449 results on '"Belluardo,N."'
Search Results
102. [Growth and doubling time of the L1210 ascites tumor in DBA/2 mice after electrothermocoagulation of the tuberoinfundibular region of the hypothalamus]
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Bindoni M, Belluardo N, Ae, Marchese, Mudò G, Venera CARDILE, and Laguidara A
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Male ,Mice ,Mice, Inbred DBA ,Electrocoagulation ,Hypothalamus ,Animals ,Leukemia L1210 ,Cell Division
103. Hypothalamic control of the generation of mature natural killer lymphocytes in bone marrow and spleen of the mouse
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Belluardo, N., Mudo, G., Cardile, V., Graziella MIGLIORATI, Riccardi, C., Cella, S., and Bindoni, M.
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Male ,Hypothalamus ,Antibodies, Monoclonal ,Fluorescent Antibody Technique ,Hypothalamus, Middle ,Bone Marrow Cells ,Cytotoxicity Tests, Immunologic ,Hematopoietic Stem Cells ,Prolactin ,Killer Cells, Natural ,Mice, Inbred C57BL ,Mice ,Bone Marrow ,Growth Hormone ,Tumor Cells, Cultured ,Animals ,Interleukin-2 ,Corticosterone ,Spleen - Abstract
Following previous work showing that electrothermocoagulation of the median region of the hypothalamus (MH) caused a marked and permanent decrease in the cytotoxicity of natural killer (NK) cells and in the number of large granular lymphocytes, a study was made of the effect of such lesions on the generation of NK cells in the bone marrow (BM) and spleen of C57BL/6 mice. Fresh spleen and BM cells from MH-lesioned and sham-operated mice were cultured with 40 U of recombinant interleukin-2 (rIL-2). NK activity was significantly higher in BM of lesioned mice, whereas spleen NK activity was greater in the sham-operated controls. NK cells matured by culture with rIL-2 were characterized by assay with fluorescent monoclonal antibodies and found to display the typical NK phenotype. These results show that the number of NK precursors is greater in BM of MH-lesioned mice and that their migration into other organs is probably partially impeded. It can also be concluded that intactness of both BM and the hypothalamus is essential for the physiological generation of NK cells.
104. Nicotinic receptors and neurotrophic factors
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Belluardo, N., Mudo, G., Blum, M., and Fuxe, K.
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- 2000
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105. Regulation of FGF-2 gene by nicotinic receptors and its intracellular signaling mechanism in the rat brain
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Fuxe, K., Belluardo, N., Olsson, P.-A., Mudo, G., Liu, F.-L., and Changeux, J.-P.
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- 2002
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106. Changes in gene expression of AMPA-selective glutamate receptor subunits induced by status epilepticus in rat brain
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Condorelli, D. F., Belluardo, N., Mudo, G., and Dell'Albani, P.
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- 1994
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107. Crossfostering and early development of natural killer cytotoxic activity in various inbred mouse strains
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Belluardo, N., Mudo, G., Campisi, A., and Vanella, A.
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- 1993
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108. Neurotoxic injury in rat hippocampus differentially affects multiple trkB and trkC transcripts
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Belluardo, N., Salin, T., Dell'Albani, P., and Mudo, G.
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- 1995
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109. Developmental regulation of brain-derived neurotrophic factor messenger RNAs transcribed from different promoters in the rat brain
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Timmusk, T., Belluardo, N., Persson, H., and Metsis, M.
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- 1994
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110. Existence of muscarinic acetylcholine receptor (mAChR) and fibroblast growth factor receptor (FGFR) heteroreceptor complexes and their enhancement of neurite outgrowth in neural hippocampal cultures.
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Di Liberto, V., Borroto-Escuela, D.O., Frinchi, M., Verdi, V., Fuxe, K., Belluardo, N., and Mudò, G.
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CHOLINERGIC receptors , *MUSCARINIC agonists , *ACETYLCHOLINE , *ACETYLCHOLINE-binding proteins , *NOGO protein - Abstract
Background Recently, it was demonstrated that G-protein-coupled receptors (GPCRs) can transactivate tyrosine kinase receptors in absence of their ligands. In this work, driven by the observation that mAChRs and fibroblast growth factor receptors (FGFRs) share signalling pathways and regulation of brain functions, it was decided to explore whether mAChRs activation may transactivate FGFRs and, if so, to characterize the related trophic effects in cultured hippocampal neurons. Methods Oxotremorine-M transactivation of FGFRs and related trophic effects were tested in primary hippocampal neurons. Western blotting and in situ proximity ligation assay (PLA) were used to detect FGFR phosphorylation (pFGFR) levels and M 1 R-FGFR1 heteroreceptor complexes, respectively. Results Oxotremorine-M, a non-selective mAChRs agonist, was able to transactivate FGFR and this transactivation was blocked by Src inhibitors. Oxotremorine-M treatment produced a significant increase in the primary neurite outgrowth that was blocked by pre-treatment with the pFGFR inhibitor SU5402 and Src inhibitors. This trophic effect was almost similar to that induced by fibroblast growth factor-2 (FGF-2). By using atropine as nonselective mAChRs or pirenzepine as selective antagonist for M 1 receptor (M 1 R) we could show that mAChRs are involved in modulating the pFGFRs. Using PLA, M 1 R-FGFR1 heteroreceptor complexes were identified in the hippocampus and cerebral cortex. Conclusion The current findings, by showing functional mAChR-FGFR interactions, will contribute to advance the understanding of the mechanisms involved in the actions of cholinergic drugs on neuronal plasticity. General significant Data may help to develop novel therapeutic strategies not only for neurodegenerative diseases but also for depression-induced atrophy of hippocampal neurons. [ABSTRACT FROM AUTHOR]
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- 2017
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111. Persistence on Anti-Tumour Necrosis Factor Therapy in Older Patients with Inflammatory Bowel Disease Compared with Younger Patients: Data from the Sicilian Network for Inflammatory Bowel Diseases (SN-IBD)
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Fabio Salvatore Macaluso, Gaetano Inserra, Marco Ventimiglia, Maria Cappello, Mario Cottone, Antonio Carroccio, Angela Alibrandi, Antonino Carlo Privitera, Walter Fries, Filippo Mocciaro, N. Belluardo, A. Magnano, Sara Renna, S. Siringo, S. Garufi, Serena Porcari, C. Ferracane, Ambrogio Orlando, Alessandro Vitello, C. Bertolami, Oriana Fidanza, Roberto Di Mitri, Anna Viola, G. Magrì, and Porcari S, Viola A, Orlando A, Privitera AC, Ferracane C, Cappello M, Vitello A, Siringo S, Inserra G, Magnano A, Mocciaro F, Di Mitri R, Belluardo N, Fidanza O, Garufi S, Magrì G, Bertolami C, Carroccio A, Macaluso FS, Renna S, Ventimiglia M, Alibrandi A, Cottone M, Fries W
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Adult ,Male ,medicine.medical_specialty ,Settore MED/09 - Medicina Interna ,Drug-Related Side Effects and Adverse Reactions ,Kaplan-Meier Estimate ,Anti-Tumour Necrosis Factor ,Disease ,Inflammatory bowel disease ,Cohort Studies ,03 medical and health sciences ,0302 clinical medicine ,Pharmacotherapy ,Internal medicine ,medicine ,Humans ,Pharmacology (medical) ,Treatment Failure ,030212 general & internal medicine ,Adverse effect ,Aged ,Retrospective Studies ,Aged, 80 and over ,Tumor Necrosis Factor-alpha ,business.industry ,Inflammatory Bowel Disease ,Adalimumab ,Age Factors ,Antibodies, Monoclonal ,Retrospective cohort study ,Middle Aged ,Inflammatory Bowel Diseases ,medicine.disease ,Ulcerative colitis ,Infliximab ,Sicilian Network for Inflammatory Bowel Diseases (SN-IBD) ,Withholding Treatment ,Concomitant ,Female ,Geriatrics and Gerontology ,business ,030217 neurology & neurosurgery ,Cohort study - Abstract
BACKGROUND AND OBJECTIVE: Older people with inflammatory bowel disease (IBD) appear to have a lower response to anti-tumour necrosis factor (TNF) therapy, with more frequent complications than younger patients. The objective of this study was to assess persistence on therapy and the safety of anti-TNF therapy in older patients (aged ≥ 60 years). METHODS: We retrospectively reviewed the database of the Sicilian Network for Inflammatory Bowel Diseases (SN-IBD), extracting data regarding IBD patients aged ≥ 60 years and controls
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- 2020
112. Investigating the Role of Guanosine on Human Neuroblastoma Cell Differentiation and the Underlying Molecular Mechanisms
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Natale Belluardo, Giuseppa Mudò, Valentina Di Liberto, Monica Frinchi, Daniele F. Condorelli, Ugo Traversa, Francisco Ciruela, Renata Ciccarelli, Patrizia Di Iorio, Patricia Giuliani, Belluardo N., Mudo' G., Di Liberto V., Frinchi M., Condorelli D.F., Traversa U., Ciruela F., Ciccarelli R., Di Iorio P., and Giuliani P.
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Neurite ,Cellular differentiation ,Guanosine ,Purine nucleoside phosphorylase ,RM1-950 ,Nucleoside transporter ,Settore BIO/09 - Fisiologia ,chemistry.chemical_compound ,neuroblastoma ,guanine, guanosine, guanylate cyclase, heme oxygenase, neuroblastoma, protein kinase C, purine nucleoside phosphorylase, SH-SY5Ydifferentiation ,Nucleòsids ,Extracellular ,Pharmacology (medical) ,guanine ,Pharmacology ,biology ,Marcadors tumorals ,Nucleosides ,SH-SY5Ydifferentiation ,Brief Research Report ,heme oxygenase ,purine nucleoside phosphorylase ,Cell biology ,guanylate cyclase ,guanosine ,chemistry ,Cell culture ,Tumor markers ,Settore BIO/14 - Farmacologia ,biology.protein ,Therapeutics. Pharmacology ,Nucleoside ,protein kinase C - Abstract
Neuroblastoma arises from neural crest cell precursors failing to complete the process of differentiation. Thus, agents helping tumor cells to differentiate into normal cells can represent a valid therapeutic strategy. Here, we evaluated whether guanosine (GUO), a natural purine nucleoside, which is able to induce differentiation of many cell types, may cause the differentiation of human neuroblastoma SH-SY5Y cells and the molecular mechanisms involved. We found that GUO, added to the cell culture medium, promoted neuron-like cell differentiation in a time- and concentration-dependent manner. This effect was mainly due to an extracellular GUO action since nucleoside transporter inhibitors reduced but not abolished it. Importantly, GUO-mediated neuron-like cell differentiation was independent of adenosine receptor activation as it was not altered by the blockade of these receptors. Noteworthy, the neuritogenic activity of GUO was not affected by blocking the phosphoinositide 3-kinase pathway, while it was reduced by inhibitors of protein kinase C or soluble guanylate cyclase. Furthermore, the inhibitor of the enzyme heme oxygenase-1 but not that of nitric oxide synthase reduced GUO-induced neurite outgrowth. Interestingly, we found that GUO was largely metabolized into guanine by the purine nucleoside phosphorylase (PNP) enzyme released from cells. Taken together, our results suggest that GUO, promoting neuroblastoma cell differentiation, may represent a potential therapeutic agent; however, due to its spontaneous extracellular metabolism, the role played by the GUO-PNP-guanine system needs to be further investigated.
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- 2021
113. Guanosine-Mediated Anxiolytic-Like Effect: Interplay with Adenosine A1 and A2A Receptors
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Renata Ciccarelli, Giuseppa Mudò, Patrizia Di Iorio, Roberta Garozzo, Fulvio Plescia, Daniele F. Condorelli, Monica Frinchi, Francisco Ciruela, Natale Belluardo, Valentina Di Liberto, Francesco Caciagli, Vincenzo Verdi, Maria Grillo, Università degli studi di Palermo - University of Palermo, Institut de psychiatrie et neurosciences de Paris (IPNP - U1266 Inserm), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Universitat de Barcelona (UB), University of Catania [Italy], University 'G. d'Annunzio' of Chieti-Pescara [Chieti and Pescara, Italy], Frinchi M., Verdi V., Plescia F., Ciruela F., Grillo M., Garozzo R., Condorelli D.F., Di Iorio P., Caciagli F., Ciccarelli R., Belluardo N., Di Liberto V., Mudo' G., Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), and Martinez Rico, Clara
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Light ,Pharmacology ,Anxiety ,Settore BIO/09 - Fisiologia ,Hippocampus ,lcsh:Chemistry ,chemistry.chemical_compound ,0302 clinical medicine ,Receptor ,lcsh:QH301-705.5 ,Spectroscopy ,caffeine ,0303 health sciences ,Behavior, Animal ,General Medicine ,Darkness ,3. Good health ,Computer Science Applications ,adenosine ,CCPA ,[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,A1R ,Caffeine ,medicine.drug ,Receptor, Adenosine A2A ,Guanosine ,Catalysis ,Article ,Inorganic Chemistry ,03 medical and health sciences ,medicine ,Animals ,[SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,Physical and Theoretical Chemistry ,Binding site ,2A ,Molecular Biology ,030304 developmental biology ,Dose-Response Relationship, Drug ,Receptor, Adenosine A1 ,behavior ,Organic Chemistry ,Cell Membrane ,Antagonist ,Adenosine ,Adenosine receptor ,Rats ,guanosine ,A2AR ,lcsh:Biology (General) ,lcsh:QD1-999 ,chemistry ,030217 neurology & neurosurgery - Abstract
Acute or chronic administration of guanosine (GUO) induces anxiolytic-like effects, for which the adenosine (ADO) system involvement has been postulated yet without a direct experimental evidence. Thus, we aimed to investigate whether adenosine receptors (ARs) are involved in the GUO-mediated anxiolytic-like effect, evaluated by three anxiety-related paradigms in rats. First, we confirmed that acute treatment with GUO exerts an anxiolytic-like effect. Subsequently, we investigated the effects of pretreatment with ADO or A1R (CPA, CCPA) or A2AR (CGS21680) agonists 10 min prior to GUO on a GUO-induced anxiolytic-like effect. All the combined treatments blocked the GUO anxiolytic-like effect, whereas when administered alone, each compound was ineffective as compared to the control group. Interestingly, the pretreatment with nonselective antagonist caffeine or selective A1R (DPCPX) or A2AR (ZM241385) antagonists did not modify the GUO-induced anxiolytic-like effect. Finally, binding assay performed in hippocampal membranes showed that [3H]GUO binding became saturable at 100&ndash, 300 nM, suggesting the existence of a putative GUO binding site. In competition experiments, ADO showed a potency order similar to GUO in displacing [3H]GUO binding, whereas AR selective agonists, CPA and CGS21680, partially displaced [3H]GUO binding, but the sum of the two effects was able to displace [3H]GUO binding to the same extent of ADO alone. Overall, our results strengthen previous data supporting GUO-mediated anxiolytic-like effects, add new evidence that these effects are blocked by A1R and A2AR agonists and pave, although they do not elucidate the mechanism of GUO and ADO receptor interaction, for a better characterization of GUO binding sites in ARs.
