47 results on '"Rostène, W."'
Search Results
2. Down-Regulation of Rat Kidney Calcitonin Receptors by Salmon Calcitonin Infusion Evidenced by Autoradiography
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Bouizar, Z., Rostène, W. H., and Milhaud, G.
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- 1987
3. 3D Distribution of Tyrosine Hydroxylase, Vasopressin and Oxytocin Neurons in the Transparent Postnatal Mouse Brain
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Godefroy, D., Dominici, C., Hardin-Pouzet, H., Anouar, Y., Melik-Parsadaniantz, S., Rostène, W., Goazigo, A. Reaux-Le, Institut de la Vision, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Neuroplasticité des comportements de reproduction = Neuroplasticity of Reproductive Behaviors (NPS-11), Neurosciences Paris Seine (NPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Neuroscience Paris Seine (NPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), and HAL-UPMC, Gestionnaire
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[SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology ,[SDV.NEU.NB] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology - Abstract
International audience; Over the years, advances in immunohistochemistry techniques have been a critical step in detecting and mapping neuromodulatory substances in the central nervous system. The better quality and specificity of primary antibodies, new staining procedures and the spectacular development of imaging technologies have allowed such progress. Very recently, new methods permitting tissue transparency have been successfully used on brain tissues. In this work, we combined whole-mount immunostaining for tyrosine hydroxylase (TH), oxytocin (OXT) and arginine vasopressin (AVP), with iDISCO+ clearing method, light-sheet microscopy and semi automated counting of 3D-labelled neurons to obtain a 3D distribution of these neuronal populations in a 5-day postnatal (P5) mouse brain. Segmentation procedure and 3D reconstruction allowed us to map with high resolution TH staining the various catecholaminergic cell groups and their ascending and descending fiber pathways. We show that TH pathways are present in the whole P5 mouse brain, similar to what was observed in the adult rat brain.
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- 2017
4. CX3CL1 expression in the conjunctiva is involved in immune cell trafficking during toxic ocular surface inflammation
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Denoyer, A, primary, Godefroy, D, additional, Célérier, I, additional, Frugier, J, additional, Riancho, L, additional, Baudouin, F, additional, Rostène, W, additional, and Baudouin, C, additional
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- 2012
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5. CX3CL1 is involved in the ocular surface inflammation induced by benzalkonium chloride
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DENOYER, A, primary, GODEFROY, D, additional, FRUGIER, J, additional, BAUDOUIN, F, additional, ROSTèNE, W, additional, and BAUDOUIN, C, additional
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- 2010
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6. Transcriptional regulation of the tyrosine hydroxylase gene by neurotensin in human neuroblastoma CHP212 cells.
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UCL - MD/FSIO - Département de physiologie et pharmacologie, UCL - MD/GYPE - Département de gynécologie, d'obstétrique et de pédiatrie, UCL - (SLuc) Service de gastro-entérologie et hépatologie pédiatrique, Najimi, Mustapha, Hermans, Emmanuel, Rostène, W, Forgez, P, UCL - MD/FSIO - Département de physiologie et pharmacologie, UCL - MD/GYPE - Département de gynécologie, d'obstétrique et de pédiatrie, UCL - (SLuc) Service de gastro-entérologie et hépatologie pédiatrique, Najimi, Mustapha, Hermans, Emmanuel, Rostène, W, and Forgez, P
- Abstract
The human neuroblastoma cell line CHP212 was found to express functional high affinity neurotensin (NTS-1) receptor subtype. Based on the functional interactions between neurotensin and dopamine transmission, we have used this cell line to investigate the short- and long-term modulation of tyrosine hydroxylase gene expression by the stable neurotensin agonist JMV 449. After exposure of the cells to 1 microM JMV 449 for 5 or 72 h, tyrosine hydroxylase protein and mRNA levels were significantly increased as detected by western blot analysis and quantitative RT-PCR, respectively. Transfection of CHP212 cells with a plasmid containing the luciferase reporter gene under the control of a limited proximal region of the cloned tyrosine hydroxylase promoter, revealed that the effect of JMV 449 results from an increase in the transcriptional activity of the TH gene. These results indicate that modulation of tyrosine hydroxylase gene expression may constitute one of the mechanisms involved in the control of dopamine transmission by neurotensin. Such neurotensin-mediated changes in tyrosine hydroxylase expression may also participate in multiple adaptation processes within the central nervous system to environmental conditions where neurotensin is released such as stress and food intake.
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- 2001
7. Correlative Ultrastructural Distribution of Neurotensin Receptor Proteins and Binding Sites in the Rat Substantia Nigra
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Boudin, H., primary, Pélaprat, D., additional, Rostène, W., additional, Pickel, V. M., additional, and Beaudet, A., additional
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- 1998
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8. Quantitative RT-PCR: Limits and Accuracy
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Souazé, F., primary, Ntodou-Thomé, A., additional, Tran, C.Y., additional, Rostène, W., additional, and Forgez, P., additional
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- 1996
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9. The nonpeptide neurotensin antagonist, SR 48692, used as a tool to reveal putative neurotensin receptor subtypes
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Dubuc, I., primary, Costentin, J., additional, Terranova, J.P., additional, Barnouin, M.C., additional, Soubrié, P., additional, Fur, G. Le, additional, Rostène, W., additional, and Kitabgi, P., additional
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- 1994
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10. Purification, characterization, and cellular localization of the 100-kDa human placental GTPase-activating protein.
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Zhang, Y., primary, Zhang, G., additional, Mollat, P., additional, Carles, C., additional, Riva, M., additional, Frobert, Y., additional, Malassiné, A., additional, Rostène, W., additional, Thang, D.C., additional, and Beltchev, B., additional
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- 1993
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11. Neurotensin agonist induces differential regulation of neurotensin receptor mRNA. Identification of distinct transcriptional and post-transcriptional mechanisms.
