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M Wüster

Publications and source records attributed to M Wüster.

At least 19 recordsLinked to original sources
PubMed1986

Target size analysis of opioid receptors. No difference between receptor types, but discrimination between two receptor states.

Target size analysis of opioid receptor is complicated by the presence of multi-exponential inactivation curves. Irradiation of intact frozen tissue proved essential to eliminate such artifacts, due to indirect irradiation effects. Upon irradiation condition, opioid binding activity was inactivated in a single mono-exponential manner. Identical inactivation curves were obtained for mu, delta and kappa binding activities in brain membranes from rat, guinea-pig and frog and in NG 108-15 cells: the molecular mass obtained was 98 +/- 2 kDa. However, when opioid binding was assayed in the presence of Na+, Mg2+ and GTP, the molecular mass was found to be only 56 +/- 4.4 kDa. We suggest that the opioid recognition site comprises a unit of 56 kDa and that in the absence of Na+, Mg2+ and GTP an additional membrane component of 40-44 kDa is necessary for high-affinity opioid binding.

Animals↗
PubMed1985

Specific changes in local cerebral glucose utilization in the rat brain induced by acute and chronic diazepam.

The effect of acute and long term administration of diazepam upon local cerebral glucose utilization (LCGU) in discrete regions of the central nervous system of the rat was studied by means of a quantitative autoradiographic [14C]2-deoxy-D-glucose technique. A single injection of diazepam (1.0 mg/kg and 2.5 mg/kg i.v.) reduced LCGU up to 30% in particular brain areas, such as the lateral and ventral thalamic nuclei, the medial geniculate body and the mamillary body, whereas the activity in many other structures was not significantly altered. That the effects may be mediated via specific benzodiazepine receptors was indicated by the ability of the selective benzodiazepine antagonist Ro 15-1788 (imidazobenzodiazepine) to attenuate these actions. Rats treated chronically with diazepam (20 mg/kg day for a period of 14 days) still displayed slight reductions of LCGU in certain areas affected acutely such as the lateral thalamic nucleus. In contrast to its effects in rats chronically treated with vehicle, an acute diazepam injection failed to significantly modify LCGU in rats chronically treated with diazepam. Furthermore, in chronic diazepam-treated animals Ro 15-1788 produced an increase of LCGU to values above control levels in those brain regions in which a decrement was seen upon acute diazepam administration to naive rats. These findings indicate an adaptation to and a possible development of physical dependence upon chronic drug treatment.

Animals↗
PubMed1985

Receptor binding and biological activity of bivalent enkephalins.

Two series of dimeric enkephalin analogues were assayed for opioid activity in two isolated smooth muscle preparations: the guinea pig ileum (GPI) and the mouse vas deferens (MVD). Dimers have the general structure: X-(CH2)n-X, where X is H-Tyr-D-Ala-Gly-Phe-Leu-NH-(n = 0, 2, 4, 6, 8, 10, 12), for the first series of dimeric pentapeptide enkephalins (DPEn), and H-Tyr-D-Ala-Gly-Phe-NH-(n = 2, 4, 6, 8, 12), for the series of dimeric tetrapeptide enkephalins (DTEn). Comparison of biological activities with binding affinities revealed that: (1) the DPE series with n = 2-8 showed increased potency in the MVD assay relative to monomeric [D-Ala2, Leu5]enkephalinamide (DALEA); (2) there was an associated increase affinity for the delta receptor of rat brain or neuroblastoma-glioma hybrid cells. (however, the relative potencies were higher in the MVD assay then predicted on the basis of binding affinities); (3) the DTE series also showed an increase in delta receptor affinities and MVD potencies relative to DALEA, for n = 2-12; (4) for the DTE series, the increase in MVD activities was less than that expected on the basis of delta binding affinity; (5) for both the DPE and DTE series, activities in the GPI assay and mu-receptor affinities were highly correlated: as the length of the methylene bridge increased from 2 to 12, there was a progressive loss of activity in both assays, with a similar pattern for DPE and DTE. Two selected dimers and their corresponding monomers were also assayed for antinociceptive activity in vivo: results were consistent with GPI and mu-binding but not with MVD and delta-binding. Two alkylamide analogs of penta- and tetrapeptide monomers, representing the monomer with the attached spacer of the most active dimers, were also assayed in biological and binding assays. Comparison of these compounds with the corresponding dimers suggest that the changes in activities and selectivities induced by dimerization are not a spurious effect of the presence of an akylamide derivative of the carboxy terminal of enkephalin but rather may represent a specific effect due to the bivalent nature of the ligands.

Analgesia↗
PubMed1985

Multiple states of opioid receptors may modulate adenylate cyclase in intact neuroblastoma X glioma hybrid cells.