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- 2020
114. Group II metabotropic glutamate receptor activation by agonist LY379268 treatment increases the expression of brain derived neurotrophic factor in the mouse brain
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Di Liberto, V., Bonomo, A., Frinchi, M., Belluardo, N., and Mudò, G.
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GLUTAMIC acid , *NEURAL receptors , *NEUROTROPHINS , *LABORATORY rats , *NEUROPROTECTIVE agents , *CELLULAR signal transduction , *IN situ hybridization , *NERVE growth factor - Abstract
Abstract: A number of in vitro and in vivo studies using selective agonists have indicated a neuroprotective role for group-II metabotropic glutamate (mGlu2/3) receptors in various models of neuronal injury. Although an interplay among neurotrophic factors and mGlu2/3 receptors signalling system has been suggested as possible mechanism involved on neuroprotection, at present poor information are available concerning the in vivo regulation by mGlu2/3 receptors activation of specific neurotrophic factors. By using in situ hybridization and western blotting methods the aim of present study was to analyse the potential regulatory role of selective mGluR2/3 agonist LY379268 treatment on brain derived neurotrophic factor (BDNF) expression in the mouse brain. The treatment with LY379268 evidenced a significant upregulation of BDNF mRNA levels in the cerebral cortex and in the hippocampal formation with a peak at 3 h from treatment and its disappearance already at 6 h from treatment. An analysis of dose-effect curve revealed that LY379268 may significantly enhance BDNF mRNA expression already at dose of 0.250 mg/kg b.w. The upregulation of BDNF mRNA expression was followed by a significant increase of BDNF protein levels at 24 h from LY379268 treatment. These effects of LY379268 treatment on BDNF expression were restricted to neuronal cells and were blocked by the new selective mGlu2/3 receptor antagonist LY341495, suggesting a receptor specificity. Taken together these findings suggest that several previous observed neuroprotective and trophic actions of mGluR2/3 agonists treatment may be mediated, at least in the cerebral cortex and hippocampal formation, by upregulation of BDNF expression. [Copyright &y& Elsevier]
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- 2010
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115. Existence of muscarinic acetylcholine receptor (mAChR) and fibroblast growth factor receptor (FGFR) heteroreceptor complexes and their enhancement of neurite outgrowth in neural hippocampal cultures
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Kjell Fuxe, Monica Frinchi, Vincenzo Verdi, Giuseppa Mudò, V. Di Liberto, Natale Belluardo, D.O. Borroto-Escuela, DI LIBERTO, V., Borroto-Escuela, D., Frinchi, M., Verdi, V., Fuxe, K., Belluardo, N., and Mudo', G.
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Male ,0301 basic medicine ,Hippocampus ,Biochemistry ,Receptor tyrosine kinase ,Receptors, G-Protein-Coupled ,Rats, Sprague-Dawley ,Transactivation ,0302 clinical medicine ,Muscarinic acetylcholine receptor ,Neural plasticity ,Neurons ,Neuronal Plasticity ,biology ,Receptors, Muscarinic ,Cell biology ,Fibroblast growth factor receptor ,Fibroblast Growth Factor 2 ,Signal Transduction ,Proto-oncogene tyrosine-protein kinase Src ,medicine.medical_specialty ,Neurite ,Neuronal Outgrowth ,Biophysics ,Heteroreceptor ,03 medical and health sciences ,Hippocampu ,Internal medicine ,medicine ,Animals ,Receptor, Fibroblast Growth Factor, Type 1 ,Rats, Wistar ,Molecular Biology ,Animal ,Oxotremorine ,Fibroblast growth factor receptor 1 ,Receptor, Muscarinic M1 ,Neuron ,Receptors, Fibroblast Growth Factor ,Rats ,FGFR1 ,030104 developmental biology ,Endocrinology ,M1receptor ,Biophysic ,Heteroreceptor complexe ,biology.protein ,Rat ,030217 neurology & neurosurgery - Abstract
Background Recently, it was demonstrated that G-protein-coupled receptors (GPCRs) can transactivate tyrosine kinase receptors in absence of their ligands. In this work, driven by the observation that mAChRs and fibroblast growth factor receptors (FGFRs) share signalling pathways and regulation of brain functions, it was decided to explore whether mAChRs activation may transactivate FGFRs and, if so, to characterize the related trophic effects in cultured hippocampal neurons. Methods Oxotremorine-M transactivation of FGFRs and related trophic effects were tested in primary hippocampal neurons. Western blotting and in situ proximity ligation assay (PLA) were used to detect FGFR phosphorylation (pFGFR) levels and M1R-FGFR1 heteroreceptor complexes, respectively. Results Oxotremorine-M, a non-selective mAChRs agonist, was able to transactivate FGFR and this transactivation was blocked by Src inhibitors. Oxotremorine-M treatment produced a significant increase in the primary neurite outgrowth that was blocked by pre-treatment with the pFGFR inhibitor SU5402 and Src inhibitors. This trophic effect was almost similar to that induced by fibroblast growth factor-2 (FGF-2). By using atropine as nonselective mAChRs or pirenzepine as selective antagonist for M1 receptor (M1R) we could show that mAChRs are involved in modulating the pFGFRs. Using PLA, M1R-FGFR1 heteroreceptor complexes were identified in the hippocampus and cerebral cortex. Conclusion The current findings, by showing functional mAChR-FGFR interactions, will contribute to advance the understanding of the mechanisms involved in the actions of cholinergic drugs on neuronal plasticity. General significant Data may help to develop novel therapeutic strategies not only for neurodegenerative diseases but also for depression-induced atrophy of hippocampal neurons.
- Published
- 2017
116. Connexin-30 mRNA Is Up-Regulated in Astrocytes and Expressed in Apoptotic Neuronal Cells of Rat Brain Following Kainate-Induced Seizures
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Condorelli, D. F., Mudò, G., Trovato-Salinaro, A., Mirone, M. B., Amato, G., and Belluardo, N.
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ASTROCYTES , *IMMUNOHISTOCHEMISTRY - Abstract
Glial connexins (Cxs) make an extensively interconnected functional syncytium created by a network of gap junctions between astrocytes and oligodendrocytes. Among Cxs expressed in the brain, Cx30 is expressed in grey matter astrocytes, as shown at the protein level by immunoistochemistry. In the present study we aimed to perform a detailed study of the regional distribution of Cx30 mRNA in the adult and postnatal developing rat brain, analyzing its expression by in situ hybridization, and determining its cell type localization by double labeling. Recently, it has been suggested that neuronal activity may control the level of intercellular communication between astrocytes through gap junctions channels. Thus, a second aim of the present study was to investigate the short-term effects of kainate-induced seizures on Cx30 expression. The results showed that, in basal condition, Cx30 was expressed only in grey matter astrocytes with distinct regional patterns in developing and adult brain. Kainate treatment induced strong and region-specific changes of astroglial Cx30 mRNA levels and expression of Cx30 mRNA in neuronal cells undergoing cell death, suggesting a direct or indirect involvement of this connexin in the neuronal apoptotic process. [Copyright &y& Elsevier]
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- 2002
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117. Anti-inflammatory and antioxidant effects of muscarinic acetylcholine receptor (mAChR) activation in the rat hippocampus
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Monica Frinchi, Marta Di Carlo, Pietro Scaduto, Maria Fatima Massenti, Natale Belluardo, Domenico Nuzzo, Giuseppa Mudò, Frinchi M., Nuzzo D., Scaduto P., Di Carlo M., Massenti M.F., Belluardo N., and Mudo G.
- Subjects
Male ,Hydrocortisone ,medicine.medical_treatment ,Interleukin-1beta ,Neuroimmunology ,Anti-Inflammatory Agents ,lcsh:Medicine ,Pharmacology ,medicine.disease_cause ,Hippocampus ,Settore BIO/09 - Fisiologia ,Antioxidants ,Superoxide Dismutase-1 ,Muscarinic acetylcholine receptor ,Phosphorylation ,lcsh:Science ,chemistry.chemical_classification ,Multidisciplinary ,biology ,neurodegeneration ,Alzheimer's disease ,Receptors, Muscarinic ,Neuroprotective Agents ,Cytokine ,Signal Transduction ,medicine.drug ,Restraint, Physical ,Agonist ,medicine.drug_class ,Scopolamine ,muscarinic acetylcholine receptor ,Muscarinic Agonists ,Article ,Oxotremorine, anti-inflammatory cytokines ,Superoxide dismutase ,Heat shock protein ,Oxotremorine ,medicine ,Animals ,Rats, Wistar ,Inflammation ,Reactive oxygen species ,Interleukin-6 ,Superoxide Dismutase ,lcsh:R ,Transcription Factor RelA ,Rats ,Oxidative Stress ,chemistry ,biology.protein ,lcsh:Q ,Reactive Oxygen Species ,Protein Processing, Post-Translational ,Oxidative stress - Abstract
Recently we found that acute treatment with Oxotremorine (Oxo), a non-selective mAChRs agonist, up-regulates heat shock proteins and activates their transcription factor heat shock factor 1 in the rat hippocampus. Here we aimed to investigate: a) if acute treatment with Oxo may regulate pro-inflammatory or anti-inflammatory cytokines and oxidative stress in the rat hippocampus; b) if chronic restraint stress (CRS) induces inflammatory or oxidative alterations in the hippocampus and whether such alterations may be affected by chronic treatment with Oxo. In the acute experiment, rats were injected with single dose of Oxo (0.4 mg/kg) and sacrificed at 24 h, 48 h and 72 h. In the CRS experiment, the rats were exposed for 21 days to the CRS and then were treated with Oxo (0.2 mg/kg) for further 10 days. The acute Oxo treatment showed an ability to significantly reduce reactive oxygen species (ROS), singlet oxygen (1O2), pro-inflammatory cytokines levels (IL-1β and IL-6) and phosphorylated NF-κB-p65. Acute Oxo treatment also increased superoxide dismutase (SOD)-2 protein levels and stimulated SOD activity. No differences were detected in the anti-inflammatory cytokine levels, including IL-10 and TGF-β1. In the group of rats exposed to the CRS were found increased hippocampal IL-1β and IL-6 levels, together with a reduction of SOD activity level. These changes produced by CRS were counteracted by chronic Oxo treatment. In contrast, the upregulation of ROS and 1O2 levels in the CRS group was not counteracted by chronic Oxo treatment. The results revealed a hippocampal anti-inflammatory and antioxidant effect of Oxo treatment in both basal conditions and anti-inflammatory in the CRS rat model.
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- 2019
118. A Propensity Score-matched Comparison of Infliximab and Adalimumab in Tumour Necrosis Factor-α Inhibitor-naïve and Non-naïve Patients with Crohn's Disease: Real-Life Data from the Sicilian Network for Inflammatory Bowel Disease
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A.C. Privitera, Salvatore Genova, Michele Citrano, G. Magrì, S. Siringo, Maria Cappello, Antonio Carroccio, Mario Cottone, Fabio Salvatore Macaluso, Filippo Mocciaro, Giulia Rizzuto, A. Magnano, G. Scarpulla, Sara Renna, N. Belluardo, Ambrogio Orlando, Gaetano Inserra, Claudio Romano, C. Ferracane, Salvatore Accomando, C. Bertolami, Walter Fries, R. Vassallo, R. Orlando, Roberto Di Mitri, A. Trovatello, Serena Porcari, Marco Ventimiglia, MacAluso F.S., Fries W., Privitera A.C., Cappello M., Siringo S., Inserra G., Magnano A., Di Mitri R., Mocciaro F., Belluardo N., Scarpulla G., Magri G., Trovatello A., Carroccio A., Genova S., Bertolami C., Vassallo R., Romano C., Citrano M., Accomando S., Ventimiglia M., Renna S., Orlando R., Rizzuto G., Porcari S., Ferracane C., Cottone M., and Orlando A.
