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S Rossner

Publications and source records attributed to S Rossner.

53 records · Page 3Linked to original sources
PubMed1995

Differential changes in cholinergic markers from selected brain regions after specific immunolesion of the rat cholinergic basal forebrain system.

The aim of this study was to characterize the effects of cortical cholinergic denervation on cholinergic parameters in the cerebral cortex and basal forebrain using a novel immunotoxin (conjugate of the monoclonal antibody 192IgG against the low-affinity nerve growth factor receptor armed with cytotoxin saporin) to efficiently and selectively lesion cholinergic neurons in rat basal forebrain. Seven days following an intracerebroventricular injection of the cholinergic immunotoxin 192IgG-saporin the binding levels of nicotinic and M1- and M2-muscarinic acetylcholine receptors (mAChR), high-affinity choline uptake sites, as well as the m1-m4 mAChR mRNA were determined in coronal brain sections by both receptor autoradiography and in situ hybridization, and quantified by image analysis. Hemicholinium-3 binding to high-affinity choline uptake sites was decreased by up to 45% in all cortical regions and in the hippocampus after a single injection of the immunotoxin compared to controls. In contrast, M1-mAChR sites were increased over the corresponding control value in the anterior parts of cingulate, frontal, and piriform cortex by about 20%, in the hindlimb/forelimb areas (18%), in the parietal cortex (35%), in the occipital cortex area 2 (17%), as well as in the temporal cortex (25%) following immunolesion. M2-mAChR levels were found to be significantly increased in the posterior part of the parietal cortex area 1 (by about 22%) and in the occipital cortex area 2 (20%) only. With respect to laminar cortical localization, M2-mAChRs and choline uptake sites were altered in all cortical layers, whereas M1-mAChRs were preferentially affected in the upper cortical layers by the immunolesion. The increase in M1-mAChR binding in the temporal and occipital cortex as a consequence of the immunolesion was complemented by an increase in the amount of m1 and m3 mAChR mRNA of about 20% in these regions. The elevated levels of M2-mAChR sites in the occipital and temporal cortex following immunolesion were accompanied by an increase in the m4 (by 25%) but not m2 mAChR mRNA. There was no effect of the immunolesion on the m1-m4 mAChR mRNA in frontal cortical regions. in the basal forebrain, however, immunolesioning caused about a 40% decrease in the level of m2 mAChR mRNA in the medial and lateral septum as well as in the vertical and horizontal limb of the diagonal band, whereas M1- and M2-mAChR binding and the levels of m1, m3, and m4 mAChR mRNA were not affected by the immunolesion in any of the basal forebrain nuclei studied.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

192IGG-saporin-induced selective lesion of cholinergic basal forebrain system: neurochemical effects on cholinergic neurotransmission in rat cerebral cortex and hippocampus.

A novel cholinergic immunotoxin (conjugate of the monoclonal antibody 192IgG against the low-affinity nerve growth factor receptor with the cytotoxin saporin) producing selective lesions of cholinergic neurons in rat basal forebrain was applied to study its effect on hippocampal and cerebral cortical cholinergic neurotransmission. Intracerebroventricular injection of 4 micrograms 192IgG-saporin conjugate resulted in a selective loss of cholinergic cells in the basal forebrain nuclei 1 week after application, which was accompanied by decreased activities of choline acetyltransferase and by reduced high-affinity uptake of [3H]choline into cholinergic nerve terminals in the cerebral cortex and hippocampus, as well as by a significant activation of micro- and to a lesser extent of astroglial cells in the hippocampus, but hardly in the cerebral cortex.. The K(+)-stimulated release of [3H]acetylcholine from cortical and hippocampal slices of immunolesioned rats was found to be markedly decreased 1 week after injection. Cholinergic immunolesion led to enhanced cortical M1-muscarinic acetylcholine receptor numbers, but did not alter muscarinic receptor sensitivity as measured by carbachol-stimulated inositol phosphate production or phorbol ester binding to membrane-bound protein kinase C. In the hippocampal formation differential enhancements in binding levels of M1-muscarinic cholinergic receptor sites in the CA1 region and in the dentate gyrus were observed, whereas the nicotinic and M2-muscarinic receptor subtype are seemingly not affected by the immunotoxin in either of the subfields studied. Cholinergic immunolesioning did not result in any alterations in the hybridization signals for m1 through m4 muscarinic acetylcholine receptor mRNA in any region or layer of the hippocampus. The data suggest that (i) the novel cholinergic immunotoxin 192IgG-saporin is an appropriate tool to mimic cholinergic hypofunction in the hippocampal formation and cerebral cortex, and (ii) selective and specific immunolesion of cholinergic cells in medial septal nuclei differentially affects cholinergic receptors in particular hippocampal subfields.

