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Biomedical subjects

J Deckert

Publications and source records attributed to J Deckert.

At least 73 records · Page 4Linked to original sources

[Autoradiography determination of the GABA(A) receptor density in the brain of rats with portacaval shunt].

Quantitative autoradiography was used to assess the densities of gamma-aminobutyric acid (GABAA) receptors in the brains of rats with a portacaval end-to-side shunt (PCA). The shunt alone induced only mild encephalopathy with ataxia and decreased locomotion. Aggravation of the encephalopathy was achieved by gavage feeding of packed erythrocytes or by induction of severe hyperammonemia by intraperitoneal injection of urease. Gavage feeding of erythrocytes led to severe encephalopathy in about 50% of the animals with PCA. The combination of PCA and urease treatment caused severe encephalopathy in every animal. The serum ammonia concentration increased 5 times normal by PCA alone, 20 times normal by gavage feeding of erythrocytes and more than 30 times normal by urease-treatment of the PCA-animals. For autoradiography, coronal slices were cut at the level of the hippocampal formation and through the cerebellum. Radioligand binding was measured using as a ligand 3H-muscimol, a specific GABAA receptor agonist. The specific binding of 3H-muscimol was assessed densitometrically in several microregions of cerebral cortex, hippocampus and cerebellar cortex. No significant differences were observed between the magnitude of ligand binding to specific microregions of brains from normal animals, animals with PCA without overt encephalopathy and animals with severe encephalopathy induced by a combination of PCA and gavage feeding of erythrocytes or urease treatment.

Ammonia↗

Binding of [3H]inositoltrisphosphate and [3H]phorbol 12,13-dibutyrate in rat hippocampus following transient global ischemia: a quantitative autoradiographic study.

An in vitro quantitative autoradiographic binding study of the phosphatidylinositol system ligands [3H]inositol 1,4,5-trisphosphate (IP3) and [3H]phorbol 12,13-dibutyrate (PDBU) to rat brain slices was performed at 6, 12, 24, 28 and 72 h following a 20 min ischemic injury. PDBU binding showed a transient 20% decrease in the dentate gyrus and the CA3 the first 24 h as well as a 50% decrease in the CA1 at 72 h. A 50% decline in IP3 binding was seen in all regions at 6-12 h except the pyramidal cell layer of the CA1. This downregulation of calcium mobilizing intracellular receptors is probably a defence against ischemic neuronal cell death.

Animals↗

Protective effect of cyclohexyladenosine on adenosine A1-receptors, guanine nucleotide and forskolin binding sites following transient brain ischemia: a quantitative autoradiographic study.

The effects of an i.p. administration of cyclohexyladenosine (CHA) have been examined upon ischemic brain damage in gerbils. Ischemia was induced for 20 min by occlusion of both carotid arteries, and CHA was administered 5 min after recirculation at a dose of 2 mg/kg. Animals were sacrificed either 1, 3 or 6 days after ischemia and their brains were used for examination of cell morphology and quantitative autoradiography. In animals subject to ischemia, the deterioration of the laminar organization of the hippocampus was associated with a significant decrease in adenosine A1-receptors (labeled with [3H]CHA), G-protein (labeled with [3H]forskolin). The treatment with CHA considerably improved the morphological preservation of cells in the CA1 region of the hippocampus and prevented the reduction in the specific binding of all radioligands. Adenosine, its analogues and other substances modulating adenosine receptors may thus provide new therapeutic approaches to the treatment of ischemia-induced brain injury.

Adenosine↗

Delayed c-fos proto-oncogene expression in the rat hippocampus induced by transient global cerebral ischemia: an in situ hybridization study.

The relative levels of c-fos mRNA in individual neurons of the hippocampal formation of rats is dramatically increased following 20 min of cerebral ischemia induced by 4-vessel occlusion. After 24 h of recirculation, a number of scattered neurons in the dentate hilus became hybridization positive. This effect appeared to peak between 24 and 48 h. A few neurons in the pyramidal cell layer of CA1 expressed c-fos as early as 24 h, but the most intense labeling in this region was seen at 72 h of recirculation. These results correlate well with the known distribution of delayed ischemic necrosis in the brain.

