynaptic P2-autoreceptors. The possibility that aspartic, glutamic or gamma-carboxyglutamic residues may be used to replace phosphate groups on an ATP receptor ligand, opens up new ways in ligand design.

Adenosine↗
PubMed1992

Tetrahydroaminoacridine and related compounds interfere with fura-2 and indo-1.

In earlier experiments, using a fluorimetric method (fura-2), we found what seemed to be a decreased cytoplasmic [Ca2+] in neuroblastoma cells when 1,2,3,4-tetrahydro-9-aminoacridine (THA) was added, but discovered by repeating our work under cell-free conditions, that THA affected the Ca(2+)-fura-2 signal. The present study aimed at investigating the previously observed interference of THA with fura-2. 1 mM THA completely inhibits fluorescence of fura-2 and another Ca(2+)-indicator, indo-1, 1 microM each (apparent IC50 = 40 microM), presumably by absorbing excitation light.

4-Aminopyridine↗
PubMed1992

Receptor alterations in manganese intoxicated monkeys.

The density of four different receptors and one marker of dopamine uptake sites were analyzed in monkey brains after manganese exposure (0.1 g manganese per month during 26 months, a dose comparable to that workers might inhale in dusty environments) by means of quantitative receptor autoradiography. The binding of 3H-mazindol to the dopamine uptake sites was reduced by 75% in both the head of the caudate nucleus and putamen, while it remained unchanged in the other regions analyzed. The binding of the D1 receptor ligand 3H-SCH 23,390 was reduced about 45% in the same areas as mazindol binding, while the density of D2 receptors was unaffected. The muscarinic acetylcholine receptors as well as GABAA receptors remained also unchanged in all brain areas analyzed after manganese exposure. Thus the dopaminergic neurons must be considered to be vulnerable to manganese concentrations attainable in the work environment. Our results also indicate that postsynaptic structures containing D1 receptors are sensitive while cells containing D2 receptors are either spared or compensated for by up-regulation of the number of receptors on remaining sites.

Animals↗
PubMed1992

Manganese induced brain lesions in Macaca fascicularis as revealed by positron emission tomography and magnetic resonance imaging.

A series of positron emission tomography scans was made on two monkeys during a 16-month period when they received manganese(IV)oxide by subcutaneous injection. The distribution of [11C]-nomifensine uptake, indicating dopamine terminals, was followed in both monkey brains. The brain distributions of [11C]-raclopride, demonstrating D2 dopamine receptors, and [11C]-L-dopa, as a marker of dopamine turnover, were followed in one monkey each. The monkeys developed signs of poisoning namely unsteady gait and hypoactivity. The [11C]-nomifensine uptake in the striatum was reduced with time and reached a 60% reduction after 16 months exposure. This supports the suggestion that dopaminergic nerve endings degenerate during manganese intoxication. The [11C]-L-dopa decarboxylation was not significantly altered indicating a sparing of [11C]-L-dopa decarboxylation during manganese poisoning. A transient decrease of [11C]-raclopride binding occurred but at the end of the study D2-receptor binding had returned to starting values. The magnetic resonance imaging (MRI) revealed that the manganese accumulated in the globus pallidus, putamen and caudate nucleus. There were also suggestions of gliosis/edema in the posterior limb of the internal capsule. MRI might be useful to follow manganese intoxication in humans as long as the scan is made within a few months of exposure to manganese, i.e. before a reversal of the manganese accumulation.

Animals↗
PubMed1991

A nicotinic acetylcholine receptor-like alpha-bungarotoxin-binding site on outer hair cells.

Acetylcholine (ACh) appears to be the major neurotransmitter liberated from olivocochlear efferents terminating on outer hair cells (OHC). Recently, cholinergic receptor epitopes were visualized at the basal pole of the OHCs. To evaluate the ACh receptor type at OHC we performed binding studies with [125I]-labelled alpha-bungarotoxin (alpha-bgtx), a close to irreversibly acting blocker of the nicotinic acetylcholine receptor (nAChR) of skeletal muscle and of electrocytes of Torpedo and Electrophorus. An irreversible and saturable binding (80 nM) of the radiolabelled compound to OHCs was observed. The number of alpha-bgtx sensitive binding sites present on each OHC was calculated to be about 2 X 10(-17) mol/OHC, which would amount to about 10(7) binding sites/cell. Preincubation with the reversibly acting cholinergic ligands, carbamylcholine (1 mM), nicotine (0.1 mM) and d-tubocurarine (1-100 microM) was found to inhibit alpha-bgtx binding to a varying degree. Atropine (0.05 mM), a muscarinic antagonist, had no influence on the binding of alpha-bgtx to OHCs. [3H]-QNB, a specific marker and antagonist for muscarinic AChR, and [125I]-kappa-toxin, known to react with neuronal and ganglionic nAChR, showed no specific binding to OHCs. The data indicate that a peripheral type nAChR is present on OHCs mediating ACh-induced modulation of the biomechanics of the cochlea by influencing OHC motility.