Animals↗
PubMed1994

Ibotenic acid lesion of nucleus basalis magnocellularis differentially affects cholinergic, glutamatergic and GABAergic markers in cortical rat brain regions.

The present study was undertaken to study the effect of reduced cortical cholinergic activity on gamma-aminobutyric acid (GABA)ergic and glutamatergic mechanisms in cholinoceptive cortical target regions which are assumed to play an important role for realizing cognitive functions. The densities of cortical muscarinic cholinergic receptor subtypes and corresponding receptor genes m1 through m4, N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) and kainate glutamate receptor subtypes as well as GABAA and benzodiazepine receptors were measured in rats 1 week after unilateral ibotenic acid lesion of the nucleus basalis magnocellularis (Nbm) applying quantitative receptor autoradiography and in situ hybridization. Ibotenic acid lesion resulted in a striking loss of acetylcholinesterase (AChE) staining in the lesioned Nbm which is associated with a 60% decrease in AChE staining and a 30% reduction in [3H]hemicholinium-3 binding in frontal and parietal cortical regions as well fore- and hindlimb areas ipsilateral to the lesion, being more prominent in the more rostral cortical regions. M1-muscarinic cholinergic receptor binding was not changed in any of the cortical regions studied 1 week after lesion. M2-muscarinic receptor binding levels are slightly increased in the parietal cortex only. The lesion-induced increase in parietal cortical M2-muscarinic receptor binding is complemented by an increase in the hybridization signal for the corresponding m4-mRNA transcript. In cortical regions displaying a reduced activity of AChE and decreased levels of high-affinity choline uptake sites due to forebrain cholinergic lesion, NMDA receptor binding was markedly reduced in comparison to the unlesioned brain side whereas AMPA and kainate binding has been significantly increased in these regions. Muscimol binding to GABAA receptors was increased in the rostral portions of frontal and parietal cortices as compared with the unlesioned brain side. Binding levels of benzodiazepine receptors were not affected by the lesion in any of the cortical regions studied. The differential changes in glutamate and GABA receptor subtypes following lesion might be regarded as the consequence of a cortical reorganization compensating for the reduced cholinergic presynaptic input. The data further suggest that presynaptic cortical cholinergic deficits might affect both glutamatergic and GABAergic functions with different intensity and different directions.

Acetylcholine↗
PubMed1994

Differential expression of immediate early genes in distinct layers of rat cerebral cortex after selective immunolesion of the forebrain cholinergic system.

The aim of this study was to show whether reduction or loss of cortical cholinergic activity results in any particular change in the expression of the proto-oncogenes c-fos and/or c-jun. To produce cortical cholinergic hypofunction, the monoclonal antibody, 192IgG, to the low-affinity nerve growth factor receptor p75NGFR coupled to a cytotoxin, saporin, was used as an efficient and selective immunotoxin for cholinergic neurons in rat basal forebrain. Brain sections of adult rats that had received an intracerebroventricular injection of 4 micrograms of the 192IgG-saporin were subjected to in situ hybridization using oligonucleotides to detect c-fos and c-jun mRNA. Autoradiographs obtained were evaluated by quantitative image analysis. Seven days following injection of the immunotoxin there was a dramatic loss in acetylcholinesterase staining in frontal, parietal, piriform, temporal, and occipital cortices, hippocampus, and olfactory bulb, but not in the striatum and cerebellum. In situ hybridization revealed a considerable increase in the level of c-fos mRNA in the lateral septum following the cholinergic lesion, whereas in the medial septum both c-fos and c-jun mRNA were elevated. Immunolesioning led to a distinct and specific increase in the level of c-jun but not c-fos mRNA in the parietal and occipital cortex that was restricted to cortical layer IV. These data suggest that reduced cortical cholinergic activity differentially regulates expression of c-fos/c-jun genes in distinct cortical regions of the rat brain.

Acetylcholinesterase↗
PubMed1994

Development of laminar expression of the m2 muscarinic cholinergic receptor gene in rat visual cortex and the effect of monocular visual deprivation.