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0301 basic medicine ,Adult ,Male ,medicine.medical_specialty ,Inflammatory bowel disease ,Gastroenterology ,03 medical and health sciences ,Young Adult ,0302 clinical medicine ,Crohn Disease ,Gastrointestinal Agents ,Internal medicine ,medicine ,Adalimumab ,Humans ,Young adult ,Sicily ,propensity score ,Crohn's disease ,business.industry ,Tumor Necrosis Factor-alpha ,General Medicine ,Odds ratio ,medicine.disease ,infliximab ,Female ,Infliximab ,Propensity Score ,Treatment Outcome ,030104 developmental biology ,Cohort ,Propensity score matching ,030211 gastroenterology & hepatology ,business ,medicine.drug - Abstract
Background and aims There is an unmet need to better understand the effectiveness of different biologics in inflammatory bowel diseases. We aimed at performing a multicentre, real-life comparison of the effectiveness of infliximab [IFX] and adalimumab [ADA] in Crohn's disease [CD]. Methods Data of consecutive patients with CD treated with IFX and ADA from January 2013 to May 2017 were extracted from the cohort of the Sicilian Network for Inflammatory Bowel Disease. We used propensity score-matching accounting for the main baseline characteristics in TNF-α inhibitor-naive and non-naive patients. Results A total of 632 patients [735 total treatments] were included. Among naive patients, a clinical benefit [the sum of steroid-free remission plus clinical response] was achieved in 81.8% patients treated with ADA and in 77.6% patients treated with IFX (adjusted odds ratio [OR]: 1.23, 95% CI 0.63-2-44, p = 0.547] at 12 weeks; after 1 year, a clinical benefit was achieved in 69.2% of patients treated with ADA and in 64.5% patients treated with IFX [adjusted OR: 1.10, 95% CI 0.61-1.96, p = 0.766]. Among non-naive patients, a clinical benefit was achieved in 61.7% of patients treated with ADA and in 68.1% of patients treated with IFX [adjusted OR: 0.72, 95% CI 0.21-2.44, p = 0.600] at 12 weeks; after 1 year, a clinical benefit was achieved in 48.9% of patients treated with ADA and in 40.4% patients treated with IFX [adjusted OR: 1.23, 95% CI 0.54-2.86, p = 0.620]. Conclusions In this propensity score-matched comparison of ADA and IFX in CD, both drugs showed high rates of clinical benefit, without significant differences between them.
- Published
- 2019
119. Nicotine and nicotinic receptors in neuronal injury. Focus on basic FGF mechanisms
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Belluardo, N., Blum, M., Mudo, G., and Fuxe, K.
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- 1998
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120. Detection, Analysis, and Quantification of GPCR Homo- and Heteroreceptor Complexes in Specific Neuronal Cell Populations Using the In Situ Proximity Ligation Assay
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Miguel Pérez de la Mora, Sarah Beggiato, Kjell Fuxe, Gemma Navarro, Thorsten Schaefer, Małgorzata Filip, Michael Di Palma, Giuseppa Mudò, Irene Reyes-Resina, Sergio Tanganelli, Kirill Shumilov, Rafael Franco, Patrizia Ambrogini, Harriët Schellekens, Kristina Friedland, Luca Ferraro, Karolina Wydra, Ismael Valladolid-Acebes, Dasiel O. Borroto-Escuela, Natale Belluardo, Stefano Sartini, Manuel Narváez, Kjell Fuxe, Dasiel O. Borroto-Escuela, Borroto-Escuela D.O., Narvaez M., Valladolid-Acebes I., Shumilov K., Di Palma M., Wydra K., Schaefer T., Reyes-Resina I., Navarro G., Mudo' G., Filip M., Sartini S., Friedland K., Schellekens H., Beggiato S., Ferraro L., Tanganelli S., Franco R., Belluardo N., Ambrogini P., Perez de la Mora M., and Fuxe K.
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0301 basic medicine ,In situ ,In situ proximity ligation assay ,Chemistry ,Cell ,Proximity ligation assay ,Heteroreceptor ,Settore BIO/09 - Fisiologia ,Immunohistochemistry ,Receptor–receptor interaction ,Stoichiometry ,NO ,G protein-coupled receptors, Immunohistochemistry, In situ proximity ligation assay, Heteroreceptor complexes, Dimerization, Receptor–receptor interaction, Stoichiometry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,G protein-coupled receptors ,Biophysics ,medicine ,Heteroreceptor complexes ,Dimerization ,030217 neurology & neurosurgery ,G protein-coupled receptor - Abstract
GPCR’s receptosome operates via coordinated changes between the receptor expression, their modifications and interactions between each other. Perturbation in specific heteroreceptor complexes and/or their balance/equilibrium with other heteroreceptor complexes and corresponding homoreceptor complexes is considered to have a role in pathogenic mechanisms. Such mechanisms lead to mental and neurological diseases, including drug addiction, depression, Parkinson’s disease, and schizophrenia. To understand the associations of GPCRs and to unravel the global picture of their receptor–receptor interactions in the brain, different experimental detection techniques for receptor–receptor interactions have been established (e.g., co-immunoprecipitation based approach). However, they have been criticized for not reflecting the cellular situation or the dynamic nature of receptor–receptor interactions. Therefore, the detection and visualization of GPCR homo- and heteroreceptor complexes in the brain remained largely unknown until recent years, when a well-characterized in situ proximity ligation assay (in situ PLA) was adapted to validate the receptor complexes in their native environment. The in situ PLA protocol presented here can be used to visualize GPCR receptor–receptor interactions in cells and tissues in a highly sensitive and specific manner. We have developed a combined method using immunohistochemistry and PLA, particularly aimed to monitor interactions between GPCRs in specific neuronal cell populations. This allows the analysis of homo- and heteroreceptor complexes at a cellular and subcellular level. The method has the advantage that it can be used in clinical specimens, providing localized, quantifiable homo- and heteroreceptor complexes detected in single cells. We compare the advantages and limitations of the methods, underlining recent progress and the growing importance of these techniques in basic research. We discuss also their potential as tools for drug development and diagnostics.
- Published
- 2018
121. Detection of Fibroblast Growth Factor Receptor 1 (FGFR1) Transactivation by Muscarinic Acetylcholine Receptors (mAChRs) in Primary Neuronal Hippocampal Cultures Through Use of Biochemical and Morphological Approaches
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Kjell Fuxe, Natale Belluardo, Dasiel O. Borroto-Escuela, Valentina Di Liberto, Giuseppa Mudò, Fuxe, K., Borroto-Escuela D.O., Di Liberto V., Mudo' G., Borroto-Escuela D.O., Fuxe K., and Belluardo N.
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Transactivation ,Chemistry ,Fibroblast growth factor receptor 1 ,Tyrosine kinase receptor ,Hippocampal formation ,Hippocampus ,Settore BIO/09 - Fisiologia ,Fibroblast growth factor receptor ,Western blotting ,Cell biology ,Muscarinic acetylcholine receptor ,Primary neuronal culture ,Neurite growth ,Phosphorylation ,Receptor–receptor interactions - Abstract
In addition to their canonical intracellular signals involved in the regulation of neuronal plasticity, G-protein coupled receptors can also rapidly transactivate tyrosine kinase receptors and their downstream intracellular signaling in absence of specific ligands. Here we describe our protocol for dissociating and maintaining hippocampal primary neurons in high- and low-density culture, followed by a description of methods employed to evaluate neurite outgrowth and protein phosphorylation associated with fibroblast growth factor receptor 1 transactivation by muscarinic acetylcholine receptors. Our goal was to provide the reader with detailed protocols of the abovementioned techniques and to highlight advantages and limitations of the used approaches as compared to other valid alternatives.
- Published
- 2018
122. Altered gastrointestinal motility in an animal model of Lesch-Nyhan disease
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Patrizia Di Iorio, Hyder A. Jinnah, Daniele F. Condorelli, Flavia Mulè, Renata Ciccarelli, Domenico Nuzzo, Giuseppa Mudò, Rosa Serio, Francesco Caciagli, Natale Belluardo, Maria Grazia Zizzo, Monica Frinchi, Zizzo, M., Frinchi, M., Nuzzo, D., Jinnah, H., Mudò, G., Condorelli, D., Caciagli, F., Ciccarelli, R., Di Iorio, P., Mulè, F., Belluardo, N., and Serio, R.
- Subjects
0301 basic medicine ,Atropine ,Male ,Hypoxanthine Phosphoribosyltransferase ,Lesch-Nyhan Syndrome ,Dopamine ,medicine.disease_cause ,Settore BIO/09 - Fisiologia ,Lesch-Nyhan ,Mice ,0302 clinical medicine ,Enzyme Inhibitors ,Evoked Potentials ,Myenteric plexus ,HGprt deficient mice ,Neurotransmitter Agents ,Brain ,NG-Nitroarginine Methyl Ester ,Knockout mouse ,Cytokines ,Acetylcholine ,medicine.drug ,medicine.medical_specialty ,Carbachol ,Tyrosine 3-Monooxygenase ,Colon ,Motility ,Mice, Transgenic ,In Vitro Techniques ,Endocrine and Autonomic System ,Article ,Contractility ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Internal medicine ,medicine ,Animals ,Cytokine ,Endocrine and Autonomic Systems ,business.industry ,Muscle, Smooth ,Benzazepines ,Mice, Inbred C57BL ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Gene Expression Regulation ,HGprt enzyme ,Face ,Oxidative stre ,Neurology (clinical) ,Lipid Peroxidation ,business ,Gastrointestinal Motility ,Reactive Oxygen Species ,030217 neurology & neurosurgery ,Oxidative stress - Abstract
Mutations in the HGPRT1 gene, which encodes hypoxanthine-guanine phosphoribosyltransferase (HGprt), housekeeping enzyme responsible for recycling purines, lead to Lesch-Nyhan disease (LND). Clinical expression of LND indicates that HGprt deficiency has adverse effects on gastrointestinal motility. Therefore, we aimed to evaluate intestinal motility in HGprt knockout mice (HGprt(−)). Spontaneous and neurally evoked mechanical activity was recorded in vitro as changes in isometric tension in circular muscle strips of distal colon. HGprt(−) tissues showed a lower in amplitude spontaneous activity and atropine-sensitivity neural contraction compared to control mice. The responses to carbachol and to high KCl were reduced, demonstrating a widespread impairment of contractility. L-NAME was not able in the HGprt(−) tissues to restore the large amplitude contractile activity typical of control. In HGprt(−) colon, a reduced expression of dopaminergic D1 receptor was observed together with the loss of its tonic inhibitory activity present in control-mice. The analysis of inflammatory and oxidative stress in colonic tissue of HGprt(−) mice revealed a significant increase of lipid peroxidation associated with over production of oxygen free radicals. In conclusion, HGprt deficiency in mice is associated with a decrease in colon contractility, not dependent upon reduction of acetylcholine release from the myenteric plexus or hyperactivity of inhibitory signalling. By contrast the increased levels of oxidative stress could partially explain the reduced colon motility in HGprt(−) mice. Colonic dysmotility observed in HGprt(−) mice may mimic the gastrointestinal dysfunctions symptoms of human syndrome, providing a useful animal model to elucidate the pathophysiology of this problem in the LND.
- Published
- 2017
123. Mild Aerobic Exercise Training Hardly Affects the Diaphragm of mdx Mice
- Author
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Giuseppe, Morici, Monica, Frinchi, Alessandro, Pitruzzella, Valentina, Di Liberto, Rosario, Barone, Andrea, Pace, Valentina, Di Felice, Natale, Belluardo, Francesco, Cappello, Giuseppa, Mudò, Maria R, Bonsignore, Morici, G., Frinchi, M., Pitruzzella, A., Di Liberto, V., Barone, R., Pace, A., Di Felice, V., Belluardo, N., Cappello, F., Mudò, G., and Bonsignore, M.
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Male ,Duchenne muscular dystrophy ,chaperonin ,Time Factors ,Diaphragm ,Settore MED/10 - Malattie Dell'Apparato Respiratorio ,Settore BIO/09 - Fisiologia ,Connexins ,Mitochondrial Proteins ,Necrosis ,endurance training ,Animals ,Genetic Predisposition to Disease ,HSP70 Heat-Shock Proteins ,stress markers ,Muscle Strength ,NF-kB ,Settore BIO/16 - Anatomia Umana ,Transcription Factor RelA ,Chaperonin 60 ,Settore CHIM/06 - Chimica Organica ,Exercise Therapy ,Muscular Dystrophy, Duchenne ,Disease Models, Animal ,Phenotype ,Mice, Inbred mdx ,Physical Endurance - Abstract
In the mdx mice model of Duchenne Muscular Dystrophy (DMD), mild endurance exercise training positively affected limb skeletal muscles, whereas few and controversial data exist on the effects of training on the diaphragm. The diaphragm was examined in mdx (C57BL/10ScSn-Dmdmdx) and wild-type (WT, C57BL/10ScSc) mice under sedentary conditions (mdx-SD, WT-SD) and during mild exercise training (mdx-EX, WT-EX). At baseline, and after 30 and 45 days (training: 5 d/wk for 6 weeks), diaphragm muscle morphology and Cx39 protein were assessed. In addition, tissue levels of the chaperonins Hsp60 and Hsp70 and the p65 subunit of nuclear factor-kB (NF-kB) were measured in diaphragm, gastrocnemius, and quadriceps in each experimental group at all time points. Although morphological analysis showed unchanged total area of necrosis/regeneration in the diaphragm after training, there was a trend for larger areas of regeneration than necrosis in the diaphragm of mdx-EX compared to mdx-SD mice. However, the levels of Cx39, a protein associated with active regeneration in damaged muscle, were similar in the diaphragm of mdx-EX and mdx-SD mice. Hsp60 significantly decreased at 45 days in the diaphragm, but not in limb muscles, in both trained and sedentary mdx compared to WT mice. In limb muscles, but not in the diaphragm, Hsp70 and NF-kB p65 levels were increased in mdx mice irrespective of training at 30 and 45 days. Therefore, the diaphragm of mdx mice showed little inflammatory and stress responses over time, and appeared hardly affected by mild endurance training. J. Cell. Physiol. 232: 2044-2052, 2017. © 2016 Wiley Periodicals, Inc.
