Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 159
Filter
1.
BMC Cardiovasc Disord ; 23(1): 566, 2023 11 18.
Article in English | MEDLINE | ID: mdl-37980508

ABSTRACT

BACKGROUND: Induction of chondrogenesis is associated with progressive atherosclerosis. Deficiency of the ADCYAP1 gene encoding pituitary adenylate cyclase-activating peptide (PACAP) aggravates atherosclerosis in ApoE deficient (ApoE-/-) mice. PACAP signaling regulates chondrogenesis and osteogenesis during cartilage and bone development. Therefore, this study aimed to decipher whether PACAP signaling is related to atherogenesis-related chondrogenesis in the ApoE-/- mouse model of atherosclerosis and under the influence of a high-fat diet. METHODS: For this purpose, PACAP-/-/ApoE-/-, PAC1-/-/ApoE-/-, and ApoE-/- mice, as well as wildtype (WT) mice, were studied under standard chow (SC) or cholesterol-enriched diet (CED) for 20 weeks. The amount of cartilage matrix in atherosclerotic lesions of the brachiocephalic trunk (BT) with maximal lumen stenosis was monitored by alcian blue and collagen II staining on deparaffinized cross sections. The chondrogenic RUNX family transcription factor 2 (RUNX2), macrophages [(MΦ), Iba1+], and smooth muscle cells (SMC, sm-α-actin) were immunohistochemically analyzed and quantified. RESULTS: ApoE-/- mice fed either SC or CED revealed an increase of alcian blue-positive areas within the media compared to WT mice. PAC1-/-/ApoE-/- mice under CED showed a reduction in the alcian blue-positive plaque area in the BT compared to ApoE-/- mice. In contrast, PACAP deficiency in ApoE-/- mice did not affect the chondrogenic signature under either diet. CONCLUSIONS: Our data show that PAC1 deficiency reduces chondrogenesis in atherosclerotic plaques exclusively under conditions of CED-induced hypercholesterolemia. We conclude that CED-related chondrogenesis occurs in atherosclerotic plaques via transdifferentiation of SMCs and MΦ, partly depending on PACAP signaling through PAC1. Thus, PAC1 antagonists or PACAP agonists may offer therapeutic potential against pathological chondrogenesis in atherosclerotic lesions generated under hypercholesterolemic conditions, especially in familial hypercholesterolemia. This discovery opens therapeutic perspectives to be used in the treatment against the progression of atherosclerosis.


Subject(s)
Atherosclerosis , Plaque, Atherosclerotic , Animals , Mice , Plaque, Atherosclerotic/pathology , Pituitary Adenylate Cyclase-Activating Polypeptide/genetics , Chondrogenesis/physiology , Alcian Blue , Atherosclerosis/genetics , Atherosclerosis/pathology , Cholesterol , Diet, High-Fat , Apolipoproteins E/genetics , Mice, Knockout , Mice, Inbred C57BL
2.
Virology ; 511: 82-94, 2017 11.
Article in English | MEDLINE | ID: mdl-28841446

ABSTRACT

The Rabies lyssavirus glycoprotein (RABV-G) is largely responsible for the neuroinvasiveness of the virus and the induction of antiviral immune responses. To study the effects of RABV-G we compared the G of the attenuated RABV variant SPBN with that of the pathogenic DOG4 strain. Infection via the olfactory route caused 100% mortality in mice with both virus variants. Of note, with the attenuated SPBN, progression of the disease was accelerated, microglia response less pronounced and IL-6 expression higher than in the presence of RABV-G from the pathogenic DOG4. However, while virus spread was less extensive, viral gene expression in individual neurons was actually higher in SPBN-infected brains without causing apoptosis of infected neurons. These differences between the two variants were not observed in infected neuronal cultures indicating that the effects of RABV-G on virus spread and viral gene expression depend on factors only present in the intact brain.


Subject(s)
Antigens, Viral/genetics , Antigens, Viral/metabolism , Brain/virology , Glycoproteins/genetics , Glycoproteins/metabolism , Neurons/virology , Rabies virus/isolation & purification , Rabies/virology , Viral Envelope Proteins/genetics , Viral Envelope Proteins/metabolism , Viral Load , Animals , Apoptosis , Disease Models, Animal , Gene Expression Profiling , Genes, Viral , Mice , Survival Analysis , Virulence
3.
Int Immunopharmacol ; 29(1): 158-65, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26033492

ABSTRACT

Specialized epithelial cells in the respiratory tract such as solitary chemosensory cells and brush cells sense the luminal content and initiate protective reflexes in response to the detection of potentially harmful substances. The majority of these cells are cholinergic and utilize the canonical taste signal transduction cascade to detect "bitter" substances such as bacterial quorum sensing molecules. Utilizing two different mouse strains reporting expression of choline acetyltransferase (ChAT), the synthesizing enzyme of acetylcholine (ACh), we detected cholinergic cells in the submucosal glands of the murine larynx and trachea. These cells were localized in the ciliated glandular ducts and were neither found in the collecting ducts nor in alveolar or tubular segments of the glands. ChAT expression in tracheal gland ducts was confirmed by in situ hybridization. The cholinergic duct cells expressed the brush cell marker proteins, villin and cytokeratin-18, and were immunoreactive for components of the taste signal transduction cascade (Gα-gustducin, transient receptor potential melastatin-like subtype 5 channel = TRPM5, phospholipase C(ß2)), but not for carbonic anhydrase IV. Furthermore, these cells expressed the bitter taste receptor Tas2r131, as demonstrated utilizing an appropriate reporter mouse strain. Our study identified a previously unrecognized presumptive chemosensory cell type in the duct of the airway submucosal glands that likely utilizes ACh for paracrine signaling. We propose that these cells participate in infection-sensing mechanisms and initiate responses assisting bacterial clearance from the lower airways.