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Souazé, F, Rostène, W, and Forgez, P
- Abstract
The binding of neurotensin (NT) to specific receptors triggers the multiple functions that NT exerts in both periphery and brain. By studying the effect of the concentration and time of NT agonist exposure, two separate regulatory mechanisms were detected for the neurotensin receptor (NTR) gene in human colonic adenocarcinoma cells (HT-29). The incubation of cells for 6 h with the NT agonist, JMV 449, resulted in an increase of 270% in NTR mRNA levels. These changes were the direct result of new NTR gene transcription, as indicated by run-on and half-life experiments. In addition, the transcriptional activation of the NTR gene was dependent on NT-receptor complex internalization and de novo protein synthesis. A second response was detected with prolonged exposure to JMV 449. In this case, a decrease of 70% was detected in NTR mRNA levels. Unlike the initial phase, this change was mediated by a post-transcriptional event as the half-life of NTR mRNA from treated cells decreased by 50% as compared with control cells. NT agonist appears to regulate the synthesis of NTR mRNA. In HT-29 cells, this feedback is exerted by a biphasic response. These phases are apparently independent and mediated by two separate mechanisms.
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- 1997
12. Activation of receptor gene transcription is required to maintain cell sensitization after agonist exposure. Study on neurotensin receptor.
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Najimi, M, Souazé, F, Méndez, M, Hermans, E, Berbar, T, Rostène, W, and Forgez, P
- Abstract
Neurotensin (NT) acts through specific G protein-coupled receptors to induce effects in the central nervous system and periphery. In this study we have shown that in the human neuroblastoma cell line CHP 212, an NT agonist, JMV 449, induced high affinity neurotensin receptor (NTR) gene activation. 125I-NT binding of cells challenged with JMV 449 rapidly decreased then reappeared and subsequently stabilized at 50% of the control values after 48 h of agonist exposure. These receptors, which reappeared at the cell surface, are as active as those found in control cells as demonstrated by Ca2+ mobilization. Furthermore, the tyrosine hydroxylase (TH) gene, a known NT target gene, remained activated after prolonged NT agonist exposure in this cell line. In the murine neuroblastoma cell line, N1E-115, NT did not stimulate NTR gene activation but induced NTR mRNA destabilization after long term agonist exposure. In this cell line, NT binding dropped to 15% of control values and remained at this value after agonist treatment. The TH expression, which was originally activated upon NT agonist exposure, decreased to control values after prolonged agonist exposure. These observations combined with the data obtained from a complementary study with HT-29 cells (Souazé, F., Rostène, W., and Forgez, P. (1997) J. Biol. Chem. 272, 10087-10094) revealed the crucial role of agonist-induced receptor gene transcription in the maintenance of cell sensitivity. A model for G protein-coupled receptor regulation induced by prolong and intense agonist stimulation is proposed.
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- 1998
13. Characterization and distribution of binding sites for a new neurotensin receptor antagonist ligand, [3H]SR 48692, in the guinea pig brain
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Catalina Betancur, Canton M, Gully D, Vela G, Pélaprat D, Rostène W, Imagerie cellulaire des neurorécepteurs et physiopathologie neuroendocrinienne, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Sanofi Recherche, SANOFI Recherche, This work was supported by a collaborative grant between INSERM (Institut National de la Santé et de la Recherche Médicale) and Sanofi Recherche. C.B. is a recipient of a Fellowship from the Fondation pour la Recherche Médicale (France), and Betancur, Catalina
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radioligand binding ,Male ,neurotensin receptor ,Guinea Pigs ,neurotensin ,MESH: Binding, Competitive ,Tritium ,MESH: Radioligand Assay ,Binding, Competitive ,Article ,MESH: Guinea Pigs ,SR 48692 ,Iodine Radioisotopes ,MESH: Receptors, Neurotensin ,MESH: Brain ,Radioligand Assay ,receptor autoradiography ,MESH: Autoradiography ,Animals ,Receptors, Neurotensin ,MESH: Animals ,[SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,guinea-pig ,Brain ,MESH: Iodine Radioisotopes ,MESH: Male ,MESH: Tritium ,neurotensin antagonist ,Quinolines ,Autoradiography ,Pyrazoles ,[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,MESH: Quinolines ,MESH: Pyrazoles - Abstract
International audience; SR 48692, a selective nonpeptide antagonist of neurotensin (NT) receptors was developed recently. In the present work we studied the binding properties of the corresponding radioligand, [3H]SR 48692, in the adult guinea pig brain. The characterization of [3H]SR 48692 binding was carried out on brain membrane preparations and the distribution of [3H]SR 48692 binding sites was determined by receptor autoradiography and compared to that of [125I]NT binding sites. In brain homogenates, [3H]SR 48692 bound to a single population of sites with a Kd of 2.19 nM and a maximal binding capacity of 1.15 pmol/mg of protein. This maximal binding capacity value was 20 times higher than that observed for [125I]NT. NT agonists were able to interact competitively with the entire population of binding sites labeled by [3H]SR 48692, but their affinities were much lower than those observed for [125I]NT. By contrast, NT antagonists exhibited similar abilities to inhibit the binding of both radioligands. The addition of unlabeled NT in saturation assays revealed a competitive inhibition of [3H]SR 48692 binding, suggesting that agonist and antagonist ligand bind to overlapping domains of the NT receptor. The autoradiographic distribution of the low-affinity NT binding sites detected by [3H]SR 48692 (96% of the receptors) was very similar to the distribution of high-affinity receptors labeled with [125I]NT (4% of the receptors). In addition, the binding of [3H]SR 48692 was insensitive to guanyl nucleotides. Taken together, these findings suggest that the binding sites detected by [3H]SR 48692 in the guinea pig brain mainly represent the uncoupled form of the NT receptor.