Opioid receptor binding and opioid-mediated inhibition of cAMP accumulation were studied simultaneously in intact NG108-15 cells. The dose-response curves for the biological response were suggestive of positive cooperativity and systematically occurred at lower ligand concentrations than those for the binding of [3H] [D-Ala2, D-Leu5]enkephalin (DADLE), which were instead shallow and suggestive of a site heterogeneity or of a cooperative phenomenon. Computer modeling of the binding isotherms revealed that the data are best described assuming two binding sites with different affinities for the agonist; the mean ratio between the DADLE concentrations yielding half-maximal occupancy of the high affinity site and half-maximal response was 1.5, but it was 36 when the fractional occupancy of the sum of the two sites was considered. On examining several opioids, no direct correlation was found between high affinity site and biological response; however, several agonists displayed different affinities for the two sites, while the antagonist naloxone and the partial agonist diprenorphine bound to them with identical affinities. Furthermore, naloxone exhibited a good agreement between half-maximal receptor occupancy and Ki in blocking the agonist response. Thus, the binding heterogeneity detectable in intact cells is agonist-specific, and suggests rather that the sites are states of an identical receptor population. When [3H]diprenorphine was used to label the opioid receptors, the competition curves for DADLE were consistent with the existence of an additional, very low affinity state undetectable by direct binding with labeled agonist and, again, not discriminated by naloxone. Multiple affinity states of the opioid receptor in intact cells may reflect its interaction with the effector system in the plasma membrane.

Adenylyl Cyclases↗
PubMed1984

Sodium regulation of opioid agonist binding is potentiated by pertussis toxin.

Pretreatment of intact NG108-15 cells with pertussis toxin suppresses opioid inhibition of cyclic AMP accumulation mediated by the inhibitory guanine nucleotide-binding regulatory protein, Ni, which apparently also mediates the inhibitory nucleotide effects on opioid against binding. The toxin treatment had no effect on opioid agonist binding measured in NG108-15 cell membranes without sodium present. However, the toxin potentiated the inhibitory effect of sodium on agonist binding, leading to an agonist-specific reduction of opioid receptor affinity in the presence of sodium in the binding reaction. The potency of the stable GTP analog, GTP gamma S, to reduce agonist binding in the presence of sodium was little changed in membranes prepared from pertussis toxin-treated cells compared to control membranes, whereas the potency of the stable GDP analog, GDP beta S, was magnified. The data indicate that ADP-ribosylation of Ni by pertussis toxin potentiates sodium regulation of opioid agonist binding and that the communication between Ni and opioid receptors is not lost by the covalent modification of Ni.

Animals↗
PubMed1984

Receptor preference of dynorphin A fragments in the mouse vas deferens determined by different techniques.

The opioid receptor preference of dynorphin A fragments, particularly of dynorphin A-(1-8) (DYN 8), has been evaluated in the mouse vas deferens by means of cross-tolerance studies, by their sensitivity to naloxone antagonism and by the use of the irreversible narcotic antagonist beta-funaltrexamine. The tolerance studies revealed kappa receptor activity for the longer fragments and delta activity for the shorter fragments. DYN 8 displayed kappa as well as delta activity, whereas no interaction with mu receptors was observed. The naloxone sensitivity of dynorphin A and its fragments was low with the exception of DYN 8, that displayed an intermediate sensitivity. There was no indication that this intermediate value for DYN 8 was due to an interaction with mu receptors. This conclusion was strengthened in experiments using beta-funaltrexamine. The kappa and delta activity of DYN 8 does not explain the intermediate sensitivity to naloxone. It is proposed that DYN 8 may interact in the mouse vas deferens with a different opioid receptor than the classical mu, kappa- and delta-type.

Animals↗
PubMed1983

The molecular size of multiple opiate receptors.

The functional size of different types of opiate receptors was determined in situ by the radiation inactivation technique. The Mr of opiate binding sites was estimated to be about 90,000 daltons; the data revealed no significant differences between the molecular sizes of either mu-, delta- or kappa-sites in rat brain membranes. An identical molecular size was found also for the delta-sites in the neuroblastoma X glioma hybrid cell clone, NG 108-15.

Animals↗
PubMed1983

Pertussis toxin decreases opiate receptor binding and adenylate inhibition in a neuroblastoma x glioma hybrid cell line.

Neuroblastoma X glioma hybrid cells NG108-15 were treated with a toxin derived from Bordetella pertussis. As compared to control cells grown in the absence of toxin, the inhibitory effects of opioid agonists upon cAMP formation were dose-dependently impaired by a non-competitive mechanism. Radioligand binding studies revealed that opioid agonist binding was dramatically reduced in toxin-treated membranes when tested in the presence of Na+/Mg++/GMP-PNP. Further, the potencies of guanine nucleotides to decrease opioid agonist binding were differentially modulated. These studies may facilitate our understanding of the mechanisms responsible for acute and chronic opiate effects.

Adenylyl Cyclase Inhibitors↗
PubMed1983

Uncoupling of receptors is essential for opiate-induced desensitization (tolerance) in neuroblastoma x glioma hybrid cells NG 108-15.