Subject(s)
Acetylcholine/metabolism , Chemoreceptor Cells/metabolism , Epithelial Cells/metabolism , Larynx/cytology , Trachea/cytology , Animals , Choline O-Acetyltransferase/genetics , Choline O-Acetyltransferase/metabolism , Gene Expression Regulation, Enzymologic , Green Fluorescent Proteins , Mice , Mice, Inbred C57BL , Mice, Transgenic
4.
Neuroscience ; 298: 302-17, 2015 Jul 09.
Article in English | MEDLINE | ID: mdl-25892702

ABSTRACT

L-3,4-Dihydroxyphenylalanine (L-DOPA) is the therapeutic gold standard in Parkinson's disease. However, long-term treatment is complicated by the induction of debilitating abnormal involuntary movements termed L-DOPA-induced dyskinesias (LIDs). Until today the underlying mechanisms of LID pathogenesis are not fully understood. The aim of this study was to reveal new factors, which may be involved in the induction of LID. We have focused on the expression of striatal tyrosine hydroxylase-positive (TH+) neurons, which are capable of producing either L-DOPA or dopamine (DA) in target areas of ventral midbrain DAergic neurons. To address this issue, a daily L-DOPA dose was administered over the course of 15 days to mice with unilateral 6-hydroxydopamine-induced lesions of the medial forebrain bundle and LIDs were evaluated. Remarkably, the number of striatal TH+ neurons strongly correlated with both induction and severity of LID as well as ΔFosB expression as an established molecular marker for LID. Furthermore, dyskinetic mice showed a marked augmentation of serotonergic fiber innervation in the striatum, enabling the decarboxylation of L-DOPA to DA. Axial, limb and orolingual dyskinesias were predominantly associated with TH+ neurons in the lateral striatum, whereas medially located TH+ neurons triggered locomotive rotations. In contrast, identified accumbal and cortical TH+ cells did not contribute to the generation of LID. Thus, striatal TH+ cells and serotonergic terminals may cooperatively synthesize DA and subsequently contribute to supraphysiological synaptic DA concentrations, an accepted cause in LID pathogenesis.


Subject(s)
Corpus Striatum/pathology , Dyskinesia, Drug-Induced/pathology , Functional Laterality/physiology , Neurons/metabolism , Tyrosine 3-Monooxygenase/metabolism , Amphetamine/pharmacology , Animals , Antiparkinson Agents/adverse effects , Disease Models, Animal , Dyskinesia, Drug-Induced/etiology , Levodopa/adverse effects , Male , Medial Forebrain Bundle/drug effects , Medial Forebrain Bundle/injuries , Mice , Mice, Inbred C57BL , Oxidopamine/toxicity , Parkinson Disease/drug therapy , Parkinson Disease/etiology , Phosphopyruvate Hydratase/metabolism , Proto-Oncogene Proteins c-fos/metabolism , Serotonin Plasma Membrane Transport Proteins/metabolism , Statistics, Nonparametric
5.
Diabetologia ; 56(5): 1047-56, 2013 May.
Article in English | MEDLINE | ID: mdl-23404442

ABSTRACT

AIMS/HYPOTHESIS: Imaging of beta cell mass (BCM) is a major challenge in diabetes research. The vesicular monoamine transporter 2 (VMAT2) is abundantly expressed in human beta cells. Radiolabelled analogues of tetrabenazine (TBZ; a low-molecular-weight, cell-permeant VMAT2-selective ligand) have been employed for pancreatic islet imaging in humans. Since reports on TBZ-based VMAT2 imaging in rodent pancreas have been fraught with confusion, we compared VMAT2 gene expression patterns in the mouse, rat, pig and human pancreas, to identify appropriate animal models with which to further validate and optimise TBZ imaging in humans. METHODS: We used a panel of highly sensitive VMAT2 antibodies developed against equivalently antigenic regions of the transporter from each species in combination with immunostaining for insulin and species-specific in situ hybridisation probes. Individual pancreatic islets were obtained by laser-capture microdissection and subjected to analysis of mRNA expression of VMAT2. RESULTS: The VMAT2 protein was not expressed in beta cells in the adult pancreas of common mouse or rat laboratory strains, in contrast to its expression in beta cells (but not other pancreatic endocrine cell types) in the pancreas of pigs and humans. VMAT2- and tyrosine hydroxylase co-positive (catecholaminergic) innervation was less abundant in humans than in rodents. VMAT2-positive mast cells were identified in the pancreas of all species. CONCLUSIONS/INTERPRETATION: Primates and pigs are suitable models for TBZ imaging of beta cells. Rodents, because of a complete lack of VMAT2 expression in the endocrine pancreas, are a 'null' model for assessing interference with BCM measurements by VMAT2-positive mast cells and sympathetic innervation in the pancreas.


Subject(s)
Insulin-Secreting Cells/metabolism , Pancreas/metabolism , Vesicular Monoamine Transport Proteins/metabolism , Aged , Animals , Female , Gene Expression Regulation , Humans , Insulin-Secreting Cells/cytology , Insulin-Secreting Cells/immunology , Ligands , Male , Mast Cells/cytology , Mast Cells/immunology , Mast Cells/metabolism , Mice , Middle Aged , Nerve Endings/metabolism , Pancreas/cytology , Pancreas/immunology , Pancreas/innervation , Radioligand Assay , Rats , Species Specificity , Sus scrofa , Sympathetic Nervous System/cytology , Sympathetic Nervous System/metabolism , Tetrabenazine/analogs & derivatives , Tetrabenazine/metabolism , Vesicular Monoamine Transport Proteins/genetics
6.
Histochem Cell Biol ; 137(4): 483-97, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22261922