14. Insulin: A 100-Year-Old Discovery With a Fascinating History.
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Rostène W and De Meyts P
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- Animals, Blood Glucose, Dogs, Female, History, 20th Century, Humans, Male, Nobel Prize, Polyuria, Diabetes Mellitus, Insulin
- Abstract
Diabetes has been known since antiquity. We present here a historical perspective on the concepts and ideas regarding the physiopathology of the disease, on the progressive focus on the pancreas, in particular on the islets discovered by Langerhans in 1869, leading to the iconic experiment of Minkowski and von Mering in 1889 showing that pancreatectomy in a dog induced polyuria and diabetes mellitus. Subsequently, multiple investigators searched for the active substance of the pancreas and some managed to produce extracts that lowered blood glucose and decreased polyuria in pancreatectomized dogs but were too toxic to be administered to patients. The breakthrough came 100 years ago, when the team of Frederick Banting, Charles Best, and James Collip working in the Department of Physiology headed by John Macleod at the University of Toronto managed to obtain pancreatic extracts that could be used to treat patients and rescue them from the edge of death by starvation, the only treatment then available. This achievement was quickly recognized by the Nobel Prize in Physiology or Medicine to Banting and Macleod in 1923. At 32, Banting remains the youngest awardee of this prize. Here we discuss the work that led to the discovery and its main breakthroughs, the human characters involved in an increasingly dysfunctional relationship, the controversies that followed the Nobel Prize, and the debate as to who actually "discovered" insulin. We also discuss the early commercial development and progress in insulin crystallization in the decade or so following the Nobel Prize., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2021
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15. Topical treatment with a mu opioid receptor agonist alleviates corneal allodynia and corneal nerve sensitization in mice.
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Joubert F, Guerrero-Moreno A, Fakih D, Reboussin E, Gaveriaux-Ruff C, Acosta MC, Gallar J, Sahel JA, Bodineau L, Baudouin C, Rostène W, Mélik-Parsadaniantz S, and Réaux-Le Goazigo A
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- Administration, Ophthalmic, Analgesics, Opioid administration & dosage, Animals, Cornea drug effects, Cornea innervation, Cornea pathology, Corneal Diseases drug therapy, Corneal Diseases pathology, Disease Models, Animal, Enkephalin, Ala(2)-MePhe(4)-Gly(5)- administration & dosage, Inflammation drug therapy, Inflammation pathology, Male, Mice, Mice, Inbred C57BL, Analgesics, Opioid pharmacology, Enkephalin, Ala(2)-MePhe(4)-Gly(5)- pharmacology, Eye Pain drug therapy, Receptors, Opioid, mu agonists
- Abstract
Corneal pain is considered to be a core symptom of ocular surface disruption and inflammation. The management of this debilitating condition is still a therapeutic challenge. Recent evidence supports a role of the opioid system in the management of corneal nociception. However, the functional involvement of the mu opioid receptor (MOR) underlying this analgesic effect is not known. We first investigated the expression of the MOR in corneal nerve fibers and trigeminal ganglion (TG) neurons in control mice and a mouse model of corneal inflammatory pain. We then evaluated the anti-nociceptive and electrophysiological effects of DAMGO ([D-Ala
2 ,N-Me-Phe4 ,Gly5 -ol] enkephalin), a MOR-selective ligand. MOR immunoreactivity was detected in corneal nerve fibers and primary afferent neurons of the ophthalmic branch of the TG of naive mice. MOR expression was significantly higher in both structures under conditions of inflammatory corneal pain. Topical ocular administration of DAMGO strongly reduced both the mechanical (von Frey) and chemical (capsaicin) corneal hypersensitivity associated with inflammatory ocular pain. Repeated instillations of DAMGO also markedly reversed the elevated spontaneous activity of the ciliary nerve and responsiveness of corneal polymodal nociceptors that were observed in mice with corneal pain. Finally, these DAMGO-induced behavioral and electrophysiological responses were totally blunted by the topical application of naloxone methiodide, an opioid receptor antagonist. Overall, these results provide evidence that topical pharmacological MOR activation may constitute a therapeutic target for the treatment of corneal pain and improve corneal nerve function to alleviate chronic pain., (Copyright © 2020 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.)- Published
- 2020
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16. Chronic dry eye induced corneal hypersensitivity, neuroinflammatory responses, and synaptic plasticity in the mouse trigeminal brainstem.
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Fakih D, Zhao Z, Nicolle P, Reboussin E, Joubert F, Luzu J, Labbé A, Rostène W, Baudouin C, Mélik Parsadaniantz S, and Réaux-Le Goazigo A
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- Animals, Chronic Disease, Female, Inflammation physiopathology, Mice, Mice, Inbred C57BL, Trigeminal Ganglion physiopathology, Cornea physiopathology, Dry Eye Syndromes physiopathology, Hyperalgesia physiopathology, Neuronal Plasticity physiology, Trigeminal Nuclei physiopathology
- Abstract
Background: Dry eye disease (DED) is a multifactorial disease associated with ocular surface inflammation, pain, and nerve abnormalities. We studied the peripheral and central neuroinflammatory responses that occur during persistent DED using molecular, cellular, behavioral, and electrophysiological approaches., Methods: A mouse model of DED was obtained by unilateral excision of the extraorbital lachrymal gland (ELG) and Harderian gland (HG) of adult female C57BL/6 mice. In vivo tests were conducted at 7, 14, and 21 days (d) after surgery. Tear production was measured by a phenol red test and corneal alterations and inflammation were assessed by fluorescein staining and in vivo confocal microscopy. Corneal nerve morphology was evaluated by nerve staining. Mechanical corneal sensitivity was monitored using von Frey filaments. Multi-unit extracellular recording of ciliary nerve fiber activity was used to monitor spontaneous corneal nerve activity. RT-qPCR and immunostaining were used to determine RNA and protein levels at d21., Results: We observed a marked reduction of tear production and the development of corneal inflammation at d7, d14, and d21 post-surgery in DED animals. Chronic DE induced a reduction of intraepithelial corneal nerve terminals. Behavioral and electrophysiological studies showed that the DED animals developed time-dependent mechanical corneal hypersensitivity accompanied by increased spontaneous ciliary nerve fiber electrical activity. Consistent with these findings, DED mice exhibited central presynaptic plasticity, demonstrated by a higher Piccolo immunoreactivity in the ipsilateral trigeminal brainstem sensory complex (TBSC). At d21 post-surgery, mRNA levels of pro-inflammatory (IL-6 and IL-1β), astrocyte (GFAP), and oxidative (iNOS2 and NOX4) markers increased significantly in the ipsilateral trigeminal ganglion (TG). This correlated with an increase in Iba1, GFAP, and ATF3 immunostaining in the ipsilateral TG of DED animals. Furthermore, pro-inflammatory cytokines (IL-6, TNFα, IL-1β, and CCL2), iNOS2, neuronal (ATF3 and FOS), and microglial (CD68 and Itgam) markers were also upregulated in the TBSC of DED animals at d21, along with increased immunoreactivity against GFAP and Iba1., Conclusions: Overall, these data highlight peripheral sensitization and neuroinflammatory responses that participate in the development and maintenance of dry eye-related pain. This model may be useful to identify new analgesic molecules to alleviate ocular pain.