Animals↗

Autoradiographic analysis of GABA-benzodiazepine receptors in an animal model of acute hepatic encephalopathy.

To complement analogous studies using conventional ligand-membrane binding assays, the densities of gamma-aminobutyric acid and benzodiazepine receptors in the brain have been assessed using an autoradiographic technique in an animal model of hepatic encephalopathy. Hepatic encephalopathy due to fulminant hepatic failure was induced in rabbits by the intravenous injection of galactosamine. The specific binding of three radiolabeled ligands was assessed densitometrically in several microregions of cerebral cortex, hippocampus and cerebellum. [3H]Muscimol was used to assess gamma-aminobutyric acid receptor density and [3H]flunitrazepam or [3H]Ro 15-1788 was used to assess benzodiazepine receptor density. No significant differences were observed between the magnitude of binding of the three ligands to each of the microregions of brain from control rabbits and rabbits in Stage III or IV hepatic encephalopathy. These findings suggest that the behavioral expression of hepatic encephalopathy in the model studied is not dependent upon an increase in the number of gamma-aminobutyric acid or benzodiazepine receptors, but do not conflict with the hypothesis that gamma-aminobutyric acid-ergic tone is increased in hepatic encephalopathy.

Acute Disease↗

Potent convulsant actions of the adenosine receptor antagonist, xanthine amine congener (XAC).

The convulsant properties of xanthine amine congener (XAC, 8-(4-(2-aminoethyl)-aminocarboxylmethyloxy)phenyl-1,3-dipropylxant hine) are compared to those of caffeine. Male Swiss albino mice were infused with convulsants through a lateral tail vein. Convulsion thresholds (i.e. the amount of convulsants required to elicit convulsions) of 39.8 +/- 2.0 mg/kg (n = 10) and 109.8 +/- 2.3 mg/kg (n = 10) were calculated for XAC and caffeine respectively. Pretreatment of animals with the adenosine receptor agonists 2-chloroadenosine, N6-cyclohexyladenosine or 5'-N-ethylcarboxamido-adenosine (1 mg/kg, i.p., 20 minutes prior to infusion) significantly decreased the seizure threshold of both XAC and caffeine. The adenosine uptake blockers, 6-nitrobenzylthioinosine or dipyridamole (0.25 mg/kg, i.p., 20 minutes prior to infusion) did not significantly affect the seizure threshold to either XAC or caffeine. The benzodiazepine agonist diazepam (5 mg/kg, i.p., 20 minutes prior to infusion) significantly increased the seizure threshold to both XAC (p less than 0.05) and caffeine (p less than 0.01), whereas the benzodiazepine antagonist Ro 15-1788 (10 mg/kg, i.p., 20 minutes prior to infusion) significantly increased the seizure threshold to caffeine (p less than 0.01), but not XAC. The results suggest that actions at benzodiazepine receptors may be a tenable hypothesis to explain the convulsant actions of caffeine, but not those of XAC.

Animals↗

Electroconvulsive shock (ECS) and the adenosine neuromodulatory system: effect of single and repeated ECS on the adenosine A1 and A2 receptors, adenylate cyclase, and the adenosine uptake site.

The effect of a single electroconvulsive shock (ECS) (30 min and 24 h after treatment) and repeated ECS (10 once-daily) on the adenosine neuromodulatory system was investigated in rat cerebral cortex, cerebellum, hippocampus, and striatum. The present study examined the adenosine A1 receptor using N6-[3H]cyclohexyladenosine ([3H]CHA), the A2 receptor using 5'-N-[3H]ethylcarboxyamidoadenosine ([ 3H]NECA), adenylate cyclase using [3H]forskolin, and the adenosine uptake site using [3H]nitrobenzylthioinosine ([3H]NBI). At 30 min after a single ECS, the Bmax of the [3H]NBI binding in striatum was increased by 20%, which is in good agreement with the well-known postictal adenosine release. The Bmax of [3H]forskolin binding in striatum and cerebellum was increased by 60 and 20%, respectively. In contrast to earlier reported changes following chemically induced seizures, [3H]CHA binding was not altered postictally. At 24 h after a single ECS, there were no changes for any ligand in any brain region. Following repeated ECS, there was a 20% increase of [3H]CHA binding sites in cerebral cortex, which lasted for at least 14 days after the last ECS. [3H]Forskolin binding in hippocampus and striatum was 20% lowered 24 h after 10 once-daily ECS but had already returned to control levels 48 h after the last treatment. Evidence is provided that the upregulated adenosine A1 receptors are coupled to guanine nucleotide binding proteins and, furthermore, that this upregulation is not paralleled by an increase in adenylate cyclase activity as labeled by [3H]forskolin.