ABSTRACT

The luminal composition of the auditory tube influences its function. The mechanisms involved in the monitoring are currently not known. For the lower respiratory epithelium, such a sentinel role is carried out by cholinergic brush cells. Here, using two different mouse strains expressing eGFP under the control of the promoter of choline acetyltransferase (ChAT), we show the presence of solitary cholinergic villin-positive brush cells also in the mouse auditory tube epithelium. They express the vesicular acetylcholine (ACh) transporter and proteins of the taste transduction pathway such as α-gustducin, phospholipase C beta 2 (PLC(ß2)) and transient receptor potential cation channel subfamily M member 5 (TRPM5). Immunoreactivity for TRPM5 and PLCß2 was found regularly, whereas α-gustducin was absent in approximately 15% of the brush cells. Messenger RNA for the umami taste receptors (TasR), Tas1R1 and 3, and for the bitter receptors, Tas2R105 and Tas2R108, involved in perception of cycloheximide and denatonium were detected in the auditory tube. Using a transgenic mouse that expresses eGFP under the promotor of the nicotinic ACh receptor α3-subunit, we identified cholinoceptive nerve fibers that establish direct contacts to brush cells in the auditory tube. A subpopulation of these fibers displayed also CGRP immunoreactivity. Collectively, we show for the first time the presence of brush cells in the auditory tube. These cells are equipped with all proteins essential for sensing the composition of the luminal microenvironment and for communication of the changes to the CNS via attached sensory nerve fibers.


Subject(s)
Chemoreceptor Cells/cytology , Cholinergic Neurons/cytology , Eustachian Tube/cytology , Animals , Mice , Mice, Inbred C57BL , Mice, Transgenic , Tongue/cytology
7.
Anesthesiol Res Pract ; 2011: 591851, 2011.
Article in English | MEDLINE | ID: mdl-21716736

ABSTRACT

Background. Nerve stimulation can facilitate correct needle placement in peripheral regional anesthesia. The aim of this study was to determine whether the high threshold current is associated with reduced nerve injury due to fewer needle-nerve contacts compared with low current. Methods. In anaesthetized pigs, thirty-two nerves of the brachial plexus underwent needle placement at low (0.2 mA) or high current (1.0 mA). The occurrence of needle-nerve contact was recorded. After 48 hours, the nerves were analyzed for occurrence of histological changes. Nerve injury was scored ranging from 0 (no injury) to 4 (severe injury). Results. The frequency of needle-nerve contact was 94% at low compared to 6% at high current. The score was significantly higher at low (median [interquartile range] 2.0 [1.0-2.0]) compared to high current (0.0 [0.0-1.0] P = .001). Conclusions. Inflammatory responses were directly related to needle-nerve contacts. Hence, posttraumatic inflammation may be diminished using higher current for nerve localization.

8.
Regul Pept ; 165(1): 36-44, 2010 Nov 30.
Article in English | MEDLINE | ID: mdl-20005907

ABSTRACT

Immunoreactivity for both processed and unprocessed forms of chromogranin A (CGA) was examined, using an antibody recognizing the WE14 epitope, among terminal fields and cell bodies of anatomically defined GABAergic, glutamatergic, cholinergic, catecholaminergic, and peptidergic cell groups in the rodent central nervous system. CGA is ubiquitous within neuronal cell bodies, with no obvious anatomical or chemically-coded subdivision of the nervous system in which CGA is not expressed in most neurons. CGA expression is essentially absent from catecholaminergic terminal fields in the CNS, suggesting a relative paucity of large dense-core vesicles in CNS compared to peripheral catecholaminergic neurons. Extensive synaptic co-localization with classical transmitter markers is not observed even in areas such as amygdala, where CGA fibers are numerous, suggesting preferential segregation of CGA to peptidergic terminals in CNS. Localization of CGA in dendrites in some areas of CNS may indicate its involvement in regulation of dendritic release mechanisms. Finally, the ubiquitous presence of CGA in neuronal cell somata, especially pronounced in GABAergic neurons, suggests a second non-secretory vesicle-associated function for CGA in CNS. We propose that CGA may function in the CNS as a prohormone and granulogenic factor in some terminal fields, but also possesses as-yet unknown unique cellular functions within neuronal somata and dendrites.


Subject(s)
Central Nervous System/metabolism , Chromogranin A/metabolism , Neurons/metabolism , Animals , Immunohistochemistry , In Vitro Techniques , Male , Mice
9.
Neuroscience ; 156(2): 310-8, 2008 Oct 02.
Article in English | MEDLINE | ID: mdl-18722510

ABSTRACT

Classic neurotransmitter phenotypes are generally predetermined and develop as a consequence of target-independent lineage decisions. A unique mode of target-dependent phenotype instruction is the acquisition of the cholinergic phenotype in the peripheral sympathetic nervous system. A body of work suggests that the sweat gland plays an important role to determine the cholinergic phenotype at this target site. A key issue is whether neurons destined to innervate the sweat glands express cholinergic markers before or only after their terminals make target contact. We employed cholinergic-specific over-expression of the vesicular acetylcholine transporter (VAChT) in transgenic mice to overcome sensitivity limits in the detection of initial cholinergic sweat gland innervation. We found that VAChT immunoreactive nerve terminals were present around the sweat gland anlage already from the earliest postnatal stages on, coincident selectively at this sympathetic target with tyrosine hydroxylase-positive fibers. Our results provide a new mechanistic model for sympathetic neuron-target interaction during development, with initial selection by the target of pioneering nerve terminals expressing a cholinergic phenotype, and subsequent stabilization of this phenotype during development.