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- 2019
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17. Central CCL2 signaling onto MCH neurons mediates metabolic and behavioral adaptation to inflammation.
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Le Thuc O, Cansell C, Bourourou M, Denis RG, Stobbe K, Devaux N, Guyon A, Cazareth J, Heurteaux C, Rostène W, Luquet S, Blondeau N, Nahon JL, and Rovère C
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- Animals, Chemokine CCL2 deficiency, Chemokine CCL2 immunology, Cytokines biosynthesis, Cytokines genetics, Cytokines immunology, Hypothalamic Hormones genetics, Hypothalamic Hormones immunology, Illness Behavior, Lipopolysaccharides immunology, Melanins genetics, Melanins immunology, Mice, Neurons immunology, Pituitary Hormones genetics, Pituitary Hormones immunology, Receptors, CCR2 metabolism, Weight Loss, Chemokine CCL2 genetics, Chemokine CCL2 metabolism, Hypothalamic Hormones metabolism, Hypothalamus metabolism, Inflammation metabolism, Melanins metabolism, Neurons metabolism, Pituitary Hormones metabolism, Signal Transduction
- Abstract
Sickness behavior defines the endocrine, autonomic, behavioral, and metabolic responses associated with infection. While inflammatory responses were suggested to be instrumental in the loss of appetite and body weight, the molecular underpinning remains unknown. Here, we show that systemic or central lipopolysaccharide (LPS) injection results in specific hypothalamic changes characterized by a precocious increase in the chemokine ligand 2 (CCL2) followed by an increase in pro-inflammatory cytokines and a decrease in the orexigenic neuropeptide melanin-concentrating hormone (MCH). We therefore hypothesized that CCL2 could be the central relay for the loss in body weight induced by the inflammatory signal LPS. We find that central delivery of CCL2 promotes neuroinflammation and the decrease in MCH and body weight. MCH neurons express CCL2 receptor and respond to CCL2 by decreasing both electrical activity and MCH release. Pharmacological or genetic inhibition of CCL2 signaling opposes the response to LPS at both molecular and physiologic levels. We conclude that CCL2 signaling onto MCH neurons represents a core mechanism that relays peripheral inflammation to sickness behavior., (© 2016 The Authors.)
- Published
- 2016
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18. Activation of Prostaglandin FP and EP2 Receptors Differently Modulates Myofibroblast Transition in a Model of Adult Primary Human Trabecular Meshwork Cells.
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Kalouche G, Beguier F, Bakria M, Melik-Parsadaniantz S, Leriche C, Debeir T, Rostène W, Baudouin C, and Vigé X
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- Actins biosynthesis, Actins genetics, Adult, Alprostadil analogs & derivatives, Alprostadil pharmacology, Animals, Antihypertensive Agents pharmacology, Blotting, Western, Cell Survival, Cells, Cultured, Dinoprost, Glaucoma genetics, Glaucoma metabolism, Humans, Immunohistochemistry, Latanoprost, Male, Myofibroblasts drug effects, Myofibroblasts pathology, Myosin Light Chains metabolism, Neuroprotective Agents, Prostaglandins E, Synthetic, Rats, Real-Time Polymerase Chain Reaction, Receptors, Prostaglandin metabolism, Receptors, Prostaglandin E, EP2 Subtype metabolism, Signal Transduction drug effects, Trabecular Meshwork drug effects, Trabecular Meshwork pathology, Glaucoma drug therapy, Myofibroblasts metabolism, Prostaglandins F, Synthetic pharmacology, RNA genetics, Receptors, Prostaglandin drug effects, Receptors, Prostaglandin E, EP2 Subtype drug effects, Trabecular Meshwork metabolism
- Abstract
Purpose: Prostaglandin F2α analogues are the first-line medication for the treatment of ocular hypertension (OHT), and prostanoid EP2 receptor agonists are under clinical development for this indication. The goal of this study was to investigate the effects of F prostanoid (FP) and EP2 receptor activation on the myofibroblast transition of primary trabecular meshwork (TM) cells, which could be a causal mechanism of TM dysfunction in glaucoma., Methods: Human primary TM cells were treated with either latanoprost or butaprost and TGF-β2. Trabecular meshwork contraction was measured in a three-dimensional (3D) TM cell-populated collagen gel (CPCG) model. Expression of α-smooth muscle actin (α-SMA) and phosphorylation of myosin light chain (MLC) were determined by Western blot. Assembly of actin stress fibers and collagen deposition were evaluated by immunocytochemistry. Involvement of p38, extracellular signal-regulated kinase (ERK), and Rho-associated kinase (ROCK) pathways as well as matrix metalloproteinase activation was tested with specific inhibitors., Results: In one source of validated adult TM cells, latanoprost induced cell contraction as observed by CPCG surface reduction and increased actin polymerization, α-SMA expression, and MLC phosphorylation, whereas butaprost inhibited TGF-β2-induced CPCG contraction, actin polymerization, and MLC phosphorylation. Both agonists inhibited TGF-β2-dependent collagen deposition. The latanoprost effects were mediated by p38 pathway., Conclusions: Latanoprost decreased TM collagen accumulation but promoted a contractile phenotype in a source of adult TM cells that could modulate the conventional outflow pathway. In contrast, butaprost attenuated both TM contraction and collagen deposition induced by TGF-β2, thereby inhibiting myofibroblast transition of TM cells. These results open new perspectives for the management of OHT.