The postnatal development of laminar pattern of the m2 muscarinic acetylcholine receptor subtype mRNA in the visual cortex of both normally raised and monocularly deprived rats (one eyelid sutured at the age of 11 days) was studied using in situ hybridization histochemistry and computer-assisted image analysis. In normally raised rats, on birth, the m2 transcript was found to be more concentrated in the superficial zones of the cortex. This laminar pattern alters to a more homogeneous distribution of the label throughout the cortex already detectable on day 7. From day 10 onwards a bimodal laminar pattern gradually develops with increased mRNA levels in layer IV and upper layer VI. From postnatal day 21 onwards the hybridization peak in layer VI decreases as compared to the peak level in layer IV resulting in an adult distribution with highest labeling in layer IV, low labeling in layer I to III and moderate labeling in layers V and VI. Monocular deprivation results in decreased m2 mRNA levels in visual cortical layers IV-VI in both deprived and non-deprived cortices already detectable at the age of 18 days and persisting up to the age of 21 days; but this effect disappears following further deprivation until adulthood. The data suggest that the changes in m2 receptor level from a more homogeneous distribution to a bimodal pattern during postnatal development seem to be related to synaptogenesis and final tuning of connectional pattern within the rat visual cortex.

Animals↗
PubMed1994

Receptor function in cortical rat brain regions after lesion of nucleus basalis.

The present study was undertaken to study the interaction of cholinergic and glutamatergic mechanisms in cholinoceptive cortical target regions which is assumed to play an important role for realizing cognitive functions. The densities of cortical muscarinic cholinergic receptor subtypes and corresponding receptor genes m1 through m4, as well as NMDA, AMPA and kainate glutamate receptor subtypes were measured in rats one week after unilateral mechanical lesion of the anterior part of the nucleus basalis magnocellularis (NbM) applying quantitative receptor autoradiography and in situ hybridization. The studies revealed that in cortical regions displaying a low amount of acetylcholinesterase activity due to forebrain cholinergic lesion, NMDA receptor binding was markedly reduced in comparison to the unlesioned side, whereas AMPA and kainate binding has been significantly increased in these regions. M1-muscarinic cholinergic receptor binding was not changed in any of the cortical regions studied, whereas M2-receptor densities are slightly reduced in frontal and parietal cortices following lesion. These alterations in cortical M2-muscarinic receptor binding are complemented by corresponding changes in the m2- and m4-mRNA transcripts. The comparison of binding profiles through selected cortical regions of both lesioned and normal brain side revealed that lesion of the NbM affects NMDA receptors in all cortical layers of the lesioned side, whereas AMPA receptors are affected preferentially in the upper and kainate receptors preferentially in the middle and deeper cortical layers. The differential changes in glutamate receptor subtypes following lesion might be regarded as the consequence of a cortical reorganization compensating for the reduced cholinergic presynaptic input. The data further suggest that presynaptic cortical cholinergic deficits might affect glutamatergic functions with different intensity and different directions.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Muscarinic acetylcholine receptor subtypes in rat visual cortex--a comparative study using quantitative receptor autoradiography and in situ hybridization.

The laminar pattern of M1- and M2-muscarinic acetylcholine receptors (mAChR) in rat visual cortex has been compared with the distribution of the corresponding m1, m2, m3 and m4 receptor genes using both quantitative receptor autoradiography and in situ hybridization histochemistry. The laminar distribution of 3H-pirenzepine binding to M1-mAChRs in rat visual cortex shows a bimodal pattern with higher binding levels in upper layer III and deeper layer VI. In contrast, highest binding of 3H-AF-DX384 to M2-mAChRs was observed in upper layer IV (100%) and upper layer VI (about 80% of highest binding). The m1 receptor mRNA is almost homogeneously distributed throughout the visual cortex, whereas the m2mAChr mRNA predominates in layer IV with lower levels in layers I and V. The highest amounts of m3mAChR mRNA in rat visual cortex were observed in layer II, while the distribution of m4mAChR transcripts shows a bimodal pattern with peaks in layers III and upper layer VI. The distinct laminar pattern of mRNA muscarinic receptor subtypes in rat visual cortex suggest specific roles of the muscarinic receptor in visual function.

Animals↗
PubMed1993

Laminar expression of m1-, m3- and m4-muscarinic cholinergic receptor genes in the developing rat visual cortex using in situ hybridization histochemistry. Effect of monocular visual deprivation.