Subject(s)
Acetylcholine/metabolism , Neurons/metabolism , Norepinephrine/metabolism , Phenotype , Sweat Glands/innervation , Sympathetic Nervous System/cytology , Age Factors , Animals , Animals, Newborn , Choline O-Acetyltransferase/genetics , Embryo, Mammalian , Gene Expression Regulation, Developmental/physiology , Green Fluorescent Proteins/biosynthesis , Green Fluorescent Proteins/genetics , Mice , Mice, Transgenic , Sympathetic Nervous System/growth & development , Tyrosine 3-Monooxygenase/metabolism , Vesicular Acetylcholine Transport Proteins/genetics , Vesicular Acetylcholine Transport Proteins/metabolism
10.
Dev Biol (Basel) ; 131: 73-81, 2008.
Article in English | MEDLINE | ID: mdl-18634468

ABSTRACT

Rabies virus (RABV) infection is characterized by the rapid neuronal spread of RABV into the CNS before a protective immune response is raised. Therefore, a typical feature of RABV infection is the paucity of inflammatory reactions in the brain. Here we examined whether the induction of immunosuppressive neuropeptides, in particular CGRP, may contribute to the ability of RABV to evade immune responses. RABV infection of mice caused a strong induction of calcitonin gene-related peptide (CGRP) in neurons and fibres in the neocortex as well as in the dentate gyrus and CA1 region of the hippocampus although RABV did not infect neurons in which CGRP expression was upregulated. Neuropeptide Y (NPY) or vasoactive intestinal peptide (VIP) expressing neurons also were not infected by RABV. In contrast, somatostatin neurons were infected by RABV. There was evidence for an RABV-induced increase of VIP and somatostatin but not of NPY. To test how CGRP expression is related to TNFalpha-induced enhancement of CNS innate and adaptive immunity during RABV infection, we used recombinant RABVs that contained either an active (SPBN-TNFalpha(+)) or an inactive (SPBN-TNFalpha(-)) TNFalpha gene. As compared to SPBN-TNFalpha(-), infection with SPBN-TNFalpha(+) attenuated the induction of CGRP but simultaneously enhanced induction of the invariant chain of MHC II, microglial activation and T cell infiltration. In conclusion, distinct neuropeptidergic neurons in the brain are remarkably spared from RABV infection suggesting a pivotal role of neuropeptides during CNS virus infection. Given the inhibitory effect of CGRP on antigen presentation, we propose that the strong RABV-induced upregulation of CGRP in the brain may contribute to the mechanism by which RABV escapes immune detection. Targeting the expression of neuropeptides, in particular CGRP, that are induced during RABV infection may open a new avenue for therapeutic intervention in human rabies.


Subject(s)
Brain/metabolism , Central Nervous System Viral Diseases/immunology , Neuropeptides/metabolism , Rabies virus/immunology , Rabies/immunology , Animals , Brain/pathology , Brain/virology , Central Nervous System Viral Diseases/metabolism , Central Nervous System Viral Diseases/pathology , Female , Mice , Neuropeptides/genetics , Rabies/metabolism , Rabies/pathology , Up-Regulation
11.
J Periodontal Res ; 42(3): 259-66, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17451546

ABSTRACT

BACKGROUND AND OBJECTIVE: The aim of this study was to identify the expression of cyclooxygenase-1, cyclooxygenase-2, cyclooxygenase-3, and microsomal prostaglandin E synthase-1 in young and elderly subjects. MATERIAL AND METHODS: Periodontally healthy subjects were divided into young (18-30 years, n = 7) and elderly (46-77 years, n = 7). A gingival biopsy was taken at baseline. After experimental gingivitis, clinical examination was repeated and a second biopsy was taken. The expression of cyclooxygenase-1, cyclooxygenase-2, cyclooxygenase-3, and microsomal prostaglandin E synthase-1 was analyzed by means of immunohistochemistry. RESULTS: In both healthy age groups, cyclooxygenase-1 and microsomal prostaglandin E synthase-1 were expressed in epithelial cells, endothelial cells and fibroblast-like connective tissue cells. Cyclooxygenase-1 was found in Langerhans' cells of the epithelium. Cyclooxygenase-2 expression was observed in cells exhibiting the morphology of epithelial mitosis cells, and the expression of cyclooxygenase-2 in periodontally healthy elderly subjects was significantly lower (p < or = 0.05). Following experimental gingivitis, cyclooxygenase-1 and microsomal prostaglandin E synthase-1 expression did not change. However, the expression of cyclooxygenase-2 was significantly increased in both age groups (p < or = 0.05). Cyclooxygenase-3 was not detected in any group investigated. CONCLUSION: Cyclooxygenase-1 and microsomal prostaglandin E synthase-1 were expressed constitutively in gingival tissue, and expression was unaffected by age or inflammation states. In contrast, the expression of cyclooxygenase-2 was weaker in elderly subjects. In the course of experimental gingivitis, cyclooxygenase-2 was induced in both age groups.


Subject(s)
Gingiva/enzymology , Gingivitis/enzymology , Prostaglandins/biosynthesis , Adolescent , Adult , Age Factors , Aged , Biopsy/methods , Cyclooxygenase 1/analysis , Cyclooxygenase 2/analysis , Epidemiologic Methods , Female , Gingiva/cytology , Gingivitis/etiology , Humans , Intramolecular Oxidoreductases/analysis , Male , Middle Aged , Prostaglandin-E Synthases , Prostaglandin-Endoperoxide Synthases/analysis
12.
Gut ; 56(5): 637-44, 2007 May.
Article in English | MEDLINE | ID: mdl-17135306