- Published
- 2016
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19. Bilateral neuroinflammatory processes in visual pathways induced by unilateral ocular hypertension in the rat.
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Sapienza A, Raveu AL, Reboussin E, Roubeix C, Boucher C, Dégardin J, Godefroy D, Rostène W, Reaux-Le Goazigo A, Baudouin C, and Melik Parsadaniantz S
- Subjects
- Animals, Antigens, CD metabolism, Calcium-Binding Proteins metabolism, Cholera Toxin pharmacokinetics, Cytokines metabolism, Disease Models, Animal, Glial Fibrillary Acidic Protein metabolism, Male, Microfilament Proteins metabolism, Ocular Hypertension pathology, Optometry, Organic Chemicals pharmacokinetics, Proto-Oncogene Proteins c-fos metabolism, Rats, Rats, Long-Evans, Retinal Ganglion Cells pathology, Tumor Necrosis Factor-alpha pharmacology, Up-Regulation drug effects, Visual Pathways metabolism, Encephalitis complications, Encephalitis pathology, Functional Laterality physiology, Ocular Hypertension etiology, Up-Regulation physiology, Visual Pathways pathology
- Abstract
Background: Glaucoma is one of the leading causes of irreversible blindness in the world. The major risk factor is elevated intraocular pressure (IOP) leading to progressive retinal ganglion cell (RGC) death from the optic nerve (ON) to visual pathways in the brain. Glaucoma has been reported to share mechanisms with neurodegenerative disorders. We therefore hypothesize that neuroinflammatory mechanisms in central visual pathways may contribute to the spread of glaucoma disease. The aim of the present study was to analyze the neuroinflammation processes that occur from the pathological retina to the superior colliculi (SCs) in a rat model of unilateral ocular hypertension induced by episcleral vein cauterization (EVC)., Results: Six weeks after unilateral (right eye) EVC in male Long-Evans rats, we evaluated both the neurodegenerative process and the neuroinflammatory state in visual pathway tissues. RGCs immunolabeled (Brn3a(+)) in ipsilateral whole flat-mounted retina demonstrated peripheral RGC loss associated with tissue macrophage/microglia activation (CD68(+)). Gene expression analysis of hypertensive and normotensive retinas revealed a significant increase of pro-inflammatory genes such as CCL2, IL-1β, and Nox2 mRNA expression compared to naïve eyes. Importantly, we found an upregulation of pro-inflammatory markers such as IL-1β and TNFα and astrocyte and tissue macrophage/microglia activation in hypertensive and normotensive RGC projection sites in the SCs compared to a naïve SC. To understand how neuroinflammation in the hypertensive retina is sufficient to damage both right and left SCs and the normotensive retina, we used an inflammatory model consisting in an unilateral stereotaxic injection of TNFα (25 ng/μl) in the right SC of naïve rats. Two weeks after TNFα injection, using an optomotor test, we observed that rats had visual deficiency in both eyes. Furthermore, both SCs showed an upregulation of genes and proteins for astrocytes, microglia, and pro-inflammatory cytokines, notably IL-1β. In addition, both retinas exhibited a significant increase of inflammatory markers compared to a naïve retina., Conclusions: All these data evidence the complex role played by the SCs in the propagation of neuroinflammatory events induced by unilateral ocular hypertension and provide a new insight into the spread of neurodegenerative diseases such as glaucoma.
- Published
- 2016
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20. Opioid and chemokine receptor crosstalk: a promising target for pain therapy?
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Mélik Parsadaniantz S, Rivat C, Rostène W, and Réaux-Le Goazigo A
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- Humans, Pain metabolism, Pain Management, Receptor Cross-Talk physiology, Receptors, Chemokine metabolism, Receptors, Opioid metabolism
- Abstract
Chemokines and opioids are important regulators of immune, inflammatory and neuronal responses in peripheral and central pain pathways. Recent studies have provided insights into the functional interactions between chemokine receptors and opioid receptors, and their role in pain modulation. In this Progress article, we discuss how crosstalk between these two systems might provide a molecular and cellular framework for the development of novel analgesic therapies for the management of acute and/or chronic pain.
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- 2015
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21. Effects of methylmercury contained in a diet mimicking the Wayana Amerindians contamination through fish consumption: mercury accumulation, metallothionein induction, gene expression variations, and role of the chemokine CCL2.