Neuroblastoma x glioma hybrid cells NG 108-15 were chronically treated with opioid agonists and tested for their development of "tolerance" as evaluated by the loss of inhibitory activity of opioids upon cAMP-accumulation. Desensitization was dose-dependent and non-competitive and highly selective, since the activity of other, non-opioid inhibitory compounds, such as noradrenaline and carbachol, was not altered. Moreover, naloxone-induced "withdrawal signs" (revealed in the lack of change in cAMP) were lacking in preparations completely desensitized ("tolerant") towards opiate effects. These results reject the assumption of a common biochemical mechanism underlying both opiate tolerance and dependence and rather support the significance of an uncoupling of receptors from subsequent effector systems upon chronic opiate action.

Animals↗
PubMed1983

Multiple endorphins in neuronal hybrid cell lines.

Neuroblastoma x glioma hybrid cells (NG 108CC15) and tumors derived thereof were examined for dynorphin- and alpha-neoendorphin-like material. The techniques employed for analyses of opiate-like material were the isolated mouse vas deferens bioassay and gel chromatography and high pressure liquid chromatography in combination with radioimmunoassays. Dynorphin- and alpha-neoendorphin-like material was detected in both the hybrid cells and the corresponding tumors. Immunoreactive dynorphin and alpha-neoendorphin was also in NCB 20 hybrid cells and in tumors thereof assayed. In all samples investigated, the amounts of alpha-neoendorphin-like material was higher than that of dynorphin-like material. The results revealed considerable variability in the amount of dynorphin- and alpha-neoendorphin-activity between particular samples, suggesting the need for studies into the responsible mechanisms.

Animals↗
PubMed1983

A subclassification of multiple opiate receptors by means of selective tolerance development.

The chronic activation of opiate receptors results in the development of tolerance. On theoretical grounds, the appearance of cross-tolerance between different opioids should imply that these compounds exhibit an identical pattern on receptor activation. Tests with isolated tissue preparations, e.g. the guinea-pig ileum, made highly tolerant in vivo to the mu-agonist morphine proved that these were of almost unchanged sensitivity to different types of receptor agonists such as kappa-agonists. Thus, the ability to selectively induce tolerance on particular types of opiate receptors represents a reliable tool for the differentiation and characterization of multiple opiate receptors. Moreover, this technique facilitated the demonstration of subtypes of the already known opiate receptors.

Animals↗
PubMed1982

Opioid dependence and cross-dependence in the isolated guinea-pig ileum.

The development of opioid dependence and tolerance attributed to selective types of opiate receptors was studied in the isolated ileum of guinea pigs chronically exposed to specific opioids. These investigations were based on reports that in this preparation highly tolerant opiate receptors may coexist with opiate receptors of almost unchanged sensitivity. Thus, the ilea were set up in vitro and tested for tolerance and dependence. Apparently precipitation of the withdrawal contracture, indicating dependence, proved a more sensitive parameter than the phenomenon of tolerance. Maximal dependence was determined at rather low degrees of tolerance (5 to 10 fold). The intensity of the withdrawal contracture failed to increase as opiate tolerance did. Furthermore, the experiments failed to present evidence for the existence of selective dependence at specific opiate receptor types. These findings may suggest multiple adaptational mechanisms upon chronic activation of opiate receptors. One mechanism may be responsible for the development of dependence and a low degree of tolerance, whilst a further increase of tolerance may be associated with changes at the opiate binding site level.

Animals↗
PubMed1982

Opiate receptor binding studies in the mouse vas deferens exhibiting tolerance without dependence.

The apparent lack of dependence in highly opiate-tolerant isolated mouse vas deferens may conflict with unitary theories postulating a common biochemical mechanism for both phenomena. Therefore, attention focused on binding sites of the opiate receptors which in this tissue are located presynaptically. Binding studies conducted with homogenate of highly tolerant vasa deferentia revealed no significant different results as compared to naive tissues. Further studies examined the effect of guanine nucleotide on opiate receptor interaction. Apparently, mu-, delta- and kappa-opiate receptors in the mouse vas deferens proved resistant to the regulatory action of guanine nucleotide in naive and tolerant tissues. From these experiments, it is concluded that the binding characteristics of opiate receptors in the mouse vas deferens do not change with chronic activation. In addition, the lack of an effect of guanine nucleotide on opiate binding leads to the suggestion that binding sites are not coupled to adenylate cyclase in this tissue. Taken together, these findings draw the attention to the coupling mechanism of opiate receptors in the mouse vas deferens which may play a key role in the adaptational mechanisms following chronic opiate exposure.

Animals↗
PubMed1981

Are there subtypes (isoreceptors) of multiple opiate receptors in the mouse vas deferens?

The hypothesis of heterogeneity of opiate receptors was tested with the electrically stimulated mouse vas deferens. The experimental approach was based on the phenomenon of tolerance development of selective opiate receptors. It was found that opioids classified as mu-receptor agonists, e.g. sufentanyl and normorphine, do not necessarily exhibit cross-tolerance to each other. The same holds true for a variety of presumed kappa-receptor agonists. On the other hand, the delta-receptor agonists under investigation displayed complete cross-tolerance. These findings give rise to an interpretation which may postulate the existence of subtypes (isoreceptors) of the already known opiate receptors.

Animals↗