Adenosine↗

Regional ontogenetic profile of central and peripheral benzodiazepine receptors in the guinea pig brain.

The ontogenetic development of 'central' and 'peripheral' benzodiazepine (BZ) receptors was investigated in 3 different regions of the guinea pig brain. A marked difference in the developmental profile of these two types of binding sites was observed. The density of central-type receptors gradually increased during embryonic life (especially in cortex and cerebellum), without major changes after birth, while the reverse was true for peripheral-type binding sites. These data contrast with those previously reported in rat, suggesting species differences.

Aging↗

Ontogeny of adenosine uptake sites in guinea pig brain: differential profile of [3H]nitrobenzylthioinosine and [3H]dipyridamole binding sites.

The ontogenetic profile of adenosine uptake sites was investigated in guinea pig cerebral cortex and cerebellum using as ligand probes the uptake inhibitors, [3H]nitrobenzylthioinosine ([3H]NBI) and [3H]dipyridamole ([3H]DPR). In cerebral cortex [3H]NBI binding was highest at E50 and decreased subsequently until P28 while in cerebellum after a first peak at E50 and a subsequent decline it increased again until P28. [3H]DPR binding increased by 25% from E40 to P28 in cerebral cortex while in cerebellum hardly any binding could be detected before E50 and it afterwards increased by more than 250% until P28. Scatchard analysis demonstrated that [3H]NBI labeled approximately as many sites as [3H]DPR in cerebral cortex at E44 while at P28 [3H]DPR labeled more than double as many sites. Accordingly, NBI was more potent in displacing [3H]DPR binding at E44 than at P28. These findings suggest that part of the [3H]DPR binding sites, i.e. the NBI-insensitive one develops later than [3H]NBI binding sites during ontogeny in guinea pig cerebral cortex and cerebellum.

Aging↗

Evidence for pre- and postsynaptic localization of adenosine A1 receptors in the CA1 region of rat hippocampus: a quantitative autoradiographic study.

The cellular localization of adenosine A1 receptors in the CA1 region of the rat hippocampus was investigated using radioligand binding and lesioning methods in a quantitative autoradiographic study. Kainic acid injection reduced [3H]cyclohexyladenosine binding in the CA3 region by 40% and in the CA1 region by 30%. Transient cerebral ischemia reduced [3H]cyclohexyladenosine binding in the CA1 region by 50%, while no change was measured in the CA3 region. These findings suggest that adenosine A1 receptors are located presynaptically on axon terminals of CA3 pyramidal cells and postsynaptically on the dendrites of CA1 pyramidal cells in the CA1 region of rat hippocampus.

Adenosine↗

Autoradiographic localization of mouse brain adenosine receptors with an antagonist ([3H]xanthine amine congener) ligand probe.

A [3H]xanthine amine congener (XAC), a potent adenosine receptor antagonist, binds in a saturable and reversible fashion to high affinity binding sites in mouse brain (Bmax = 323 +/- 17 fmol/mg protein, Kd = 1.4 +/- 0.4 nM). Adenosine receptor agonists and antagonists are more potent than adenosine uptake inhibitors in displacing the binding of [3H]xanthine amine congener ([3H]XAC). The anatomical distribution of [3H]XAC binding sites is consistent with its being a ligand probe for adenosine receptors. High binding site densities were observed in the hippocampus (stratum oriens and radiatum, molecular layer), superior colliculus (superficial gray), cerebellum (molecular layer), cerebral cortex and substantia nigra. The availability of a high affinity antagonist radioligand probe like [3H]XAC for adenosine receptors allows the comparative quantitative autoradiographic analysis of agonist and antagonist binding to adenosine receptors, e.g. under varying in vitro incubation conditions (presence and absence of guanine nucleotides and cations).