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Bourdineaud JP, Laclau M, Maury-Brachet R, Gonzalez P, Baudrimont M, Mesmer-Dudons N, Fujimura M, Marighetto A, Godefroy D, Rostène W, and Brèthes D
- Subjects
- Adult, Animals, French Guiana, Gene Expression Regulation, Humans, Male, Mice, Mice, Knockout, Organ Specificity, Chemokine CCL2 biosynthesis, Fish Products adverse effects, Food Contamination, Metallothionein biosynthesis, Methylmercury Compounds toxicity
- Abstract
Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutant through fish consumption. We addressed the question of whether a diet mimicking the fish consumption of Wayanas Amerindians from French Guiana could result in observable adverse effects in mice. Wayanas adult men are subjected to a mean mercurial dose of 7 g Hg/week/kg of body weight. We decided to supplement a vegetarian-based mice diet with 0.1% of lyophilized Hoplias aimara fish, which Wayanas are fond of and equivalent to the same dose as that afflicting the Wayanas Amerindians. Total mercury contents were 1.4 ± 0.2 and 5.4 ± 0.5 ng Hg/g of food pellets for the control and aimara diets, respectively. After 14 months of exposure, the body parts and tissues displaying the highest mercury concentration on a dry weight (dw) basis were hair (733 ng/g) and kidney (511 ng/g), followed by the liver (77 ng/g). Surprisingly, despite the fact that MeHg is a neurotoxic compound, the brain accumulated low levels of mercury (35 ng/g in the cortex). The metallothionein (MT) protein concentration only increased in those tissues (kidney, muscles) in which MeHg demethylation had occurred. This can be taken as a molecular sign of divalent mercurial contamination since only Hg(2+) has been reported yet to induce MT accumulation in contaminated tissues. The suppression of the synthesis of the chemokine CCL2 in the corresponding knockout (KO) mice resulted in important changes in gene expression patterns in the liver and brain. After three months of exposure to an aimara-containing diet, eight of 10 genes selected (Sdhb, Cytb, Cox1, Sod1, Sod2, Mt2, Mdr1a and Bax) were repressed in wild-type mice liver whereas none presented a differential expression in KO Ccl2(-/-) mice. In the wild-type mice brain, six of 12 genes selected (Cytb, Cox1, Sod1, Sod2, Mdr1a and Bax) presented a stimulated expression, whereas all remained at the basal level of expression in KO Ccl2(-/-) mice. In the liver of aimara-fed mice, histological alterations were observed for an accumulated mercury concentration as low as 32 ng/g, dw, and metal deposits were observed within the cytoplasm of hepatic cells.
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- 2012
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22. Hyperosmolarity potentiates toxic effects of benzalkonium chloride on conjunctival epithelial cells in vitro.
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Clouzeau C, Godefroy D, Riancho L, Rostène W, Baudouin C, and Brignole-Baudouin F
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- Apoptosis drug effects, Apoptosis Inducing Factor analysis, Caspase 3 analysis, Cell Line, Cell Membrane Permeability drug effects, Cell Survival drug effects, Chromatin metabolism, Conjunctiva pathology, Cytochromes c analysis, Epithelial Cells cytology, Humans, Microscopy, Phase-Contrast, Mitochondria drug effects, Osmolar Concentration, Oxidative Stress, Sodium Chloride chemistry, Xerophthalmia drug therapy, Xerophthalmia pathology, Benzalkonium Compounds adverse effects, Conjunctiva drug effects, Epithelial Cells drug effects, Ophthalmic Solutions adverse effects, Preservatives, Pharmaceutical adverse effects
- Abstract
Purpose: Benzalkonium chloride (BAK), the most commonly used preservative in eye drops, is known to induce ocular irritation symptoms and dry eye in long-term treated patients and animal models. As tear film hyperosmolarity is diagnostic of some types of dry eye disease, we determined in vitro on conjunctival epithelial cells the cytoxicity of BAK in hyperosmolar conditions through cell viability, apoptosis, and oxidative stress assays., Methods: The Wong Kilbourne derivative of Chang conjunctival epithelial cells were cultured for 24 h or 48 h either in NaCl-induced hyperosmolar conditions (400-425-500 mOsM), in low concentrations of BAK (10(-4)%, 3.10(-4)%, and 5.10(-4)%), or in combination of both. We investigated cell viability through lysosomal integrity evaluation, cell death (cell membrane permeability and chromatin condensation), and oxidative stress (reactive oxygen species, superoxide anion) using spectrofluorimetry. Immunohistochemistry was performed for cytoskeleton shrinkage (phalloidin staining), mitochondrial permeability transition pore (cytochrome c release), the apoptosis effector active caspase-3, and the caspase-independent apoptosis factor AIF. We also observed early effects induced by the experimental conditions on the conjunctival cell layers using phase contrast imaging of live cells., Results: As compared to standard culture solutions, hyperosmolar stress potentiated BAK cytotoxicity on conjunctival cells through the induction of oxidative stress; reduction of cell viability; cell membrane permeability increase; cell shrinkage with cell blebbing, as shown in phase contrast imaging of live cells; and chromatin condensation. Like BAK, but to a much lesser extent, hyperosmolarity increased cell death in a concentration-dependent manner through a caspase-dependent apoptosis characterized by a release of cytochrome c in the cytoplasm from mitochondria and the activation of caspase-3. Moreover, the caspase-independent apoptosis factor AIF was found translocated from mitochondria to the nucleus in both conditions., Conclusions: This study showed increased cytotoxic effects of BAK in hyperosmotic conditions, with characteristic cell death processes, namely caspase-dependent and independent apoptosis and oxidative stress. As BAK is known to disrupt tear film, which could promote evaporative dry eye and tear hyperosmolarity, BAK could promote the conditions enhancing its own cytotoxicity. This in vitro hyperosmolarity model thus highlights the risk of inducing a vicious cycle and the importance of avoiding BAK in patients with dry eye conditions.
- Published
- 2012
23. CXCR3 antagonism of SDF-1(5-67) restores trabecular function and prevents retinal neurodegeneration in a rat model of ocular hypertension.