- Published
- 2017
124. Modulation of the TGF-β1-induced epithelial to mesenchymal transition (EMT) mediated by P1 and P2 purine receptors in MDCK cells
- Author
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Michel P. Rathbone, Natale Belluardo, Patricia Giuliani, Valentina Di Liberto, Patrizia Di Iorio, Francesco Caciagli, Daniele F. Condorelli, Mariachiara Zuccarini, Margherita Rossini, Giuseppa Mudò, Marzia Carluccio, Silvana Buccella, Renata Ciccarelli, Zuccarini, M., Giuliani, P., Buccella, S., DI LIBERTO, V., Mudo', G., Belluardo, N., Carluccio, M., Rossini, M., Condorelli, D., Rathbone, M., Caciagli, F., Ciccarelli, R., and Di Iorio, P.
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0301 basic medicine ,MAPK/ERK pathway ,Madin Darby canine kidney cell ,Epithelial-Mesenchymal Transition ,Fibrosi ,Cell ,Transforming growth factor β1 ,Inflammation ,Stimulation ,Biology ,Epithelial to mesenchymal transition ,Fibrosis ,Madin Darby canine kidney cells ,P1/P2 purinergic receptors ,Transforming growth factor β1 ,Molecular Biology ,Cellular and Molecular Neuroscience ,Cell Biology ,Transforming Growth Factor beta1 ,03 medical and health sciences ,0302 clinical medicine ,Dogs ,medicine ,Animals ,Epithelial–mesenchymal transition ,Receptor ,P1/P2 purinergic receptor ,Receptors, Purinergic P2 ,Mesenchymal stem cell ,Receptors, Purinergic P1 ,Cell biology ,Fibronectin ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,Original Article ,medicine.symptom ,Transforming growth factor - Abstract
Epithelial to mesenchymal transition (EMT) occurs during embryogenesis or under pathological conditions such as hypoxia, injury, chronic inflammation, or tissue fibrosis. In renal tubular epithelial cells (MDCK), TGF-β1 induces EMT by reducing or increasing epithelial or mesenchymal marker expression, respectively. In this study, we confirmed that the cAMP analogues, 8-CPT-cAMP or N6-Ph-cAMP, inhibited the TGF-β1-driven overexpression of the mesenchymal markers ZEB-1, Slug, Fibronectin, and α-SMA. Furthermore, we showed that A1, A2A, P2Y1, P2Y11, and P2X7 purine receptor agonists modulated the TGF-β1-induced EMT through the involvement of PKA and/or MAPK/ERK signaling. The stimulation of A2A receptor reduced the overexpression of the EMT-related markers, mainly through the cAMP-dependent PKA pathway, as confirmed by cell pre-treatment with Myr-PKI. Both A1 and P2Y1 receptor stimulation exacerbated the TGF-β1-driven effects, which were reduced by cell pre-treatment with the MAPK inhibitor PD98059, according to the increased ERK1/2 phosphorylation upon receptor activation. The effects induced by P2Y11 receptor activation were oppositely modulated by PKA or MAPK inhibition, in line with the dual nature of the Gs- and Gq-coupled receptor. Differently, P2X7 receptor induced, per se, similar and not additive effects compared to TGF-β1, after prolonged cell exposure to BzATP. These results suggest a putative role of purine receptors as target for anti-fibrotic agents.
- Published
- 2017
125. Guanosine negatively modulates the gastric motor function in mouse
- Author
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Francesca Maiorana, Maria Grazia Zizzo, Antonella Amato, Flavia Mulè, Natale Belluardo, Giuseppa Mudò, Rosa Serio, Zizzo Mg, Mulè F, Amato A, Maiorana F, Mudò G, Belluardo N, and Serio R.
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Male ,medicine.medical_specialty ,Purine nucleoside phosphorylase ,Guanosine ,Adenosine receptor antagonist ,Settore BIO/09 - Fisiologia ,Adenylyl cyclase ,Mice ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Internal medicine ,Cyclic AMP ,medicine ,Animals ,Cyclic adenosine monophosphate ,Molecular Biology ,Dose-Response Relationship, Drug ,Gastric emptying ,Chemistry ,Stomach ,Muscle, Smooth ,Cell Biology ,Adaptation, Physiological ,Adenosine receptor ,Mice, Inbred C57BL ,guanosine , stomach , relaxation ,Endocrinology ,Gastric Emptying ,Original Article ,Gastrointestinal Motility ,Soluble guanylyl cyclase - Abstract
The aim of the present study was to evaluate if guanine-based purines may affect the gastric motor function in mouse. Thus, the influence of guanosine on the gastric emptying rate in vivo was determined and its effects on spontaneous gastric mechanical activity, detected as changes of the intraluminal pressure, were analyzed in vitro before and after different treatments. Gastric gavage of guanosine (1.75-10 mg/kg) delayed the gastric emptying. Guanosine (30 μM-1 mM) induced a concentration-dependent relaxation of isolated stomach, which was not affected by the inhibition of the purine nucleoside phosphorylase enzyme by 4'-deaza-1'-aza-2'-deoxy-1'-(9-methylene)-immucillin-H. The inhibitory response was antagonized by S-(4-nitrobenzyl)-6-thioinosine, a membrane nucleoside transporter inhibitor, but not affected by 9-chloro-2-(2-furanyl)-[1,2,4]triazolo[1,5-c]quinazolin-5-amine, a nonselective adenosine receptor antagonist, or by tetrodotoxin, a blocker of neuronal voltage-dependent Na(+) channels. Moreover, guanosine-induced effects persisted in the presence of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, an inhibitor of soluble guanylyl cyclase or tetraethylammonium, a nonselective potassium channel blocker, but they were progressively reduced by increasing concentrations of 2'5'dideoxyadenosine, an adenylyl cyclase inhibitor. Lastly, the levels of cyclic adenosine monophosphate (cAMP), measured by ELISA, in gastric full thickness preparations were increased by guanosine. In conclusion, our data indicate that, in mouse, guanosine is able to modulate negatively the gastric motor function, reducing gastric emptying and inducing muscular relaxation. The latter is dependent by its cellular uptake and involves adenylyl cyclase activation and increase in cAMP intracellular levels, while it is independent on neural action potentials, adenosine receptors, and K(+) channel activation.
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- 2013
126. Anxiolytic effects of muscarinic acetylcholine receptors agonist oxotremorine in chronically stressed rats and related changes in BDNF and FGF2 levels in the hippocampus and prefrontal cortex
- Author
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Natale Belluardo, Valentina Di Liberto, Monica Frinchi, Maria Fatima Massenti, Carla Cannizzaro, Angela Vitale, Fulvio Plescia, Vincenzo Verdi, Giuseppa Mudò, Di Liberto, V., Frinchi, M., Verdi, V., Vitale, A., Plescia, F., Cannizzaro, C., Massenti, M., Belluardo, N., and Mudò, G.
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0301 basic medicine ,Male ,medicine.medical_specialty ,Elevated plus maze ,medicine.drug_class ,Behavioral test ,Prefrontal Cortex ,Hippocampal formation ,Anxiety ,Muscarinic Agonists ,Anxiolytic ,Hippocampus ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Muscarinic acetylcholine receptor ,medicine ,Oxotremorine ,Muscarinic acetylcholine receptor M4 ,Animals ,Elevated plus maze test ,Rats, Wistar ,Prefrontal cortex ,mAChR ,Chronic restraint stre ,Forced swimming test ,Pharmacology ,Neurons ,Chemistry ,Brain-Derived Neurotrophic Factor ,Cerebral cortex ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Anti-Anxiety Agents ,Fibroblast Growth Factor 2 ,Chronic restraint stress ,Neurotrophins ,Novelty suppressed feeding test ,Neurotrophin ,Neuroscience ,030217 neurology & neurosurgery ,Stress, Psychological ,medicine.drug - Abstract
Rationale: In depressive disorders, one of the mechanisms proposed for antidepressant drugs is the enhancement of synaptic plasticity in the hippocampus and cerebral cortex. Previously, we showed that the muscarinic acetylcholine receptor (mAChR) agonist oxotremorine (Oxo) increases neuronal plasticity in hippocampal neurons via FGFR1 transactivation. Objectives: Here, we aimed to explore (a) whether Oxo exerts anxiolytic effect in the rat model of anxiety-depression-like behavior induced by chronic restraint stress (CRS), and (b) if the anxiolytic effect of Oxo is associated with the modulation of neurotrophic factors, brain-derived neurotrophic factor (BDNF) and fibroblast growth factor-2 (FGF2), and phosphorylated Erk1/2 (p-Erk1/2) levels in the dorsal or ventral hippocampus and in the medial prefrontal cortex. Methods: The rats were randomly divided into four groups: control unstressed, CRS group, CRS group treated with 0.2mg/kg Oxo, and unstressed group treated with Oxo. After 21days of CRS, the groups were treated for 10days with Oxo or saline. The anxiolytic role of Oxo was tested by using the following: forced swimming test, novelty suppressed feeding test, elevated plus maze test, and light/dark box test. The hippocampi and prefrontal cortex were used to evaluate BDNF and FGF2 protein levels and p-Erk1/2 levels. Results: Oxo treatment significantly attenuated anxiety induced by CRS. Moreover, Oxo treatment counteracted the CRS-induced reduction of BDNF and FGF2 levels in the ventral hippocampus and medial prefrontal cerebral cortex Conclusions: The present study showed that Oxo treatment ameliorates the stress-induced anxiety-like behavior and rescues FGF2 and BDNF levels in two brain regions involved in CRS-induced anxiety, ventral hippocampal formation, and medial prefrontal cortex.
- Published
- 2016
127. Parkinson's disease: towards better preclinical models and personalized treatments
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Giuseppa Mudò, Dan Lindholm, Mart Saarma, Ove Eriksson, Johanna Mäkelä, Natale Belluardo, Valentina Di Liberto, Medicum, Department of Biochemistry and Developmental Biology, Institute of Biotechnology, Lindholm, D., Mäkelä, J., Di Liberto, V., Mudo, G., Belluardo, N., Eriksson, O., and Saarma M.
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0301 basic medicine ,Parkinson's disease ,education ,MEDLINE ,Bioinformatics ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,DEFICITS ,Medicine ,Animals ,Humans ,Molecular Biology ,Pharmacology ,business.industry ,Parkinson Disease ,Cell Biology ,medicine.disease ,Molecular medicine ,3. Good health ,MICE ,030104 developmental biology ,Neuroprotective Agents ,Molecular Medicine ,3111 Biomedicine ,business ,030217 neurology & neurosurgery - Abstract
Non
- Published
- 2016
128. Lack of Dystrophin Affects Bronchial Epithelium in mdx Mice
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Giuseppe, Morici, Francesca, Rappa, Francesco, Cappello, Elisabetta, Pace, Andrea, Pace, Giuseppa, Mudò, Grazia, Crescimanno, Natale, Belluardo, Maria R, Bonsignore, Morici, G., Rappa, F., Cappello, F., Pace, E., Pace, A., Mudò, G., Crescimanno, G., Belluardo, N., and Bonsignore, M.
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Apoptosis ,Chaperonin 60 (HSP60) ,Dystrophin ,Goblet cells ,Proliferation ,Clinical Biochemistry ,Cell Biology ,Physiology ,Gene Expression ,Apoptosi ,Bronchi ,Epithelium ,Mice, Inbred C57BL ,Muscular Dystrophy, Duchenne ,Mice, Inbred mdx ,Animals ,Regeneration ,Muscle, Skeletal ,Goblet cell - Abstract
Mild exercise training may positively affect the course of Duchenne Muscular Dystrophy (DMD). Training causes mild bronchial epithelial injury in both humans and mice, but no study assessed the effects of exercise in mdx mice, a well known model of DMD. The airway epithelium was examined in mdx (C57BL/10ScSn-Dmdmdx) mice, and in wild type (WT, C57BL/10ScSc) mice either under sedentary conditions (mdx-SD, WT-SD) or during mild exercise training (mdx-EX, WT-EX). At baseline, and after 30 and 45 days of training (5 d/wk for 6 weeks), epithelial morphology and markers of regeneration, apoptosis, and cellular stress were assessed. The number of goblet cells in bronchial epithelium was much lower in mdx than in WT mice under all conditions. At 30 days, epithelial regeneration (PCNA positive cells) was higher in EX than SD animals in both groups; however, at 45 days, epithelial regeneration decreased in mdx mice irrespective of training, and the percentage of apoptotic (TUNEL positive) cells was higher in mdx-EX than in WT-EX mice. Epithelial expression of HSP60 (marker of stress) progressively decreased, and inversely correlated with epithelial apoptosis (r = -0.66, P = 0.01) only in mdx mice. Lack of dystrophin in mdx mice appears associated with defective epithelial differentiation, and transient epithelial regeneration during mild exercise training. Hence, lack of dystrophin might impair repair in bronchial epithelium, with potential clinical consequences in DMD patients. J. Cell. Physiol. 231: 2218-2223, 2016. © 2016 Wiley Periodicals, Inc.