The postnatal development of laminar pattern of m1-, m3- and m4-mRNA-muscarinic acetylcholine receptor subtypes in the visual cortex of both normally raised and monocularly deprived rats (one eyelid sutured at the age of 11 days) was studied using in situ hybridization histochemistry and computer-assisted image analysis. From birth until day 15 the level of m1-receptor transcript in layer II/III increases markedly as compared to deeper layers. From day 15 up to day 18 a transient bimodal pattern develops with peaks in layers II/III and VI. Already on day 35 a more homogeneous distribution of m1-receptor mRNA level is detectable persisting until adulthood. In contrast, the m3-receptor mRNA shows already at birth a bimodal distribution with peaks in layers II/III and VI. Further development until adulthood results in transient changes in the ratio of the mRNA levels in these layers. In the adult visual cortex a similar laminar pattern as at birth is observed. From day 1 up to day 10 a relative increase in the mRNA level of the m4-receptor in layers II to IV is observed. From day 10 until day 15 a bimodal distribution of receptor mRNA develops with peaks in layers III and VI which is similar to the adult stage. However, between days 18 and 35 a shift in the laminar receptor mRNA distribution occurs resulting in peaks in layers IV and VI. The labeling of the m5-receptor transcript in rat visual cortex was very weak and did not show any alteration with age. Unilateral eyelid closure from postnatal day 11 resulted in transient changes in the laminar distribution of m3- and m4-receptor mRNA between postnatal days 18 and 25, whereas the development of the laminar pattern of the m1-receptor mRNA was not affected regardless of the length of visual deprivation. The distinct laminar developmental pattern of mRNA muscarinic receptor subtypes in rat visual cortex suggests specific roles of the muscarinic receptor subtypes during the first weeks of postnatal maturation of visual function.

Aging↗
PubMed1993

Differential laminar expression of AMPA receptor genes in the developing rat visual cortex using in situ hybridization histochemistry. Effect of visual deprivation.

The postnatal development of laminar pattern of AMPA receptor mRNA subtypes GluR-A through to GluR-D (flip variants) in the visual cortex of both normally raised and monocularly deprived rats (one eyelid sutured at the age of 11 days) was studied using in situ hybridization histochemistry and computer-assisted image analysis. The AMPA receptors GluR-A through to GluR-D transcripts exhibit a differential laminar expression pattern in the developing rat visual cortex. At birth the levels of GluR-A transcripts are lower by about 50% in each visual cortical layer as compared to the adult values. In contrast, GluR-B to GluR-D mRNAs are expressed in all cases at higher levels at birth than in the adult brain. Unilateral eyelid closure at postnatal day 11 for several periods of time resulted in both transient and permanent changes in the laminar development of GluR-A through to GluR-C transcripts but hardly affected the GluR-D mRNA subtype. The distinct laminar developmental pattern of AMPA receptor mRNAs in rat visual cortex as well as the differential effects of visual deprivation suggest specific roles of AMPA receptor subtypes during the early postnatal maturation of visual function.

Animals↗
PubMed1991

Evaluation of a new magnetic resonance imaging method for quantitating adipose tissue areas.

We have developed a new magnetic resonance imaging (MRI) protocol to quantitate intraabdominal and subcutaneous fat and have validated it by comparing measurements of fat areas by MRI with those obtained by computed tomography (CT) in 11 asymptomatic volunteers who all had a single CT and MRI image taken at the level of the umbilicus. The new MRI protocol was based on a water-fat separation method by which the slice selection routines excite water and fat protons in different positions along the slice select direction. This method performed more reliably than earlier methods based on phase differences between water and fat signals. Fat areas measured by MRI exceeded those measured by CT by 8-22 percent, and fat areas and ratios obtained by MRI correlated well with CT (r = 0.98 for areas and, for ratios, r = 0.81). The ratio of intraabdominal/subcutaneous fat measured by MRI in seven males was significantly greater than that in four females. We also compared the new method with a previously published inversion recovery (IR) method in seven additional volunteers. Agreement between the two methods was excellent, and the major differences were technical: the IR protocol produced images that may require custom image processing programs when obtained on some scanners. Comparability of the two methods provides further reassurance of the validity of both. MRI presents an attractive opportunity for directly measuring intraabdominal fat in order to correlate this with metabolic parameters and to visualize changes during weight loss.