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Denoyer A, Godefroy D, Célérier I, Frugier J, Degardin J, Harrison JK, Brignole-Baudouin F, Picaud S, Baleux F, Sahel JA, Rostène W, and Baudouin C
- Subjects
- Animals, Apoptosis drug effects, Caspase 3 metabolism, Cell Line, Cytoprotection drug effects, Disease Models, Animal, Enzyme Activation drug effects, Glaucoma complications, Glaucoma metabolism, Glaucoma pathology, Glaucoma physiopathology, Humans, Intraocular Pressure drug effects, Male, Rats, Rats, Long-Evans, Receptors, CXCR3 metabolism, Receptors, CXCR4 metabolism, Retinal Degeneration physiopathology, Stress, Physiological drug effects, Trabecular Meshwork drug effects, Trabecular Meshwork pathology, Vision, Ocular drug effects, Chemokine CXCL12 pharmacology, Ocular Hypertension complications, Ocular Hypertension physiopathology, Receptors, CXCR3 antagonists & inhibitors, Retinal Degeneration complications, Retinal Degeneration prevention & control, Trabecular Meshwork physiopathology
- Abstract
Glaucoma, the most common cause of irreversible blindness, is a neuropathy commonly initiated by pathological ocular hypertension due to unknown mechanisms of trabecular meshwork degeneration. Current antiglaucoma therapy does not target the causal trabecular pathology, which may explain why treatment failure is often observed. Here we show that the chemokine CXCL12, its truncated form SDF-1(5-67), and the receptors CXCR4 and CXCR3 are expressed in human glaucomatous trabecular tissue and a human trabecular cell line. SDF-1(5-67) is produced under the control of matrix metallo-proteinases, TNF-α, and TGF-β2, factors known to be involved in glaucoma. CXCL12 protects in vitro trabecular cells from apoptotic death via CXCR4 whereas SDF-1(5-67) induces apoptosis through CXCR3 and caspase activation. Ocular administration of SDF-1(5-67) in the rat increases intraocular pressure. In contrast, administration of a selective CXCR3 antagonist in a rat model of ocular hypertension decreases intraocular pressure, prevents retinal neurodegeneration, and preserves visual function. The protective effect of CXCR3 antagonism is related to restoration of the trabecular function. These data demonstrate that proteolytic cleavage of CXCL12 is involved in trabecular pathophysiology, and that local administration of a selective CXCR3 antagonist may be a beneficial therapeutic strategy for treating ocular hypertension and subsequent retinal degeneration.
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- 2012
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24. Feeding mice with diets containing mercury-contaminated fish flesh from French Guiana: a model for the mercurial intoxication of the Wayana Amerindians.
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Bourdineaud JP, Bellance N, Bénard G, Brèthes D, Fujimura M, Gonzalez P, Marighetto A, Maury-Brachet R, Mormède C, Pédron V, Philippin JN, Rossignol R, Rostène W, Sawada M, and Laclau M
- Subjects
- Adult, Animals, Anxiety chemically induced, Female, French Guiana, Gene Expression, Humans, Indians, South American, Male, Mercury Poisoning genetics, Mercury Poisoning metabolism, Methylmercury Compounds administration & dosage, Methylmercury Compounds pharmacokinetics, Mice, Mice, Inbred C57BL, Middle Aged, Mitochondria, Muscle drug effects, Mitochondria, Muscle metabolism, Mutation, Oxygen Consumption drug effects, Disease Models, Animal, Fishes, Food Contamination, Mercury Poisoning etiology, Methylmercury Compounds poisoning, Methylmercury Compounds toxicity
- Abstract
Background: In 2005, 84% of Wayana Amerindians living in the upper marshes of the Maroni River in French Guiana presented a hair mercury concentration exceeding the limit set up by the World Health Organization (10 microg/g). To determine whether this mercurial contamination was harmful, mice have been fed diets prepared by incorporation of mercury-polluted fish from French Guiana., Methods: Four diets containing 0, 0.1, 1, and 7.5% fish flesh, representing 0, 5, 62, and 520 ng methylmercury per g, respectively, were given to four groups of mice for a month. The lowest fish regimen led to a mercurial contamination pressure of 1 ng mercury per day per g of body weight, which is precisely that affecting the Wayana Amerindians., Results: The expression of several genes was modified with mercury intoxication in liver, kidneys, and hippocampus, even at the lowest tested fish regimen. A net genetic response could be observed for mercury concentrations accumulated within tissues as weak as 0.15 ppm in the liver, 1.4 ppm in the kidneys, and 0.4 ppm in the hippocampus. This last value is in the range of the mercury concentrations found in the brains of chronically exposed patients in the Minamata region or in brains from heavy fish consumers. Mitochondrial respiratory rates showed a 35-40% decrease in respiration for the three contaminated mice groups. In the muscles of mice fed the lightest fish-containing diet, cytochrome c oxidase activity was decreased to 45% of that of the control muscles. When mice behavior was assessed in a cross maze, those fed the lowest and mid-level fish-containing diets developed higher anxiety state behaviors compared to mice fed with control diet., Conclusion: We conclude that a vegetarian diet containing as little as 0.1% of mercury-contaminated fish is able to trigger in mice, after only one month of exposure, disorders presenting all the hallmarks of mercurial contamination.
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- 2008
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25. Cellular and subcellular evidence for neuronal interaction between the chemokine stromal cell-derived factor-1/CXCL 12 and vasopressin: regulation in the hypothalamo-neurohypophysial system of the Brattleboro rats.