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- 2016
129. Effects of mild aerobic exercise training on the diaphragm in mdx mice
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FRINCHI, Monica, MORICI, Giuseppe, PITRUZZELLA, Alessandro, BARONE, Rosario, DI LIBERTO, Valentina, PACE, Andrea, Perciavalle, V, BELLUARDO, Natale, CAPPELLO, Francesco, MUDO', Giuseppa, BONSIGNORE, Maria Rosaria, Frinchi, M, Morici, G, Pitruzzella, A, Barone, R, Di Liberto, V, Pace, A, Perciavalle, V, Belluardo, N, Cappello, F, Mudo', G, and Bonsignore, MR
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training, diaphragm, Duchenne Muscolar Distrophy, mdx mice, CX39 protein ,Settore BIO/16 - Anatomia Umana ,Settore CHIM/06 - Chimica Organica ,Settore MED/10 - Malattie Dell'Apparato Respiratorio ,Settore BIO/09 - Fisiologia - Abstract
Mild endurance exercise training positively affects limb skeletal muscle in the mdx mice model of Duchenne Muscular Dystrophy (DMD). However, few and controversial data are available on the effects of mild exercise training on the diaphragm of mdx mice. The diaphragm was examined in mdx and wild type mice either under sedentary conditions (mdx-SD, WT-SD) or during mild exercise training (mdx-EX, WT-EX). At baseline and after 30 and 45 days of training (5 d/wk for 6 weeks), diaphragm muscle morphology and Cx39 protein were assessed. In addition, tissue levels of the chaperonin Hsp60 were measured at the same time points in gastrocnemius, quadriceps and diaphragm in each experimental group. Although morphological analysis showed unchanged total area of necrosis/regeneration in the diaphragm after training, there was a trend for regeneration areas to be larger than necrosis areas. However, the levels of Cx39 protein, a marker associated with active degeneration-regeneration process in damaged muscle were similar in the diaphragm of mdx-EX and mdx-SD mice. The diaphragm, but not limb muscles, of both trained and sedentary mdx mice showed decreased Hsp60 expression at 45 days, suggesting exhaustion of potentially protective mechanisms in the diaphragm similar to previous findings in lung epithelium. Compared to the positive effects of exercise training previously observed in limb skeletal muscles, the diaphragm showed little change after training.
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- 2016
130. PGC-1α: a master gene that is hard to master
- Author
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Johanna Mäkelä, Natale Belluardo, Ove Eriksson, Dan Lindholm, Laura Korhonen, Lindholm, D, Eriksson, O, Mäkelä, J, Belluardo, N, and Korhonen, L
- Subjects
medicine.medical_specialty ,Models, Neurological ,Settore BIO/11 - Biologia Molecolare ,RNA-binding protein ,Biology ,Mitochondrion ,Settore BIO/09 - Fisiologia ,Mice ,Cellular and Molecular Neuroscience ,Heat shock protein ,Internal medicine ,medicine ,Animals ,Homeostasis ,Humans ,Receptor ,Molecular Biology ,Transcription factor ,Heat-Shock Proteins ,Mice, Knockout ,Pharmacology ,PGC-1α, Mitochondria, Dopaminergic neurons , Transgenic animal, Adenovirus, Parkinson’s disease ,Dopaminergic Neurons ,Dopaminergic ,RNA-Binding Proteins ,Parkinson Disease ,Cell Biology ,Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ,Mitochondria ,Endocrinology ,Cell metabolism ,Nerve Degeneration ,Trans-Activators ,Molecular Medicine ,Neuroscience ,Transcription Factors - Abstract
Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) is a transcriptional coactivator that favorably affects mitochondrial function. This concept is supported by an increasing amount of data including studies in PGC-1α gene-deleted mice, suggesting that PGC-1α is a rescue factor capable of boosting cell metabolism and promoting cell survival. However, this view has now been called into question by a recent study showing that adeno-associated virus-mediated PGC-1α overexpression causes overt cell degeneration in dopaminergic neurons. How is this to be understood, and can these seemingly conflicting findings tell us something about the role of PGC-1α in cell stress and in control of neuronal homeostasis?
- Published
- 2012
131. Fibroblast Growth Factor Receptor 1– 5-Hydroxytryptamine 1A Heteroreceptor Complexes and Their Enhancement of Hippocampal Plasticity
- Author
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Valentina Di Liberto, Alexander O. Tarakanov, Dasiel O. Borroto-Escuela, Natale Belluardo, Francisco Ciruela, Mileidys Pérez-Alea, Kjell Fuxe, Manuel Narváez, Giuseppa Mudò, Luigi F. Agnati, Wilber Romero-Fernandez, Borroto-Escuela, DO, Romero-Fernandez, W, Mudó, G, Pérez-Alea, M, Ciruela, F, Tarakanov, AO, Narvaez, M, Di Liberto, V, Agnati, LF, Belluardo, N, and Fuxe, K
- Subjects
Agonist ,medicine.medical_specialty ,Receptor complex ,medicine.drug_class ,Proximity ligation assay ,Biology ,Hippocampal formation ,Transfection ,Heteroreceptor ,Settore BIO/09 - Fisiologia ,Hippocampus ,Rats, Sprague-Dawley ,Growth factor receptor ,Internal medicine ,Fluorescence Resonance Energy Transfer ,medicine ,Animals ,Humans ,Immunoprecipitation ,Receptor, Fibroblast Growth Factor, Type 1 ,Enzyme Inhibitors ,RNA, Small Interfering ,Cells, Cultured ,Biological Psychiatry ,Neurons ,8-Hydroxy-2-(di-n-propylamino)tetralin ,Neuronal Plasticity ,Dose-Response Relationship, Drug ,Fibroblast growth factor receptor 1 ,Computational Biology ,Allosteric modulation, depression, fibroblast growth factor receptor, heteroreceptor, neuronal plasticity, serotonin receptors ,Rats ,Serotonin Receptor Agonists ,Cell biology ,Endocrinology ,Animals, Newborn ,Fibroblast growth factor receptor ,Receptor, Serotonin, 5-HT1A ,Fibroblast Growth Factor 2 ,Peptides ,Signal Transduction - Abstract
Background The hippocampus and its 5-hydroxytryptamine transmission plays an important role in depression related to its involvement in limbic circuit plasticity. Methods The analysis was made with bioluminescence resonance energy transfer, co-immunoprecipitation, in situ proximity ligation assay, binding assay, in cell western and the forced swim test. Results Using bioluminescence resonance energy transfer analysis, fibroblast growth factor receptor 1 (FGFR1)-5-hydroxytryptamine 1A (5-HT1A) receptor complexes have been demonstrated and their specificity and agonist modulation characterized. Their presence based on co-immunoprecipitation and proximity ligation assay has also been indicated in hippocampal cultures and rat dorsal hippocampal formation showing a neuronal location. In vitro assays on extracellular signal-regulated kinases 1 and 2 phosphorylation have shown synergistic increases in signaling on coactivation with fibroblast growth factor 2 (FGF2) and a 5-HT1A agonist, and dependent on the heteroreceptor interface. In vitro and in vivo studies also revealed a 5-HT1A agonist induced phosphorylation of FGFR1 and extracellular signal-regulated kinase 1/2 in rat hippocampus without changing FGF2 levels. Co-activation of the heteroreceptor also resulted in synergistic increases in extensions of PC12 cells and neurite densities and protrusions in primary hippocampal cultures dependent on the receptor interface. The combined acute and repeated intracerebroventricular treatment with FGF2 and 8-OH-DPAT was found to produce evidence of highly significant antidepressant actions in the forced swim test. Conclusions The findings indicate that neurotrophic and antidepressant effects of 5-HT in brain may, in part, be mediated by activation of the 5-HT1A receptor protomer in the hippocampal FGFR1–5-HT1A receptor complex enhancing the FGFR1 signaling.
- Published
- 2012
132. Transgenic expression and activation of PGC-1α protect dopaminergic neurons in the MPTP mouse model of Parkinson’s disease
- Author
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Valentina Di Liberto, Petteri Piepponen, Jose A. Aguirre, Annika Mälkiä, Johanna Mäkelä, Ove Eriksson, Minna Kairisalo, Natale Belluardo, Melania Olivieri, Giuseppa Mudò, Dan Lindholm, Timofey V. Tselykh, Laura Korhonen, Alessandra Bonomo, Mudò, G, Mäkelä, J, Di Liberto, V, Tselykh, TV, Olivieri, M, Piepponen, P, Eriksson, O, Mälkiä, A, Bonomo, A, Kairisalo, M, Aguirre, JA, Korhonen, L, Belluardo, N, and Lindholm, D
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Male ,SOD2 ,Mice, Transgenic ,Substantia nigra ,Mitochondrion ,Biology ,Neuroprotection ,Cell Line ,Mice ,Cellular and Molecular Neuroscience ,chemistry.chemical_compound ,Dopamine ,medicine ,Animals ,Neurotoxin ,Parkinson Disease, Secondary ,Molecular Biology ,PGC-1α, RSV, SIRT1, MPTP, Dopaminergic neurons, Parkinson’s disease ,Pharmacology ,MPTP ,Dopaminergic ,Brain ,Parkinson Disease ,Cell Biology ,Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ,Mitochondria ,Cell biology ,Disease Models, Animal ,Oxidative Stress ,nervous system ,Biochemistry ,chemistry ,1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ,Trans-Activators ,Molecular Medicine ,Female ,Transcription Factors ,medicine.drug - Abstract
Mitochondrial dysfunction and oxidative stress occur in Parkinson’s disease (PD), but little is known about the molecular mechanisms controlling these events. Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) is a transcriptional coactivator that is a master regulator of oxidative stress and mitochondrial metabolism. We show here that transgenic mice overexpressing PGC-1α in dopaminergic neurons are resistant against cell degeneration induced by the neurotoxin MPTP. The increase in neuronal viability was accompanied by elevated levels of mitochondrial antioxidants SOD2 and Trx2 in the substantia nigra of transgenic mice. PGC-1α overexpression also protected against MPTP-induced striatal loss of dopamine, and mitochondria from PGC-1α transgenic mice showed an increased respiratory control ratio compared with wild-type animals. To modulate PGC-1α, we employed the small molecular compound, resveratrol (RSV) that protected dopaminergic neurons against the MPTP-induced cell degeneration almost to the same extent as after PGC-1α overexpression. As studied in vitro, RSV activated PGC-1α in dopaminergic SN4741 cells via the deacetylase SIRT1, and enhanced PGC-1α gene transcription with increases in SOD2 and Trx2. Taken together, the results reveal an important function of PGC-1α in dopaminergic neurons to combat oxidative stress and increase neuronal viability. RSV and other compounds acting via SIRT1/PGC-1α may prove useful as neuroprotective agents in PD and possibly in other neurological disorders.
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- 2011
133. Agonist-induced formation of FGFR1 homodimers and signaling differ among members of the FGF family
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Dasiel O. Borroto-Escuela, Giuseppa Mudò, Natale Belluardo, Kjell Fuxe, Francisco Ciruela, Luigi F. Agnati, Manuel Narváez, Alexander O. Tarakanov, Mileidys Pérez-Alea, Wilber Romero-Fernandez, Romero-Fernandez, W, Borroto-Escuela, DO, Tarakanov, AO, Mudó, G, Narvaez, M, Pérez-Alea, M, Agnati, LF, Ciruela, F, Belluardo, N, and Fuxe, K
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Agonist ,MAPK/ERK pathway ,medicine.drug_class ,Biophysics ,Settore BIO/11 - Biologia Molecolare ,Biology ,Ligands ,Fibroblast growth factor ,Settore BIO/09 - Fisiologia ,Biochemistry ,chemistry.chemical_compound ,Fluorescence Resonance Energy Transfer ,medicine ,Humans ,Receptor, Fibroblast Growth Factor, Type 1 ,Molecular Biology ,Mitogen-Activated Protein Kinase 1 ,Mitogen-Activated Protein Kinase 3 ,Fibroblast growth factor receptor 1 ,HEK 293 cells ,Autophosphorylation ,Cell Biology ,Heparan sulfate ,Fibroblast growth factors, FGFR1, Homodimerization, BRET, MAPK ,Cell biology ,Fibroblast Growth Factors ,stomatognathic diseases ,HEK293 Cells ,chemistry ,Settore BIO/14 - Farmacologia ,Phosphorylation ,Heparitin Sulfate ,Protein Multimerization - Abstract
Fibroblast growth factor receptor 1 (FGFR1) is known to be activated by homodimerization in the presence of both the FGF agonist ligand and heparan sulfate glycosaminoglycan. FGFR1 homodimers in turn trigger a variety of downstream signaling cascades via autophosphorylation of tyrosine residues in the cytoplasmic domain of FGFR1. By means of Bioluminescence Energy Resonance Transfer (BRET) as a sign of FGFR1 homodimerization, we evaluated in HEK293T cells the effects of all known FGF agonist ligands on homodimer formation. A significant correlation between BRET(2) signaling and ERK1/2 phosphorylation was observed, leading to a further characterization of the binding and signaling properties of the FGF subfamilies. FGF agonist ligand-FGFR1 binding interactions appear as the main mechanism for the control of FGFR1 homodimerization and MAPK signaling which demonstrated a high correlation. The bioinformatic analysis demonstrates the interface of the two pro-triplets SSS (Ser-Ser-Ser) and YGS (Tyr-Gly-Ser) located in the extracellular and intracellular domain of the FGFR1. These pro-triplets are postulated participate in the FGFR1 homodimerization interface interaction. The findings also reveal that FGF agonist ligands within the same subfamily of the FGF gene family produced similar increases in FGFR1 homodimer formation and MAPK signaling. Thus, the evolutionary relationship within this gene family appears to have a distinct functional relevance.