ABSTRACT

BACKGROUND: Patients with a multiple endocrine neoplasia type 1 (MEN1)-associated Zollinger-Ellison syndrome (ZES) show multifocal duodenal gastrinomas and precursor lesions. AIMS: To test these lesions for loss of heterozygosity (LOH) of the MEN1 gene locus on chromosome 11q13, and to investigate whether the MEN1-related endocrine cell changes also involved somatostatin cells. MATERIAL AND METHODS: Tissue specimens from six patients with MEN1 and ZES were analysed by immunohistochemistry and immunofluorescence. LOH analysis was performed by fluorescence in situ hybridisation (FISH), using probes containing the MEN1 gene locus and the centromere 11 (C11) region. For simultaneous analysis of hormones and allelic deletions, a combined FISH/immunofluorescence protocol was established. RESULTS: 28 of a total of 33 duodenal neuroendocrine tumours (NETs) were gastrin-producing tumours; 13/28 (46.4%) revealed LOH on 11q13 and/or C11. Five of the NETs were somatostatin-expressing tumours, two revealing LOH. Allelic loss was detected in tumours as small as 300 microm (gastrin) and 400 microm (somatostatin) in diameter. The gastrin-producing tumours showed different deletion/retention patterns. Hyperplastic somatostatin cell lesions, similar to those of the gastrin cells, were present in all patients. The hyperplastic lesions of both cell lines consistently retained both 11q13 alleles. CONCLUSIONS: Allelic deletion of the MEN1 gene may reflect a pivotal event in the development of multifocal gastrin and somatostatin cell neoplasms in the duodenum of patients with MEN1. The observation of distinct deletion patterns in small synchronous tumours supports the concept that each gastrin-producing tumour in an individual MEN1 patient arises from an independent cell clone.


Subject(s)
Duodenal Neoplasms/genetics , Gastrinoma/genetics , Loss of Heterozygosity , Multiple Endocrine Neoplasia Type 1/genetics , Proto-Oncogene Proteins/genetics , Adult , Chromosomes, Human, Pair 11/genetics , Duodenal Neoplasms/pathology , Female , Gastrinoma/pathology , Humans , Hyperplasia/genetics , In Situ Hybridization, Fluorescence , Male , Middle Aged , Multiple Endocrine Neoplasia Type 1/pathology , Precancerous Conditions/genetics , Precancerous Conditions/pathology , Zollinger-Ellison Syndrome/genetics , Zollinger-Ellison Syndrome/pathology
13.
Naunyn Schmiedebergs Arch Pharmacol ; 365(1): 22-8, 2002 Jan.
Article in English | MEDLINE | ID: mdl-11862330

ABSTRACT

The anti-parkinsonian drug selegiline is a monoamine oxidase B (MAO-B) inhibitor and a potential neuroprotective agent which facilitates dopaminergic transmission. Its metabolites (-)-amphetamine and (-)-metamphetamine might contribute to the pharmacological effects as they are also able to increase dopaminergic transmission and in addition might lead to behavioural sensitization after repeated administration. We investigated the effects of acute and repeated treatment with a high dose of selegiline on dopamine overflow in the striatum as well as on behaviour and on tyrosine hydroxylase (TH) mRNA levels in midbrain. Two experiments were performed. In the first one, rats were implanted with microdialysis probes into the striatum and received daily injections of selegiline (10 mg/kg, i.p.) for 1 or 8 days or a single dose of saline. In vivo microdialysis was carried out on days 1, 8 or 17 (after withdrawal of 9 days) to measure dopamine overflow. Motility was measured at the same time. In the second experiment, rats were injected daily with selegiline (10 mg/kg, i.p.) or saline over a time period of 6 weeks or only once before the brains were processed for in situ hybridization with a (35)S-radiolabelled probe for TH. Repeated treatment led to higher levels in motility scores than acute administration after administration of the same dose, indicating behavioural sensitization, which was still manifest after an interruption of 9 days in the supply of selegiline. In contrast, acute administration of selegiline increased dopamine levels to a similar degree as the same dose after subchronic treatment, with or without interruption of 9 days. The dopamine metabolite DOPAC was reduced by more than 50% after acute administration of selegiline and even more so on day 8 by the same dose, after repeated administration. The basal concentrations of dopamine (before challenge with selegiline) were not altered by the repeated administration, whereas the basal concentrations of DOPAC were decreased by more than 80% by the repeated administration of selegiline, suggesting a decrease in dopamine turnover. Acute administration did not have any influence on TH mRNA levels, whereas chronic treatment significantly reduced TH mRNA levels in substantia nigra and ventral tegmental area. In conclusion, repeated administration of selegiline leads to behavioural sensitization independent of altered dopamine levels. In addition, it leads to a decrease, probably due to a down-regulation, of dopamine turnover and tyrosine hydroxylase.


Subject(s)
Antiparkinson Agents/administration & dosage , Corpus Striatum/drug effects , Dopamine/metabolism , Motor Activity/drug effects , RNA, Messenger/metabolism , Selegiline/administration & dosage , Tyrosine 3-Monooxygenase/metabolism , Animals , Corpus Striatum/enzymology , Drug Administration Schedule , In Situ Hybridization , Injections, Intraperitoneal , Male , Motor Activity/physiology , Rats , Rats, Wistar , Tyrosine 3-Monooxygenase/genetics
14.
J Virol ; 75(22): 10800-7, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11602721

ABSTRACT

The pathogenicity of individual rabies virus strains appears to correlate inversely with the extent of apoptotic cell death they induce and with the expression of rabies virus glycoprotein, a major inducer of an antiviral immune response. To determine whether the induction of apoptosis by rabies virus contributes to a decreased pathogenicity by stimulating antiviral immunity, we have analyzed these parameters in tissue cultures and in mice infected with a recombinant rabies virus construct that expresses the proapoptotic protein cytochrome c. The extent of apoptosis was strongly increased in primary neuron cultures infected with the recombinant virus carrying the active cytochrome c gene [SPBN-Cyto c(+)], compared with cells infected with the recombinant virus containing the inactive cytochrome c gene [SPBN-Cyto c(-)]. Mortality in mice infected intranasally with SPBN-Cyto c(+) was substantially lower than in SPBN-Cyto c(-)-infected mice. Furthermore, virus-neutralizing antibody (VNA) titers were significantly higher in mice immunized with SPBN-Cyto c(+) at the same dose. The VNA titers induced by these recombinant viruses paralleled their protective activities against a lethal rabies virus challenge infection, with SPBN-Cyto c(+) revealing an effective dose 20 times lower than that of SPBN-Cyto c(-). The strong increase in immunogenicity, coupled with the marked reduction in pathogenicity, identifies the SPBN-Cyto c(+) construct as a candidate for a live rabies virus vaccine.