Abdomen↗
PubMedDate not supplied

Cerebrospinal fluid cholinesterases--markers for loss of cholinergic basal forebrain neurons?

The present study was conducted to test the hypothesis that cholinergic basal forebrain neurons are a major source of cerebrospinal fluid (CSF) cholinesterases. To address this question enzyme activities of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in both CSF and parietal cortex were assayed following selective lesion of basal forebrain cholinergic neurons by a single intracerebroventricular application of the cholinergic immunotoxin 192IgG-saporin. Cholinergic immunolesions led to a dramatic decrease in total AChE activity in parietal cortex, which was due to the specific loss of the G4 molecular form while the activity of the G1 form was increased as compared to nonlesioned animals. In contrast, the total enzyme activity of BChE and its molecular forms were not affected by cholinergic lesion in both parietal cortex and CSF. The data suggest, that cholinergic basal forebrain neurons are seemingly not a major source of cholinesterases in the CSF, and do not provide any evidence for using CSF cholinesterases as a diagnostic marker of basal forebrain cholinergic cell loss in humans.

Acetylcholine↗
PubMedDate not supplied

Glucose metabolism in cholinoceptive cortical rat brain regions after basal forebrain cholinergic lesion.

To address the question whether the changes in cortical glucose metabolism observed in patients with Alzheimer's disease are interrelated with, or consequences of, basal forebrain cholinergic cell loss, an experimental approach was employed to produce cortical cholinergic dysfunction in rat brain by administration of the cholinergic immunotoxin 192IgG-saporin. [14C]D-glucose utilization in brain homogenates, D-glucose-displaceable [3H]cytochalasin B binding to glucose transporters (GLUT). Northern and Western analyses, as well as in vivo [14C]2-deoxyglucose autoradiography were used to quantify the regional glucose metabolism. Basal forebrain cholinergic lesion resulted in transient increases in glucose transporter binding in cortical regions displaying reduced acetylcholinesterase activity, already detectable seven days after lesion with peak values around 30 days post lesion. Western analysis revealed that the changes in total glucose transporter binding are mainly due to changes in the GLUT3 subtype only, while the levels of GLUT1 and GLUT3 mRNA (Northern analysis) were not affected by cholinergic lesion. Both immunocytochemistry and in situ hybridization demonstrated preferential localizations of GLUT1 on brain capillaries and GLUT3 on neurons, respectively. A lesion-induced transient decrease in [14C]D-glucose utilization seven days post lesion was detected in the lesion site, whereas cholinoceptive cortical regions were not affected. In vivo [14C]deoxyglucose uptake was transiently increased in cholinoceptive cortical regions and in the lesion site being highest between three to seven days after lesion. The cholinergic lesion-induced transient up-regulation of cortical glucose transporters and deoxyglucose uptake reflects an increased glucose demand in regions depleted by acetylcholine suggesting functional links between cortical cholinergic activity and glucose metabolism in cholinoceptive target regions.

Animals↗
PubMedDate not supplied

Muscarinic acetylcholine receptors activate the acetylcholinesterase gene promoter.

The acetylcholinesterase (AChE) gene promoter contains several overlapping binding sites for Sp1 and Egr-1 transcription factors. Cotransfection experiments and promoter assays showed that Egr-1 can potently activate transcription from the human AChE promoter. Muscarinic acetylcholine receptors (mAChR) rapidly activate, via protein kinase C-mediated signaling, expression of the Egr-1 gene, leading to dramatically increased nuclear concentrations of Egr-1 protein, and to increased binding of Egr-1 to specific DNA recognition sequences. These mAChR-induced increases are followed by increased transcription from the human AChE promoter. In vivo studies with intraventricular infusions of the cholinergic immunotoxin 192 IgG saporin showed more than 80% decrease of AChE activity in cholinergic target areas of the hippocampus and brain cortex. The results are compatible with a combination of decreased AChE activity in degenerating subcortical cholinergic projections, and additional decreases in postsynaptic AChE gene expression. Together our data show that mAChR can activate transcription from the AChE promoter via increased synthesis of Egr-1. The results suggest a feedback mechanism by which the AChE gene is activated by cholinergic neurotransmission, possibly leading to increased formation of AChE protein and accelerated degradation of acetylcholine at cholinergic synapses. This possibility suggests testing of cholinomimetic compounds currently in development for the treatment of Alzheimer's disease for their potential ability to increase AChE gene expression.

Acetylcholinesterase↗