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- 2011
134. FGF-2/FGFR1 neurotrophic system expression level and its basal activation do not account for the age-dependent decline of precursor cell proliferation in the subventricular zone of rat brain
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Monica Frinchi, Kjell Fuxe, Melania Olivieri, Giuseppa Mudò, Valentina Di Liberto, Natale Belluardo, Frinchi, M, Di Liberto, V, Olivieri, M, Fuxe, K, Belluardo, N, and Mudò, G
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Male ,Aging ,medicine.medical_specialty ,Subventricular zone ,Neurogenesis ,Receptor expression ,FGF-2 ,Fibroblast growth factor ,Settore BIO/09 - Fisiologia ,Cerebral Ventricles ,FGFR1 ,Neuronal precursor cells ,Growth factor receptor ,Neurotrophic factors ,Internal medicine ,Precursor cell ,medicine ,Animals ,RNA, Messenger ,Receptor, Fibroblast Growth Factor, Type 1 ,Phosphorylation ,Rats, Wistar ,Molecular Biology ,Cell Proliferation ,Mitogen-Activated Protein Kinase 3 ,biology ,Phospholipase C gamma ,General Neuroscience ,Brain ,Neuronal precursor cell ,Rats ,Adult Stem Cells ,medicine.anatomical_structure ,Endocrinology ,Bromodeoxyuridine ,Gene Expression Regulation ,biology.protein ,Fibroblast Growth Factor 1 ,Neurogenesi ,Fibroblast Growth Factor 2 ,Neurology (clinical) ,Developmental Biology ,Neurotrophin - Abstract
It is largely accepted that neurogenesis in the adult brain decreases with age and reduced levels of local neurotrophic support is speculated to be a contributing factor. Among neurotrophic factors involved on neurogenesis, we focused our attention on the neurotrophic system fibroblast growth factor-2 (FGF-2) and its receptor FGFR1, a potent modulator of precursor cell proliferation. In the present work, we aimed to analyse if potential age-dependent changes of the FGF-2/FGFR1 neurotrophic system may give account for the age-dependent decline of precursor cell proliferation in the neurogenic region of the subventricular zone (SVZ) in the rat brain. Using in situ hybridization and western blotting procedures we examined FGF-2 and FGFR1 expression levels in the SVZ of 20-month-old rats as compared to young adult 3-month-old rats. The results showed that during aging the FGF-2 and its receptor expression levels, both as mRNA and protein, were unchanged in the SVZ. The levels of phosphorylated FGFR1 form did not show significant variations suggesting that also the level of receptor activation does not change during aging. No changes were also observed in the phosphorylation of two FGFR1 related proteins involved in intracellular signaling, the canonical extracellular signal-regulated kinase Erk1/2 and the phospholipase-Cγ1. Additionally, we could show that also the proliferation rate of stem cells does not change during aging. Taken together, our results show that FGF-2/FGFR1 neurotrophic system expression level and its basal activation do not account for the age-dependent decline of precursor cell proliferation in the rat brain.
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- 2010
135. Small airways in sedentary and endurance-trained dystrophic (mdx) mice
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RAPPA, Francesca, BONSIGNORE, Maria Rosaria, FRINCHI, Monica, MUDO', Giuseppa, BELLUARDO, Natale, CAPPELLO, Francesco, MORICI, Giuseppe, Rappa, F, Bonsignore, M, Frinchi, M, Mudò, G, Belluardo,N, Cappello, F, and Morici, G
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Settore BIO/16 - Anatomia Umana ,Settore MED/10 - Malattie Dell'Apparato Respiratorio ,Smal airways, dystrophic mice ,Settore BIO/09 - Fisiologia - Abstract
The effects of mild endurance exercise training on the small airways in mdx mice are unknown. We compared epithelial thickness and turnover, apoptosis, and stress marker expression in small airways of mdx mice and wild-type (WT) controls, at rest and during exercise training. Mdx and WT mice were randomly assigned to sedentary (mdx-S, n=17; WT-S, n=19) or trained (mdx- EX, n=14; WT-EX, n=16) groups. Low-intensity endurance training (running on a wheel) was done 5 d/wk for 6 wk at progressively increasing speed (rpm from 16 to 24) and time (15 min to 1 h). Lungs were processed for light microscopy and periodic acid Schiff (PAS) staining. Hsp60 and PCNA were quantified by immunohistochemistry. Apoptosis was assessed by TUNEL. Bronchial epithelial thickness decreased over time in WT mice irrespective of training (linear regression for time trends: WT-S: R2=0.43, r= -0.65; WT-EX: R2=0.68, r= -0.82, p
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- 2015
136. Enhancement of the FGFR1 signaling in the FGFR1-5-HT1A heteroreceptor complex in midbrain raphe 5-HT neuron systems. Relevance for neuroplasticity and depression
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Manuel Narváez, Mileidys Pérez-Alea, Giuseppa Mudò, Kjell Fuxe, Francisco Ciruela, Dasiel O. Borroto-Escuela, Alexander O. Tarakanov, Antonio Jiménez-Beristain, Luigi F. Agnati, Natale Belluardo, Borroto-Escuela, D., Pérez-Alea, M., Narvaez, M., Tarakanov, A., Mudó, G., Jiménez-Beristain, A., Agnati, L., Ciruela, F., Belluardo, N., and Fuxe, K.
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Agonist ,Serotonin ,medicine.medical_specialty ,medicine.drug_class ,Cellular differentiation ,Biophysics ,Heteroreceptor complex ,Biology ,Heteroreceptor ,Biochemistry ,Settore BIO/09 - Fisiologia ,Cell Line ,Midbrain ,Dorsal raphe nucleus ,Mesencephalon ,Internal medicine ,medicine ,Animals ,Serotonin 5-HT1A receptor ,Receptor, Fibroblast Growth Factor, Type 1 ,Protein Interaction Maps ,Phosphorylation ,Extracellular Signal-Regulated MAP Kinases ,Molecular Biology ,Neurons ,8-Hydroxy-2-(di-n-propylamino)tetralin ,Neuronal Plasticity ,Raphe ,Depression ,Animal ,Extracellular Signal-Regulated MAP Kinase ,Cell Biology ,Serotonin 5-HT1 Receptor Agonists ,Neuron ,Fibroblast growth factor receptor ,Rats ,Endocrinology ,medicine.anatomical_structure ,nervous system ,Receptor, Serotonin, 5-HT1A ,Autoreceptor ,Rat ,Fibroblast Growth Factor 2 ,Serotonin 5-HT1 Receptor Agonist ,Dimerization ,Neuroscience ,Neuronal plasticity - Abstract
New findings show existence of FGFR1-5-HT1A heteroreceptor complexes in 5-HT nerve cells of the dorsal and median raphe nuclei of the rat midbrain and hippocampus. Synergistic receptor-receptor interactions in these receptor complexes indicated their enhancing role in hippocampal plasticity. The existence of FGFR1-5-HT1A heteroreceptor complexes also in midbrain raphe 5-HT nerve cells open up the possibility that antidepressant drugs by increasing extracellular 5-HT levels can cause an activation of the FGF-2/FGFR1 mechanism in these nerve cells as well. Therefore, the agonist modulation of the FGFR1-5-HT1A heteroreceptor complexes and their specific role is now determined in rat medullary raphe RN33B cells and in the caudal midline raphe area of the midbrain rich in 5-HT nerve cells. The combined i.c.v. treatment with FGF-2 and the 5-HT1A agonist 8-OHDPAT synergistically increased FGFR1 and ERK1/2 phosphorylation in the raphe midline area of the midbrain and in the RN33B cells. Cotreatment with FGF2 and the 5-HT1A agonist induced RN33B cell differentiation as seen from development of an increased number and length of extensions per cell and their increased 5-HT immunoreactivity. These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TMV but not by TMII of the 5-HT1A receptor. Taken together, the 5-HT1A autoreceptors by being part of a FGFR1-5-HT1A heteroreceptor complex in the midbrain raphe 5-HT nerve cells appears to have also a trophic role in the central 5-HT neuron systems besides playing a key role in reducing the firing of these neurons.
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- 2015
137. Reduction of mdx mouse muscle degeneration by low-intensity endurance exercise: a proteomic analysis in quadriceps muscle of exercised versus sedentary mdx mice
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Odessa Schillaci, Valentina Di Liberto, Vincenzo Perciavalle, Marco Giallombardo, Natale Belluardo, Simona Fontana, Monica Frinchi, Riccardo Alessandro, Giacomo De Leo, Giuseppa Mudò, Giuseppe Morici, Fontana, S., Schillaci, O., Frinchi, M., Giallombardo, M., Morici, G., Di Liberto, V., Alessandro, R., De Leo, G., Perciavalle, V., Belluardo, N., and Mudo', G.
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Male ,Proteomics ,muscular dystrophy ,mdx mouse ,medicine.medical_specialty ,carbonic anhydrase, exercise, mdx, muscle oxidative stress, muscle proteomic, muscular dystrophy ,Blotting, Western ,carbonic anhydrase ,Biophysics ,Muscle Proteins ,medicine.disease_cause ,Biochemistry ,Quadriceps Muscle ,muscle proteomic ,Superoxide dismutase ,Western blot ,Endurance training ,Internal medicine ,medicine ,Animals ,oxidative stress ,Electrophoresis, Gel, Two-Dimensional ,Muscular dystrophy ,Molecular Biology ,Original Paper ,exercise ,biology ,medicine.diagnostic_test ,Superoxide Dismutase ,business.industry ,Reproducibility of Results ,Skeletal muscle ,Cell Biology ,medicine.disease ,Original Papers ,Carbonic Anhydrase III ,Mice, Inbred C57BL ,Muscular Dystrophy, Duchenne ,medicine.anatomical_structure ,Endocrinology ,X-linked muscular dystrophy (mdx) ,Mice, Inbred mdx ,Physical Endurance ,biology.protein ,Carbonic anhydrase 3 ,business ,muscle oxidative stress ,Oxidative stress - Abstract
By proteomic analysis we found an up-regulation of four carbonic anhydrase-3 (CA3) isoforms and a down-regulation of superoxide dismutase [Cu-Zn] (SODC) in quadriceps of sedentary X-linked muscular dystrophy (mdx) mice as compared with wild–type (WT) mice and the levels were significantly restored to WT values following low-intensity endurance exercise., In our recent study was shown a significant recovery of damaged skeletal muscle of mice with X-linked muscular dystrophy (mdx) following low-intensity endurance exercise, probably by reducing the degeneration of dystrophic muscle. Consequently, in the present work, we aimed to identify proteins involved in the observed reduction in degenerating fibres. To this end, we used proteomic analysis to evaluate changes in the protein profile of quadriceps dystrophic muscles of exercised compared with sedentary mdx mice. Four protein spots were found to be significantly changed and were identified as three isoforms of carbonic anhydrase 3 (CA3) and superoxide dismutase [Cu-Zn] (SODC). Protein levels of CA3 isoforms were significantly up-regulated in quadriceps of sedentary mdx mice and were completely restored to wild–type (WT) mice values, both sedentary and exercised, in quadriceps of exercised mdx mice. Protein levels of SODC were down-regulated in quadriceps of sedentary mdx mice and were significantly restored to WT mice values, both sedentary and exercised, in quadriceps of exercised mdx mice. Western blot data were in agreement with those obtained using proteomic analysis and revealed the presence of one more CA3 isoform that was significantly changed. Based on data found in the present study, it seems that low-intensity endurance exercise may in part contribute to reduce cell degeneration process in mdx muscles, by counteracting oxidative stress.
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- 2015
138. Nicotinic receptor agonists as neuroprotective/neurotrophic drugs. Progress in molecular mechanisms
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Giuseppa Mudò, Kjell Fuxe, Natale Belluardo, Mudo', G, Belluardo, N, and Fuxe, K
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neuroplasticity ,desensitization ,FGF-2 ,nAChR ,Receptors, Nicotinic ,Biology ,Pharmacology ,Settore BIO/09 - Fisiologia ,Neuroprotection ,Nicotine ,Neurotrophic factors ,medicine ,Animals ,Humans ,nicotinic agonist ,Nerve Growth Factors ,Nicotinic Agonists ,neurotrophic factor ,Biological Psychiatry ,Neuronal Plasticity ,nicotinic agonists ,neurotrophic factors ,neuroprotection ,neurotrophism ,addiction ,Brain ,Neurodegenerative Diseases ,Tobacco Use Disorder ,Psychiatry and Mental health ,Neuroprotective Agents ,Nerve growth factor ,Nicotinic agonist ,Neurology ,Synaptic plasticity ,biology.protein ,Neurology (clinical) ,Signal transduction ,Neuroscience ,Signal Transduction ,medicine.drug ,Neurotrophin - Abstract
In the present work we reviewed recent advances concerning neuroprotective/neurotrophic effects of acute or chronic nicotine exposure, and the signalling pathways mediating these effects, including mechanisms implicated in nicotine addiction and nAChR desensitization. Experimental and clinical data largely indicate long-lasting effects of nicotine and nicotinic agonists that imply a neuroprotective/neurotrophic role of nAChR activation, involving mainly alpha 7 and alpha 4 beta 2 nAChR subtypes, as evidenced using selective nAChR agonists. Compounds interacting with neuronal nAChRs have the potential to be neuroprotective and treatment with nAChR agonists elicits long-lasting neurotrophic effects, e.g. improvement of cognitive performance in a variety of behavioural tests in rats, monkeys and humans. Nicotine addiction, which is mediated by interaction with nACh receptors, is believed to involve the modification of signalling cascades that modulate synaptic plasticity and gene expression. Desensitization, in addition to protecting cells from uncontrolled excitation, is recently considered as a form of signal plasticity. nAChR can generate these longe-lasting effects by elaboration of complex intracellular signals that mediate medium to long-term events crucial for neuronal maintenance, survival and regeneration. Although a comprehensive survey of the gene-based molecular mechanisms that underlie nicotine effects has yet not been performed a growing amount of data is beginning to improve our understanding of signalling mechanisms that lead to neurotrophic/neuroprotective responses. Evidence for an involvement of the fibroblast growth factor-2 gene in nAChR mechanisms mediating neuronal survival, trophism and plasticity has been obtained. However, more work is needed to establish the mechanisms involved in the effects of nicotinic receptor subtype activation from cognition-enhancing and neurotrophic effects to smoking behaviour and to determine more precisely the therapeutic objectives in potential nicotinic drug treatments of neurodegenerative diseases. In the present work we reviewed recent advances concerning neuroprotective/neurotrophic effects of acute or chronic nicotine exposure, and the signalling pathways mediating these effects, including mechanisms implicated in nicotine addiction and nAChR desensitization. Experimental and clinical data largely indicate long-lasting effects of nicotine and nicotinic agonists that imply a neuroprotective/neurotrophic role of nAChR activation, involving mainly alpha 7 and alpha 4 beta 2 nAChR subtypes, as evidenced using selective nAChR agonists. Compounds interacting with neuronal nAChRs have the potential to be neuroprotective and treatment with nAChR agonists elicits long-lasting neurotrophic effects, e.g. improvement of cognitive performance in a variety of behavioural tests in rats, monkeys and humans. Nicotine addiction, which is mediated by interaction with nACh receptors, is believed to involve the modification of signalling cascades that modulate synaptic plasticity and gene expression. Desensitization, in addition to protecting cells from uncontrolled excitation, is recently considered as a form of signal plasticity. nAChR can generate these longe-lasting effects by elaboration of complex intracellular signals that mediate medium to long-term events crucial for neuronal maintenance, survival and regeneration. Although a comprehensive survey of the gene-based molecular mechanisms that underlie nicotine effects has yet not been performed a growing amount of data is beginning to improve our understanding of signalling mechanisms that lead to neurotrophic/neuroprotective responses. Evidence for an involvement of the fibroblast growth factor-2 gene in nAChR mechanisms mediating neuronal survival, trophism and plasticity has been obtained. However, more work is needed to establish the mechanisms involved in the effects of nicotinic receptor subtype activation from cognition-enhancing and neurotrophic effects to smoking behaviour and to determine more precisely the therapeutic objectives in potential nicotinic drug treatments of neurodegenerative diseases.