Subject(s)
Cytochrome c Group/biosynthesis , Rabies virus/enzymology , Animals , Antibodies, Viral/blood , Female , Immunization , Mice , Mice, Inbred C3H , Nucleocapsid/analysis , Nucleocapsid Proteins , Phenotype , Rabies Vaccines/immunology , Rabies virus/genetics , Rabies virus/immunology , Recombination, Genetic
15.
J Immunol ; 167(6): 3470-7, 2001 Sep 15.
Article in English | MEDLINE | ID: mdl-11544340

ABSTRACT

We have recently demonstrated that increased blood-CNS barrier permeability and CNS inflammation in a conventional mouse model of experimental allergic encephalomyelitis are dependent upon the production of peroxynitrite (ONOO(-)), a product of the free radicals NO* and superoxide (O2*(-)). To determine whether this is a reflection of the physiological contribution of ONOO(-) to an immune response against a neurotropic pathogen, we have assessed the effects on adult rats acutely infected with Borna disease virus (BDV) of administration of uric acid (UA), an inhibitor of select chemical reactions associated with ONOO(-). The pathogenesis of acute Borna disease in immunocompetent adult rats results from the immune response to the neurotropic BDV, rather than the direct effects of BDV infection of neurons. An important stage in the BDV-specific neuroimmune response is the invasion of inflammatory cells into the CNS. UA treatment inhibited the onset of clinical disease, and prevented the elevated blood-brain barrier permeability as well as CNS inflammation seen in control-treated BDV-infected rats. The replication and spread of BDV in the CNS were unchanged by the administration of UA, and only minimal effects on the immune response to BDV Ags were observed. These results indicate that the CNS inflammatory response to neurotropic virus infection is likely to be dependent upon the activity of ONOO(-) or its products on the blood-brain barrier.


Subject(s)
Blood-Brain Barrier/drug effects , Borna Disease/immunology , Borna disease virus/immunology , Brain/immunology , Chemotaxis, Leukocyte/physiology , Encephalitis, Viral/immunology , Free Radical Scavengers/therapeutic use , Neuroprotective Agents/therapeutic use , Peroxynitrous Acid/physiology , Tyrosine/analogs & derivatives , Uric Acid/therapeutic use , Acute Disease , Animals , Antigens, Viral/immunology , Borna Disease/pathology , Borna Disease/virology , Borna disease virus/physiology , Brain/metabolism , Brain/pathology , Brain/virology , Brain Chemistry/drug effects , Chemotaxis, Leukocyte/drug effects , Encephalitis, Viral/pathology , Encephalitis, Viral/virology , Female , Free Radical Scavengers/pharmacology , Free Radicals , Gene Expression Profiling , Immunocompetence , Inflammation , Lymphocyte Count , Nerve Tissue Proteins/analysis , Neurons/enzymology , Neuroprotective Agents/pharmacology , Nitric Oxide Synthase/analysis , Nitric Oxide Synthase Type II , Oxidation-Reduction , Polymerase Chain Reaction , Rats , Rats, Inbred Lew , T-Lymphocyte Subsets/drug effects , Tyrosine/analysis , Uric Acid/pharmacology , Virus Replication/drug effects
16.
Neuroscience ; 104(3): 633-42, 2001.
Article in English | MEDLINE | ID: mdl-11440797

ABSTRACT

The cholinergic phenotype requires the expression of the vesicular acetylcholine transporter and choline acetyltransferase proteins. Both genes are encoded at one chromosomal location called the cholinergic gene locus. We have identified by in situ hybridization histochemistry distinct patterns of transcription from the cholinergic gene locus in the subdivisions of the rat cholinergic nervous system. The vesicular acetylcholine transporter and choline acetyltransferase are co-expressed in cholinergic neurons at all developmental stages in all major types of cholinergic neurons. The relative levels of vesicular acetylcholine transporter and choline acetyltransferase transcripts, however, change substantially during development in the CNS. They also differ dramatically in distinct subdivisions of the mature cholinergic nervous system, with vesicular acetylcholine transporter mRNA expressed at high levels relative to choline acetyltransferase mRNA in the peripheral nervous system, but at equivalent levels in the CNS. Expression of the R-exon, the presumptive first non-coding exon common to both the vesicular acetylcholine transporter and choline acetyltransferase, was not detectable at any developmental stage in any of the cholinergic neuronal subtypes in the rat nervous system. Thus, in contrast to less complex metazoan organisms, production of the vesicular acetylcholine transporter and choline acetyltransferase via a common differentially spliced transcript does not seem to occur to a significant extent in the rat. We suggest that separate transcriptional start sites within the cholinergic gene locus control vesicular acetylcholine transporter and choline acetyltransferase transcription, while additional elements are responsible for the specific transcriptional control of the entire locus in cholinergic versus non-cholinergic neurons. Independent transcription of the vesicular acetylcholine transporter and choline acetyltransferase genes provides a mechanism for regulating the relative expression of these two proteins to fine-tune acetylcholine quantal size in different types of cholinergic neurons, both centrally and peripherally.