Animals↗
PubMed1990

Receptor-triggered polyphosphoinositide turnover produces less cytosolic free calcium in cultured dysgenic myotubes than in normal myotubes.

Myotubes prepared from mice with muscular dysgenesis (mdg) were used to further elucidate the putative role of inositol triphosphate (InsP3) in excitation-contraction (E-C) coupling of skeletal muscle. The mdg mutation is characterized by an uncoupling of the E-C coupling. InsP3 production in normal and mdg/mdg myotube cultures and its relation to the levels of cytosolic free calcium were analyzed. Basal and ATP-stimulated levels of InsP3 were equal in normal and mdg/mdg myotube cultures. In contrast, the transient increases of cytosolic free calcium in mdg/mdg myotubes in culture were generally much lower than those in normal ones. This suggests that the defect in dysgenic myotubes does not rest on the InsP3 formation but on the InsP3-triggered transduction of excitation and/or the induction of calcium release from internal stores.

Adenosine Triphosphate↗
PubMed1989

Extracellularly applied ATP alters the calcium flux through dihydropyridine-sensitive channels in cultured chick myotubes.

Extracellularly applied ATP mediates a biphasic calcium signal in cultured chick myotubes. A rapid and transient increase in cytosolic calcium was independent of extracellular calcium while a second signal, slower in onset and decay, was absent without extracellular calcium. In depolarized myotubes, the cytosolic [Ca2+] was increased more than ten times above baseline level. Addition of ATP to the incubation medium immediately increased the rate of return of cytosolic Ca2+ levels to baseline. The ATP effect was half-maximal at about 10 microM ATP and was mimicked by ATP S. This ATP-sensitive calcium influx was also rapidly stopped by addition of dihydropyridines such as PN 200-110, suggesting that it is the voltage operated Ca2+-channel that was inactivated by ATP.

Adenosine Triphosphate↗
PubMed1989

Action potentials of cultured human oat cells: whole-cell measurements with the patch-clamp technique.

Oat cells (of the small cell carcinoma of the lung) have been reported to generate calcium action potentials. The calcium channels have further been suggested to play a crucial role in the relation between oat cell carcinoma and the often associated myasthenic syndrome. We have examined cultured human oat cells (U-1690) under voltage-clamp conditions, using the patch-clamp technique. We found, contrary to previous reports, that the action potential was caused by sodium and potassium currents. No calcium current was detected under these conditions, which indicates that calcium channels, if present, are very rare. The findings restrict, but do not rule out, the hypothesis that calcium plays a key role in the carcinoma/myasthenic syndrome relation.

Action Potentials↗
PubMed1988

P2-purinoceptor-stimulated phosphoinositide turnover in chick myotubes. Calcium mobilization and the role of guanyl nucleotide-binding proteins.

ATP, a trigger of P2-purinoceptor-mediated polyphosphoinositide (PI) turnover in cultured myotubes, increased cytosolic calcium levels in a time- and dose-dependent manner (quin2 fluorescence). The calcium was released from intracellular stores, as acute addition of 5 mM EGTA was without significant effect. Adenosine 5'-(3-thiotriphosphate) and 5'-adenylyl imidodiphosphate also increased intracellular levels of inositol phosphates (InsP) and cytosolic calcium levels. Treatment with cholera or pertussis toxin of myotube cultures did not affect the P2-purinoceptor-mediated InsP increase although PI turnover in permeabilized myotubes was stimulated by guanosine 5'-(3-thiotriphosphate). The results suggest that myotube P2-purinoceptors trigger PI turnover and increase intracellular free calcium levels, via a mechanism insensitive to ADP-ribosylation, by cholera or pertussis toxin of guanyl nucleotide-binding (G) proteins. However, the presence of a phospholipase C-coupled G-protein was otherwise demonstrated.

Adenosine Triphosphate↗
PubMed1987

Externally applied adenosine-5'-triphosphate causes inositol triphosphate accumulation in cultured chick myotubes.

In striated muscle, adenosine-5'-triphosphate (ATP) potentiates the responses to acetylcholine. The underlying biochemical events are unknown. Here we report that ATP, externally applied to chick myotubes, induces a rapid, dose-dependent accumulation of intracellular inositol triphosphate which is correlated with a decrease in phosphatidyl 4,5-bisphosphate. Adenosine-5'-diphosphate, adenosine-5'-monophosphate and adenosine are less potent while beta, gamma-imido ATP is equipotent motoneurons and/or skeletal muscle controls the activation of a polyphosphoinositide phosphodiesterase via a cell membrane P2-purinoceptor, thus modulating skeletal muscle responses to transmitter release.

Acetylcholine↗