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- 2006
139. Evidence for the existence of FGFR1-5-HT1A heteroreceptor complexes in the midbrain raphe 5-HT system
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Antonio Jiménez-Beristain, Francisco Ciruela, Luigi F. Agnati, Kjell Fuxe, Dasiel O. Borroto-Escuela, Natale Belluardo, Giuseppa Mudò, Manuel Narváez, Mileidys Pérez-Alea, Alexander O. Tarakanov, Borroto-Escuela, D, Narvaez, M, Pérez-Alea, M, Tarakanov, A, Jiménez-Beristain, A, Mudó, G, Agnati, L, Ciruela, F, Belluardo, N, and Fuxe, K
- Subjects
Male ,medicine.medical_specialty ,Serotonin ,G-protein-coupled receptor ,Receptor tyrosine kinase ,Biophysics ,Heteroreceptor complex ,Proximity ligation assay ,Biology ,Heteroreceptor ,Biochemistry ,Midbrain ,Rats, Sprague-Dawley ,G-protein-coupled receptors ,Receptor tyrosine kinases ,Fibroblast growth factor receptor 1 ,Serotonin receptors ,Dimerization ,Internal medicine ,medicine ,Fluorescence Resonance Energy Transfer ,Animals ,Humans ,Receptor, Fibroblast Growth Factor, Type 1 ,Serotonin receptor ,Molecular Biology ,5-HT receptor ,Neurons ,8-Hydroxy-2-(di-n-propylamino)tetralin ,Raphe ,Midbrain Raphe Nuclei ,Cell Biology ,Cell biology ,Rats ,medicine.anatomical_structure ,Endocrinology ,HEK293 Cells ,nervous system ,Gene Expression Regulation ,Receptor, Serotonin, 5-HT1A ,Autoreceptor ,Fibroblast Growth Factor 2 ,Neuron ,Raphe nuclei ,Peptides ,Protein Binding - Abstract
The ascending midbrain 5-HT neurons known to contain 5-HT1A autoreceptors may be dysregulated in depression due to a reduced trophic support. With in situ proximity ligation assay (PLA) and supported by co-location of the FGFR1 and 5-HT1A immunoreactivities in midbrain raphe 5-HT cells, evidence for the existence of FGFR1–5-HT1A heteroreceptor complexes were obtained in the dorsal and median raphe nuclei of the Sprague–Dawley rat. Their existence in the rat medullary raphe RN33B cell cultures was also established. After combined FGF-2 and 8-OH-DPAT treatment, a marked and significant increase in PLA positive clusters was found in the RN33B cells. Similar results were reached upon coactivation by agonists in HEK293T cells using the Fluorescent Resonance Energy Transfer (FRET) technique resulting in increased FRETmax and reduced FRET50 values. The heteroreceptor complex formation was dependent on TMV of the 5-HT1A receptor since it was blocked by incubation with TMV but not with TMII. Taken together, the 5-HT1A autoreceptors by being recruited into a FGFR1–5-HT1A heteroreceptor complex in the midbrain raphe 5-HT nerve cells may develop a novel function, namely a trophic role in many midbrain 5-HT neuron systems originating from the dorsal and medianus raphe nuclei.
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- 2014
140. Transcription factor gene expression profiling after acute intermittent nicotine treatment in the rat cerebral cortex
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Kjell Fuxe, Giuseppe Amato, Giuseppa Mudò, Wolfgang H. Sommer, P.-A. Olsson, Natale Belluardo, BELLUARDO N, OLSSON PA, MUDO' G, SOMMER WH, AMATO G, and FUXE K
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Male ,Nicotine ,Gene Expression ,Biology ,Neuroprotection ,Cerebral Cortex/drug effect ,Neurotrophic factors ,medicine ,Animals ,Nicotinic Agonists ,Rats, Wistar ,In Situ Hybridization ,Oligonucleotide Array Sequence Analysis ,Cerebral Cortex ,Microarray analysis techniques ,Gene Expression Profiling ,General Neuroscience ,Rats ,Gene expression profiling ,Nicotinic agonist ,Nicotine/pharmacology Nicotinic Agonists ,biology.protein ,Gene Expression/drug effect ,Immediate early gene ,Neuroscience ,Transcription Factors ,medicine.drug ,Neurotrophin - Abstract
Several studies in different in vitro and in vivo models have demonstrated neuroprotective effects of nicotinic receptor agonists and indirect trophic actions of nicotine on brain are suggested from observations describing nicotine as a cognitive enhancer by increasing vigilance and improving learning and memory. While an increasing number of studies have given evidence of neuroprotective and neurotrophic effects of nicotine treatment, the molecular mechanism mediating the neurotrophic effects of nicotine are not fully understood. Previously in an analysis of several neurotrophic factors as possible mediators of nicotine-induced neuroprotection and/or neurotrophic effects we could reveal that an acute intermittent nicotine treatment increases fibroblast growth factor-2 mRNA and protein in several brain regions of rat brain. Even if other studies have demonstrated in different paradigms that nicotine administration modulates expression level of a variety of genes, there is still a lack of indication which candidate genes, involved in neuroprotective responses are modulated by nicotine. In the present work we have used a microarray assay to further find and characterize new genes responsive to acute intermittent nicotine treatment and linked to neuroprotection. Therefore, we used Rat Genome U34A Affymetrix GeneChip arrays containing about 8800 probe sets to characterize transcriptional responses in the rat parietal cortex after acute intermittent nicotine treatment. We focused our attention to expression of transcription factors and several of them were up- or down-regulated by nicotine, among these Nr4a1 (Nurr77), Egr-1 and Egr-2. In situ hybridization was used to corroborate the microarray data and to reveal further spatial and temporal patterns of these nicotine induced genes. Taken together the present results identified several novel candidate genes modified by acute intermittent nicotine exposure and as such potentially involved in neuroprotective-neurotrophic actions.
- Published
- 2005
141. No effects of low-intensity endurance exercise on muscle necrosis in the diaphragm of mdx mice
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FRINCHI, Monica, MUDO', Giuseppa, RAPPA, Francesca, BONSIGNORE, Maria Rosaria, CAPPELLO, Francesco, BELLUARDO, Natale, PACE, Andrea, MORICI, Giuseppe, Frinchi, M, Mudò, G, Rappa, F, Bonsignore, MR, Cappello,F, Belluardo, N, Pace, A, and Morici, G
- Subjects
training ,Settore BIO/16 - Anatomia Umana ,Cx39 ,Duchenne muscolar dystrophy ,Settore MED/10 - Malattie Dell'Apparato Respiratorio ,Settore BIO/09 - Fisiologia - Abstract
Duchenne muscular dystrophy (DMD) is characterized by progressive skeletal muscle weakness. We have previously shown that low-intensity endurance training prevented muscle damage (Frinchi et al, Int J Sports Med 2014). Since the effects of low-intensity endurance training on the the diaphragm in the mdx mouse model are unknown, in the same animals we investigated Cx39 protein levels (Western blotting) in homogenates of the diaphragm before and after training. Mdx and wild-type (WT) mice were randomly assigned to sedentary (mdx-S, n=17; WT-S, n=19) or trained (mdx-EX, n=14; WT-EX, n=16) groups. Low-intensity endurance training (running on a wheel) was done 5 days/week for 6 weeks at progressively increasing time (15 min to 1 h) and speed (rpm from 16 to 24, distance covered during training sessions from 48 to 288 m). Compared to our previous analysis of skeletal muscles changes in gastrocnemius and quadriceps, showing decreased muscle damage in trained vs sedentary mdx mice, analysis of protein level of Cx39 showed similar values in diaphragm homogenates from sedentary and trained mdx mice. These preliminary data suggest that prevention of muscle necrosis after mild training does not occur in the diaphragm. As a speculation, continuous work of diaphragm vs intermittent work of skeletal muscle might at least partly account for the different results obtained in respiratory and locomotor muscle.
- Published
- 2014
142. Fibroblast growth factor-21 enhances mitochondrial functions and increases the activity of PGC-1α in human dopaminergic neurons via Sirtuin-1
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Hai Thi Do, Timofey V. Tselykh, Francesca Maiorana, Natale Belluardo, Ove Eriksson, Giuseppa Mudò, Johanna Mäkelä, Dan Lindholm, Laura Korhonen, Medicum, Department of Biochemistry and Developmental Biology, Mäkelä, J, Tselykh, TV, Maiorana, F, Eriksson, O, Do, HT, Mudò, G, Korhonen L, Belluardo, N, and Lindholm, D
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FGF21 ,education ,Regulator ,Nicotinamide phosphoribosyltransferase ,PGC-1α ,SIRT1 ,Dopaminergic neurons ,Mitochondria ,Parkinson’s disease ,Mitochondrion ,Bioinformatics ,Settore BIO/09 - Fisiologia ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Dopaminergic Cell ,030304 developmental biology ,Dopaminergic neuron ,0303 health sciences ,Multidisciplinary ,biology ,Sirtuin 1 ,Research ,Dopaminergic ,Cell biology ,chemistry ,biology.protein ,3111 Biomedicine ,NAD+ kinase ,030217 neurology & neurosurgery - Abstract
Mitochondrial dysfunctions accompany several neurodegenerative disorders and contribute to disease pathogenesis among others in Parkinson’s disease (PD). Peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) is a major regulator of mitochondrial functions and biogenesis, and was suggested as a therapeutic target in PD. PGC-1α is regulated by both transcriptional and posttranslational events involving also the action of growth factors. Fibroblast growth factor-21 (FGF21) is a regulator of glucose and fatty acid metabolism in the body but little is known about its action in the brain. We show here that FGF21 increased the levels and activity of PGC-1α and elevated mitochondrial antioxidants in human dopaminergic cells in culture. The activation of PGC-1α by FGF21 occurred via the NAD+-dependent deacetylase Sirtuin-1 (SIRT1) subsequent to an increase in the enzyme, nicotinamide phosphoribosyltransferase (Nampt). FGF21 also enhanced mitochondrial respiratory capacity in human dopaminergic neurons as shown in real-time analyses of living cells. FGF21 is present in the brain including midbrain and is expressed by glial cells in culture. These results show that FGF21 activates PGC-1α and increases mitochondrial efficacy in human dopaminergic neurons suggesting that FGF21 could potentially play a role in dopaminergic neuron viability and in PD.
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- 2014
143. Study of molecular mechanism involved in neuronal plasticity induced by magnetic stimulation in cultured hippocampal neurons
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DI LIBERTO, Valentina, COSENTINO, Giuseppe, Frinchi, M, BRIGHINA, Filippo, Woods, K, FIERRO, Brigida, BELLUARDO, Natale, MUDO', Giuseppa, Di Liberto, V, Cosentino, G, Frinchi, M, Brighina, F, Woods, K, Fierro, B, Belluardo, N, and Mudò, G
- Subjects
Magnetic stimulation, neuronal plasticity, neurotrophic factors - Abstract
Although a large number of investigations have shown that transcranial magnetic stimulation, a non-invasive method of brain stimulation with minimal side effects, is able to induce neuronal synaptic plastic change, very few studies have examined the molecular mechanisms of magnetic stimulation involved in synaptic plasticity. Since it is well known that neurotrophins and their receptors regulate synaptic strength and thereby mediate plasticity, in this study we have investigated the effects of low-frequency (1 Hz) magnetic stimulation, at different intensities, on the activation of neurotrophic factors receptors and relative intracellular pathways in primary cultures of hippocampal neurons. The results showed that one single exposure to magnetic stimulation, low-frequency and 1.55 tesla intensity, activates Glial cell-derived neurotrophic factor receptor (RET), Brain-derived neurotrophic factor receptor (TrkB), Fibroblasts growth factor 2 receptor 1 (FGFR1) and PI3K/Akt pathway in primary cultures of hippocampal neurons after only a short time (5 minutes). These data may explain, at least in part, the mechanism through which magnetic stimulation enhances synaptic plasticity. Our current studies are characterizing the mechanism of neurotrophic factor receptor activation following magnetic stimulation, including the role of neurotransmitters release.