Animals↗

The effect of acute and chronic electroconvulsive shock on [3H]phorbol-dibutyrate binding to rat brain membranes.

The present study investigated the effect of single and repeated electroconvulsive shock (ECS) on proteinkinase C in rat cerebral cortex, cerebellum, hippocampus and striatum using [3H]Phorbol-12,13-butyrate binding. In the postictal period and 24 hr after a single ECS there was no alteration in any brain region. Twenty four hr after 10 once-daily ECS there was a significant decrease the number of binding sites in cerebral cortex (30%) and in cerebellum (20%) without a change in the affinity constant. These findings are discussed with regard to earlier reports on phosphoinositide turnover following chemically and electrically induced seizures.

Animals↗

Adenosine uptake site heterogeneity in the mammalian CNS? Uptake inhibitors as probes and potential neuropharmaceuticals.

Inhibitors of adenosine uptake or transport have been used clinically for some time in certain cardiovascular diseases. More recently, some of them have also been investigated for possible clinical use in combination with antimetabolites based on the observed heterogeneity of nucleoside transport in mammalian tumor cells. Such a heterogeneity of adenosine uptake and uptake sites has now also been suggested in the mammalian CNS. The aim of this article is, therefore, to review the present status of our knowledge of adenosine uptake in the mammalian CNS, compare it with our far more advanced knowledge of nucleoside transport in other mammalian cells and suggest direction of future research. The possible implications for the development of adenosine uptake inhibitors as adenosinergic neuropharmaceuticals will be discussed based on our knowledge of the physiological function of adenosine in the CNS.

Adenosine↗

Adenosine uptake sites in brain: regional distribution of putative subtypes in relationship to adenosine A1-receptors.

Adenosine uptake sites have been characterized and localized in guinea pig and pointer dog brain by in vitro autoradiography, using as probes 3H-nitrobenzylthioinosine (3H-NBI) and the recently available 3H-dipyridamole (3H-DPR). In guinea pig brain and, to a lesser extent, in pointer dog brain, 3H-DPR was found to label more high-affinity binding sites than 3H-NBI and NBI inhibited 3H-DPR binding having pseudo-Hill coefficients smaller than 0.5. 3H-DPR and 3H-NBI labeled brain structures with different intensities in guinea pig brain, as was revealed by quantitative analysis. While the intensity of 3H-DPR binding varied about 4-fold in neuron-containing structures, 8-fold differences were observed for 3H-NBI binding with phylo- and ontogenetically older brain areas such as hypothalamus and various brain stem structures showing relatively higher densities. These findings raise the interesting possibility of adenosine uptake site heterogeneity (NBI-sensitive and insensitive) in guinea pig brain, complementing the well-established adenosine receptor heterogeneity (A1 and A2). As adenosine's neurodepressant effects are believed to be mainly mediated by adenosine A1-receptors, these were localized using 3H-cyclohexyl-adenosine (3H-CHA) as a ligand probe. In guinea pig brain, the highest receptor densities were seen in hippocampus and claustrum, while only relatively low levels were found in hypothalamus and various brain stem structures. As was previously described for rat brain, major discrepancies in the regional distribution of adenosine A1-receptors and adenosine uptake sites, as labeled by 3H-NBI, were seen in guinea pig brain. These discrepancies were only partly abolished (e.g., in cerebellum) by the use of 3H-DPR as an additional ligand probe for adenosine uptake sites. Adenosine uptake site heterogeneity, therefore, probably does not explain the previously described discrepancies in rodent brain between the distribution of adenosine A1-receptors and uptake sites. Because of the low affinity of 3H-DPR for adenosine uptake sites in rat and mouse brain, these species could not be investigated with this new radioligand probe. In pointer dog brain, as compared to guinea pig brain, a more similar distribution pattern of adenosine A1-receptors and adenosine uptake sites in the brain structures investigated (e.g., hippocampus) could be observed. The situation in guinea pig brain can, therefore, not be universalized to other species.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine↗