Subject(s)
Acetylcholine/genetics , Gene Expression Regulation, Developmental/physiology , Locus Control Region/genetics , Membrane Transport Proteins , Nervous System/embryology , Nervous System/growth & development , Neurons/metabolism , Transcription, Genetic/physiology , Vesicular Transport Proteins , Acetylcholine/biosynthesis , Aging/genetics , Animals , Carrier Proteins/biosynthesis , Carrier Proteins/genetics , Choline O-Acetyltransferase/biosynthesis , Choline O-Acetyltransferase/genetics , Exons/genetics , Female , Fetus , Nervous System/metabolism , Neurons/cytology , Pregnancy , RNA, Messenger/metabolism , Rats , Rats, Sprague-Dawley , Vesicular Acetylcholine Transport Proteins
17.
Naunyn Schmiedebergs Arch Pharmacol ; 363(3): 288-94, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11284443

ABSTRACT

After repeated administration of cocaine at intervals, sensitization phenomena can be observed, so that its behavioural effects are enhanced. Since this phenomenon is long-lasting, it was of interest to study which persistent alterations in the activity of dopaminergic neurones or of endogenous opioid systems downstream of dopaminergic synapses in the basal ganglia are involved in the sensitization. Cocaine (10 mg/kg i.p.) was administered to rats on days 1, 3, 5 and 7 and saline on days 2, 4 and 6 ("repeated cocaine"), or saline was injected on days 1-6 and cocaine on day 7 ("acute cocaine"), or saline was injected on days 1-7 ("saline group"). The "repeated cocaine" schedule led to a significant sensitization to the locomotor activation produced by cocaine on day 7 or on day 17, 10 days after the end of sensitization protocol. Microdialysis in the nucleus accumbens which was performed after administration of cocaine (10 mg/kg i.p.) on day 7, or after an administration of the same dose 10 days after the last administration of cocaine, respectively, revealed significant acute increases of extracellular dopamine to about 200% of basal values. These increases were similar in "acute cocaine" and in "repeated cocaine" animals both after 7 days and after 17 days. For in situ hybridization studies, rats were sacrificed on day 7, 4.5 h after the last cocaine or saline administration. The mRNA for tyrosine hydroxylase (TH) in substantia nigra + ventral tegmental area was significantly elevated to about 140% of saline controls both in the "repeated cocaine" and the "acute cocaine" group as compared with the "saline group". In contrast, there were no differences between the three groups in the mRNAs of preprodynorphin or preproenkephalin levels measured in the nucleus accumbens (core and shell). These results suggest that sensitization phenomena to cocaine are not necessarily connected with alterations in the dopaminergic activity in the mesolimbic system or in the transcription of precursors of endogenous opioid peptides which are located downstream of the dopaminergic synapses.


Subject(s)
Behavior, Animal/drug effects , Cocaine/pharmacology , Dynorphins/genetics , Enkephalins/genetics , Protein Precursors/genetics , RNA, Messenger/metabolism , Tyrosine Transaminase/genetics , Animals , Dopamine/metabolism , Dopamine/physiology , Dynorphins/biosynthesis , Enkephalins/biosynthesis , In Situ Hybridization , Locomotion/drug effects , Male , Microdialysis , Neurons/drug effects , Neurons/metabolism , Neurons/physiology , Nucleus Accumbens/drug effects , Nucleus Accumbens/enzymology , Nucleus Accumbens/metabolism , Protein Precursors/biosynthesis , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Rats , Rats, Wistar , Tyrosine Transaminase/biosynthesis
18.
Neuropsychopharmacology ; 24(5): 531-44, 2001 May.
Article in English | MEDLINE | ID: mdl-11282253

ABSTRACT

Antidepressants produce various immunomodulatory effects, as well as an attenuation of the behavioral responses to immune challenges, such as lipopolysaccharide (LPS). To explore further the effects of antidepressants on neuroimmune interactions, rats were treated daily with either fluoxetine (Prozac) or saline for 5 weeks, and various behavioral, neuroendocrine, and immune functions were measured following administration of either LPS or saline. Chronic fluoxetine treatment significantly attenuated the anorexia and body weight loss, as well as the depletion of CRH-41 from the median eminence and the elevation in serum corticosterone levels induced by LPS. Chronic treatment with imipramine also attenuated LPS-induced adrenocortical activation. In rats and in mice, which normally display a biphasic body temperature response to LPS (initial hypothermia followed by hyperthermia), chronic treatment with fluoxetine completely abolished the hypothermic response and facilitated and strengthened the hyperthermic response. The effects of antidepressants on the responsiveness to LPS are probably not mediated by their effects on peripheral proinflammatory cytokine production, because LPS-induced expression of TNFalpha and IL-1beta mRNA in the spleen (assessed by semiquantitative in situ hybridization) was not altered following chronic treatment with either fluoxetine or imipramine. The effects of antidepressants on the acute phase response may have important clinical implications for the psychiatric and neuroendocrine disturbances that are commonly associated with various medical conditions.