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- 2014
144. FGFR1-5-HT1A receptor heterocomplexes: relevance for neuroplasticity and depression
- Author
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Fuxe, K, Romero Fernandez, W, MUDO', Giuseppa, Pérez Alea, M, Tarakanov, AO, Narvaez, M, Agnati, LF, BELLUARDO, Natale, Borroto Escuela, DO, Fuxe, K, Romero-Fernandez, W, Mudó, G, Pérez-Alea, M, Tarakanov, AO, Narvaez, M, Agnati, LF, Belluardo, N, and Borroto-Escuela, DO
- Subjects
Antidepressants: basic, Depression: basic, Receptors ,Settore BIO/09 - Fisiologia - Published
- 2014
145. Quadriceps muscle proteomic profiling of exercised versus sedentary mdx mice
- Author
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Frinchi, M, FONTANA, Simona, Giallombardo, Marco, SCHILLACI, Odessa, MORICI, Giuseppe, DI LIBERTO, Valentina, ALESSANDRO, Riccardo, BELLUARDO, Natale, MUDO', Giuseppa, Frinchi, M, Fontana, S, Giallombardo, M, Schillaci, O, Morici, G, Di Liberto, V, Alessandro, R, Belluardo, N, and Mudò, G
- Subjects
muscular dystrophy ,Proteomic ,mdx - Published
- 2014
146. Recovery of Damaged Skeletal Muscle in mdx Mice through Low-intensity Endurance Exercise
- Author
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Natale Belluardo, Giuseppa Mudò, M. Coco, Attilio Licciardi, Monica Frinchi, Vincenzo Perciavalle, Filippo Macaluso, Giuseppe Morici, Frinchi,M, Macaluso,F, Licciardi,A, Perciavalle,V, Coco,M, Belluardo,N, Morici,G, and Mudò,G
- Subjects
Male ,musculoskeletal diseases ,medicine.medical_specialty ,Blotting, Western ,Connexin ,Physical Therapy, Sports Therapy and Rehabilitation ,Degeneration (medical) ,Settore BIO/09 - Fisiologia ,Connexins ,Mice ,Random Allocation ,dystrophic muscle, muscle regeneration, muscle injury, connexin, connexin 39, muscle strength ,Endurance training ,Physical Conditioning, Animal ,Internal medicine ,medicine ,Animals ,Regeneration ,Orthopedics and Sports Medicine ,Muscle Strength ,Muscular dystrophy ,Muscle, Skeletal ,biology ,Muscle fatigue ,business.industry ,Skeletal muscle ,Anatomy ,Muscular Dystrophy, Animal ,medicine.disease ,Exercise Therapy ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Endocrinology ,Muscle Fatigue ,Mice, Inbred mdx ,Physical Endurance ,biology.protein ,medicine.symptom ,Dystrophin ,business ,Biomarkers ,Muscle Contraction ,Muscle contraction - Abstract
The lack of dystrophin in mdx mice leads to cycles of muscle degeneration and regeneration processes. Various strategies have been proposed in order to reduce the muscle-wasting component of muscular dystrophy, including implementation of an exercise programme. The aim of this study was to examine how low-intensity endurance exercise affects the degeneration-regeneration process in dystrophic muscle of male mdx mice. Mice were subjected to low-intensity endurance exercise by running on a motorized Rota-Rod for 5 days/week for 6 weeks. Histomorphological analysis showed a signifi cant reduction of measured inflammatory-necrotic areas in both gastrocnemius and quadriceps muscle of exercised mdx mice as compared to matched sedentary mdx mice. The degenerative-regenerative process was also evaluated by examining the protein levels of connexin 39 (Cx39), a specifi c gene expressed in injured muscles. Cx39 was not detected in sedentary wild type mice, whereas it was found markedly increased in sedentary mdx mice, revealing active muscle degeneration-regeneration process. These Cx39 protein levels were signifi cantly reduced in muscles of mdx mice exercised for 30 and 40 days, revealing together with histomorphological analysis a strong reduction of degeneration process in mice subjected to low-intensity endurance exercise. Muscles of exercised mdx mice did not show significant changes in force and fatigue resistance as compared to sedentary mdx mice. Overall in this study we found that specifi c lowintensity endurance exercise induces a beneficial effect probably by reducing the degeneration of dystrophic muscle.
- Published
- 2013
147. Distribution and Function of Gap Junction Coupling in Cortical GABAergic Neurons
- Author
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Vincenza Barresi, Daniele F. Condorelli, Natale Belluardo, Giuseppa Mudò, Ekrem Dere, Condorelli, DF, Mudò, G, Barresi, V, and Belluardo, N
- Subjects
genetic structures ,Interneuron ,GAP Junction, GABAergic neurons ,musculoskeletal, neural, and ocular physiology ,Immunoelectron microscopy ,Gap junction ,Hippocampus ,Settore BIO/11 - Biologia Molecolare ,Biology ,Settore BIO/09 - Fisiologia ,medicine.anatomical_structure ,Electrical Synapses ,nervous system ,Cerebral cortex ,Settore BIO/10 - Biochimica ,Synaptic plasticity ,medicine ,GABAergic ,Neuroscience - Abstract
Although gap junctions have been observed in GABAergic interneurons of several brain regions, this chapter focuses on the distribution and functions of gap junctions and connexins in inhibitory interneurons of the cerebral cortex and hippocampus. Evidence for interconnections mediated by electrical synapses is reported for at least eight cerebral cortex interneuron types, classified on the basis of morphology, electrophysiology and molecular markers. The main differences in the organization of these interneuronal networks are summarized in terms of homologous and heterologous electrical coupling and mutual chemical inhibition. The role of connexin36 (Cx36) in forming neuronal electrical synapses in interneurons is reviewed by analyzing data derived by techniques such as immunoelectron microscopy, in situ hybridization, immunohistochemistry, reporter gene expression in transgenic mice, and functional and behavioral studies in knockout mice. Possible functional roles of Cx36-based gap junctions in GABAergic interneuronal networks are examined, from the molecular and cellular level to generation of oscillatory activities and synaptic plasticity.
- Published
- 2013
148. Connexin36 (Cx36) expression and protein detection in the mouse carotid body and myenteric plexus
- Author
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Giuseppa Mudò, Monica Frinchi, Francesca D’Antoni, Sada Lakshmi Turimella, Natale Belluardo, Valentina Di Liberto, Martin Theis, Frinchi, M, Di Liberto, V, Turimella, S, D'Antoni, F, Theis, M, Belluardo, N, and Mudò, G
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Histology ,Mouse ,genetic structures ,Myenteric plexus ,Blotting, Western ,Ileum ,Connexin ,Biology ,Settore BIO/09 - Fisiologia ,Connexins ,Mice ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Animals ,Gap junctions ,030304 developmental biology ,Mice, Knockout ,0303 health sciences ,Reverse Transcriptase Polymerase Chain Reaction ,Gap junctions, Carotid body, Myenteric plexus, Connexin Cx36, Mouse ,Cell Biology ,General Medicine ,Immunohistochemistry ,Molecular biology ,Mice, Inbred C57BL ,Blot ,Carotid body ,medicine.anatomical_structure ,Real-time polymerase chain reaction ,Cx36 ,Knockout mouse ,sense organs ,030217 neurology & neurosurgery ,Immunostaining - Abstract
Although connexin36 (Cx36) has been studied in several tissues, it is notable that no data are available on Cx36 expression in the carotid body and the intestine. The present study was undertaken to evaluate using immunohistochemistry, PCR and Western blotting procedures, whether Cx36 was expressed in the mouse carotid body and in the intestine at ileum and colon level. In the carotid body, Cx36 was detected as diffuse punctate immunostaining and as protein by Western blotting and mRNA by RT-PCR. Cx36 punctate immunostaining was also evident in the intestine with localization restricted to the myenteric plexus of both the ileum and the colon, and this detection was also confirmed by Western blotting and RT-PCR. All the data obtained were validated using Cx36 knockout mice. Taken together the present data on localization of Cx36 gap-junctions in two tissues of neural crest-derived neuroendocrine organs may provide an anatomical basis for future functional investigations.
- Published
- 2013
149. Identification and functional binding analysis of GPR23/! LPA4 as a candidate G protein-coupled receptor for Guanosine
- Author
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GRILLO, Maria, DI LIBERTO, Valentina, MUDO', Giuseppa, BELLUARDO, Natale, GAROZZO, R, CACIAGLI, F, CONDORELLI, DF, GRILLO, M, DI LIBERTO, V, GAROZZO, R, MUDÒ, G, CACIAGLI, F, CONDORELLI, DF, and BELLUARDO, N
- Subjects
Binding assay ,Guanosine ,Brain ,Gpr23 ,Settore BIO/09 - Fisiologia - Abstract
Several studies have shown that guanine-based purines exert biological effects on the central nervous system (CNS), possibly through membrane receptors, but at the present there are not reports related to the identification of such specific receptor(s). We have identified the first guanosine G protein-coupled receptor GPR23, also known as LPA4 receptor, involved in the modulation of guanosine-mediated antiproliferative effects in human glioma cell line (U87). We report that the silencing of GPR23 reduces significantly the antiproliferative effects of guanosine, while stably transfected cell clones over-expressing GPR23 increase sensitivity to guanosine. [3H] Guanosine radioligand binding assay reveals that [3H]-Guanosine binding to membrane fractions is greatly enhanced by GPR23 overexpression, and inhibited by GPR23 silencing. Furthermore, in [35S] GTPγS binding assay experiments, Guanosine causes a functional G protein coupled receptor activation in U87GPR23 overexp! ressing cells with an EC50= 8,067 nM. The binding site for [3H]-guanosine is highly specific and both lysophosphatidic acid (LPA) and guanine agonists are 10 times less effective than guanosine in displacing 50 nM [3H]-guanosine binding. In order to correlate the effects of guanosine in the CNS to a putative activation of GPR23, we performed, in different brain areas, the following investigations: by PCR, the expression levels of GPR23; by [3H]-Guanosine radioligand binding assay, the binding properties of Guanosine; by [35S] GTPγS binding assay, the receptor activation properties in response to Guanosine. Among the examined brain areas, the cerebral cortex shows the highest GPR23 expression levels which correlate with the highest Bmax values for [3H]-Guanosine as compared to other brain regions, with the following rank order: cerebral cortex>hippocampus>striatum>spinal cord. [35S] GTPγS binding assay experiments confirm an activation of a G protein-cou! pled receptor in response to guanosine in the cerebral cortex (EC50 10 0 nM). Although these observations do not exclude a possible involvement of other unidentified receptors, they can suggest an involvement of GPR23 in the functional response of cerebral cortex to Guanosine. Overall, together these data suggest that GPR23 may act as a functional membrane receptor for Guanosine.
- Published
- 2013
150. The existence of FGFR1-5-HT1A receptor heterocomplexes in midbrain 5-HT neurons of the rat: relevance for neuroplasticity
- Author
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Dasiel O. Borroto-Escuela, Kjell Fuxe, Manuel Narváez, Natale Belluardo, Giuseppa Mudò, Wilber Romero-Fernandez, Francisco Ciruela, Luigi F. Agnati, Alexander O. Tarakanov, Mileidys Pérez-Alea, Universitat de Barcelona, Borroto-Escuela, DO, Romero-Fernandez, W, Perez-Alea, M, Narvaez, M, Tarakanov, AO, Mudò, G, Agnati, LF, Ciruela, F, Belluardo, N, and Fuxe, K
- Subjects
Retracted ,medicine.medical_specialty ,Serotonin receptors ,Encèfal ,Settore BIO/11 - Biologia Molecolare ,Biology ,Settore BIO/09 - Fisiologia ,Receptors de serotonina ,Midbrain ,Internal medicine ,Rates ,medicine ,Receptor ,5-HT receptor ,Neuronal Plasticity, Receptor, Fibroblast Growth Factor, Receptor, Serotonin, 5-HT1A, Serotonergic Neurons, Serotonin ,Raphe ,General Neuroscience ,Encephalon ,Fibroblasts ,Rats ,Endocrinology ,medicine.anatomical_structure ,nervous system ,Forebrain ,Autoreceptor ,5-HT1A receptor ,Neuron - Abstract
The ascending midbrain 5-HT neurons to the forebrain may be dysregulated in depression and have a reduced trophic support. Within situproximity ligation assay (PLA) and supported by coimmunoprecipitation and colocation of the FGFR1 and 5-HT1A immunoreactivities in the midbrain raphe cells, evidence for the existence of FGFR1–5-HT1A receptor heterocomplexes in the dorsal and median raphe nuclei of the Sprague Dawley rat as well as in the rat medullary raphe RN33B cells has been obtained. Especially after combined FGF-2 and 8-OH-DPAT treatment, a marked and significant increase in PLA clusters was found in the RN33B cells. Similar results were reached with the FRET technique in HEK293T cells, where TM-V of the 5HT1A receptor was found to be part of the receptor interface. The combined treatment with FGF-2 and the 5-HT1A agonist also synergistically increased FGFR1 and ERK1/2 phosphorylation in the raphe midline area of the midbrain and the RN33B cells as well as their differentiation, as seen from development of the increased number and length of extensions per cell and their increased 5-HT immunoreactivity. These signaling and differentiation events were dependent on the receptor interface since they were blocked by incubation with TM-V but not by TM-II. Together, the results indicate that the 5-HT1A autoreceptors by being part of a FGFR1–5-HT1A receptor heterocomplex in the midbrain raphe 5-HT nerve cells appear to have a trophic role in the central 5-HT neuron systems in addition to playing a key role in reducing the firing of these neurons.
- Published
- 2012
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