Subject(s)
Antidepressive Agents/pharmacology , Behavior, Animal/drug effects , Brain/drug effects , Immune System/drug effects , Lipopolysaccharides/pharmacology , Neuroimmunomodulation/drug effects , Adrenal Glands/drug effects , Adrenal Glands/metabolism , Adrenergic Uptake Inhibitors/pharmacology , Animals , Behavior, Animal/physiology , Body Temperature/drug effects , Body Temperature/physiology , Brain/immunology , Brain/metabolism , Corticosterone/metabolism , Cytokines/genetics , Drug Administration Schedule/veterinary , Drug Interactions/physiology , Fluoxetine/pharmacology , Hypothalamo-Hypophyseal System/drug effects , Hypothalamo-Hypophyseal System/metabolism , Imipramine/pharmacology , Immune System/physiology , Male , Neuroimmunomodulation/physiology , RNA, Messenger/drug effects , RNA, Messenger/metabolism , Rats , Rats, Inbred F344 , Selective Serotonin Reuptake Inhibitors/pharmacology , Spleen/drug effects , Spleen/metabolism
19.
J Neurosci ; 21(3): 798-811, 2001 Feb 01.
Article in English | MEDLINE | ID: mdl-11157066

ABSTRACT

To test the hypothesis of an involvement of tachykinins in destabilization and hyperexcitation of neuronal circuits, gliosis, and neuroinflammation during cerebral ischemia, we investigated cell-specific expressional changes of the genes encoding substance P (SP), neurokinin B (NKB), and the tachykinin/neurokinin receptors (NK1, NK2, and NK3) after middle cerebral artery occlusion (MCAO) in the rat. Our analysis by quantitative in situ hybridization, immunohistochemistry, and confocal microscopy was concentrated on cerebrocortical areas that survive primary infarction but undergo secondary damage. Here, SP-encoding preprotachykinin-A and NK1 mRNA levels and SP-like immunoreactivity were transiently increased in GABAergic interneurons at 2 d after MCAO. Coincidently, MCAO caused a marked expression of SP and NK1 in a subpopulation of glutamatergic pyramidal cells, and in some neurons SP and NK1 mRNAs were coinduced. Elevated levels of the NKB-encoding preprotachykinin-B mRNA and of NKB-like immunoreactivity at 2 and 7 d after MCAO were confined to GABAergic interneurons. In parallel, the expression of NK3 was markedly downregulated in pyramidal neurons. MCAO caused transient NK1 expression in activated cerebrovenular endothelium within and adjacent to the infarct. NK1 expression was absent from activated astroglia or microglia. The differential ischemia-induced plasticity of the tachykinin system in distinct inhibitory and excitatory cerebrocortical circuits suggests that it may be involved in the balance of endogenous neuroprotection and neurotoxicity by enhancing GABAergic inhibitory circuits or by facilitating glutamate-mediated hyperexcitability. The transient induction of NK1 in cerebrovenular endothelium may contribute to ischemia-induced edema and leukocyte diapedesis. Brain tachykinin receptors are proposed as potential drug targets in stroke.


Subject(s)
Brain Ischemia/metabolism , Cerebral Cortex/metabolism , Endothelium, Vascular/metabolism , Receptors, Tachykinin/biosynthesis , Tachykinins/biosynthesis , Animals , Brain Ischemia/genetics , Brain Ischemia/pathology , Cerebral Cortex/blood supply , Cerebral Cortex/pathology , Cerebrovascular Circulation , Gene Expression Regulation , Glutamic Acid/metabolism , Infarction, Middle Cerebral Artery/genetics , Infarction, Middle Cerebral Artery/metabolism , Infarction, Middle Cerebral Artery/pathology , Male , Neurokinin B/genetics , Neurokinin B/metabolism , Neuronal Plasticity , Protein Precursors/genetics , Protein Precursors/metabolism , RNA, Messenger/metabolism , Rats , Rats, Long-Evans , Receptors, Neurokinin-1/genetics , Receptors, Neurokinin-1/metabolism , Receptors, Neurokinin-3/genetics , Receptors, Neurokinin-3/metabolism , Receptors, Tachykinin/genetics , Substance P/genetics , Substance P/metabolism , Tachykinins/genetics , Tachykinins/metabolism , Venules/metabolism , Venules/pathology , gamma-Aminobutyric Acid/metabolism
20.
J Histochem Cytochem ; 48(12): 1617-26, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11101630

ABSTRACT

Xenin is a 25-amino-acid peptide extractable from mammalian tissue. This peptide is biologically active. It stimulates exocrine pancreatic secretion and intestinal motility and inhibits gastric secretion of acid and food intake. Xenin circulates in the human plasma after meals. In this study, the cellular origin of xenin in the gastro-entero-pancreatic system of humans, Rhesus monkeys, and dogs was investigated by immunohistochemistry and immunoelectron microscopy. Sequence-specific antibodies against xenin detected specific endocrine cells in the duodenal and jejunal mucosa of all three species. These xenin-immunoreactive cells were distinct from enterochromaffin, somatostatin, motilin, cholecystokinin, neurotensin, and secretin cells, and comprised 8.8% of the chromogranin A-positive cells in the dog duodenum and 4.6% of the chromogranin A-positive cells in human duodenum. In all three species, co-localization of xenin was found with a subpopulation of gastric inhibitory polypeptide (GIP)-immunoreactive cells. Immunoelectron microscopy in the canine duodenal mucosa demonstrated accumulation of gold particles in round, homogeneous, and osmiophilic secretory granules with a closely adhering membrane of 187 +/- 19 nm diameter (mean +/- SEM). This cell type was found to be identical to the previously described canine GIP cell. Immunocytochemical expression of the peptide xenin in a subpopulation of chromogranin A-positive cells as well as the localization of xenin immunoreactivity in ultrastructurally characterized secretory granules permitted the identification of a novel endocrine cell type as the cellular source of circulating xenin.


Subject(s)
Duodenum , Intestinal Mucosa/metabolism , Peptides/metabolism , Amino Acid Sequence , Animals , Dogs , Gastric Inhibitory Polypeptide/metabolism , Gastrointestinal Hormones/immunology , Gastrointestinal Hormones/metabolism , Humans , Immune Sera , Immunohistochemistry , Intestinal Mucosa/cytology , Macaca mulatta , Microscopy, Electron , Microscopy, Immunoelectron , Molecular Sequence Data , Neurotensin , Peptides/immunology , Protein Precursors/immunology , Protein Precursors/metabolism , Species Specificity
SELECTION OF CITATIONS
SEARCH DETAIL
...