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Callewaere C, Fernette B, Raison D, Mechighel P, Burlet A, Calas A, Kitabgi P, Parsadaniantz SM, and Rostène W
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- Animals, Animals, Genetically Modified, Body Water metabolism, Body Water physiology, Chemokine CXCL12 genetics, Chemokine CXCL12 metabolism, Gene Expression Regulation drug effects, Homeostasis genetics, Homeostasis physiology, Hypothalamo-Hypophyseal System drug effects, Hypothalamo-Hypophyseal System metabolism, Hypothalamus chemistry, Hypothalamus metabolism, Male, Pituitary Gland, Posterior metabolism, RNA, Messenger analysis, Rats, Rats, Brattleboro, Rats, Long-Evans, Receptors, CXCR4 genetics, Receptors, CXCR4 metabolism, Subcellular Fractions metabolism, Tissue Distribution, Vasopressins metabolism, Vasopressins pharmacology, Chemokine CXCL12 physiology, Hypothalamo-Hypophyseal System physiology, Neurons metabolism, Neurons physiology, Vasopressins physiology
- Abstract
We previously described a colocalization between arginine vasopressin (AVP) and the chemokine stromal cell-derived factor-1alpha (SDF-1) in the magnocellular neurons of both the hypothalamic supraoptic and paraventricular nucleus as well as the posterior pituitary. SDF-1 physiologically affects the electrophysiological properties of AVP neurons and consequently AVP release. In the present study, we confirm by confocal and electron microscopy that AVP and SDF-1 have a similar cellular distribution inside the neuronal cell and can be found in dense core vesicles in the nerve terminals in the posterior pituitary. Because the Brattleboro rats represent a good model of AVP deficiency, we tested in these animals the fate of SDF-1 and its receptor CXCR4. We identified by immunohistochemistry that both SDF-1 and CXCR4 immunoreactivity were strongly decreased in Brattleboro rats and were strictly correlated with the expression of AVP protein in supraoptic nucleus, paraventricular nucleus, and the posterior pituitary. We observed by real-time PCR an increase in SDF-1 mRNA in both heterozygous and homozygous rats. The effect on the SDF-1/CXCR4 system was not linked to peripheral modifications of kidney water balance because it could not be restored by chronic infusion of deamino-8D-ariginine-vasopressin, an AVP V2-receptor agonist. These original data further suggest that SDF-1 may play an essential role in the regulation of water balance.
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- 2008
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26. Neurotensin receptor antagonist administered during cocaine withdrawal decreases locomotor sensitization and conditioned place preference.
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Felszeghy K, Espinosa JM, Scarna H, Bérod A, Rostène W, and Pélaprat D
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- Analysis of Variance, Animals, Behavior, Animal drug effects, Drug Administration Schedule, Male, Rats, Rats, Sprague-Dawley, Substance Withdrawal Syndrome drug therapy, Time Factors, Cocaine adverse effects, Conditioning, Operant drug effects, Motor Activity drug effects, Pyrazoles administration & dosage, Quinolines administration & dosage, Receptors, Neurotensin antagonists & inhibitors, Substance Withdrawal Syndrome physiopathology
- Abstract
Chronic use of psychostimulants induces enduringly increased responsiveness to a subsequent psychostimulant injection and sensitivity to drug-associated cues, contributing to drug craving and relapse. Neurotensin (NT), a neuropeptide functionally linked to dopaminergic neurons, was suggested to participate in these phenomena. We and others have reported that SR 48692, an NT receptor antagonist, given in pre- or co-treatments with cocaine or amphetamine, alters some behavioral effects of these drugs in rats. However, its efficacy when applied following repeated cocaine administration remains unknown. We, therefore, evaluated the ability of SR 48692, administered after a cocaine regimen, to interfere with the expression of locomotor sensitization and conditioned place preference (CPP) in rats. We demonstrated that the expression of locomotor sensitization, induced by four cocaine injections (15 mg/kg, i.p.) every other day and a cocaine challenge 1 week later, was attenuated by a subsequent 2-week daily administration of SR 48692 (1 mg/kg, i.p.). Furthermore, the expression of cocaine-induced CPP was suppressed by a 10-day SR 48692 treatment started after the conditioning period (four 15 mg/kg cocaine injections every other day). Taken together, our data show that a chronic SR 48692 treatment given after a cocaine regimen partly reverses the expression of locomotor sensitization and CPP in the rat, suggesting that NT participates in the maintenance of these behaviors. Our results support the hypothesis that targeting neuromodulatory systems, such as the NT systems may offer new strategies in the treatment of drug addiction.
- Published
- 2007
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27. Chemokines: a new class of neuromodulator?
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Rostène W, Kitabgi P, and Parsadaniantz SM
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- Animals, Brain embryology, Humans, Neurosecretory Systems physiology, Neurotransmitter Agents physiology, Brain physiology, Chemokines physiology, Receptors, Chemokine physiology
- Abstract
Chemokines are not only found in the immune system or expressed in inflammatory conditions: they are constitutively present in the brain in both glial cells and neurons. Recently, the possibility has been raised that they might act as neurotransmitters or neuromodulators. Although the evidence is incomplete, emerging data show that chemokines have several of the characteristics that define neurotransmitters. Moreover, their physiological actions resemble those of neuromodulators in the sense that chemokines usually have few effects by themselves in basal conditions, but modify the induced release of neurotransmitters or neuropeptides. These findings, together with the pharmacological development of agonists and antagonists that are selective for chemokine receptors and can cross the blood-brain barrier, open a new era of research in neuroscience.
- Published
- 2007
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28. Chemokines and chemokine receptors in the brain: implication in neuroendocrine regulation.
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Callewaere C, Banisadr G, Rostène W, and Parsadaniantz SM
- Subjects
- Chemokines chemistry, Humans, Protein Conformation, Chemokines physiology, Neurosecretory Systems physiology, Receptors, Chemokine physiology
- Abstract
Chemokines are small secreted proteins that chemoattract and activate immune and non-immune cells both in vivo and in vitro. In addition to their well-established role in the immune system, several recent reports have suggested that chemokines and their receptors may also play a role in the central nervous system (CNS). The best known central action is their ability to act as immunoinflammatory mediators. Indeed, these proteins regulate leukocyte infiltration in the brain during inflammatory and infectious diseases. However, we and others recently demonstrated that they are expressed not only in neuroinflammatory conditions, but also constitutively by different cell types including neurons in the normal brain, suggesting that they may act as modulators of neuronal functions. The goal of this review is to highlight the role of chemokines in the control of neuroendocrine functions. First, we will focus on the expression of chemokines and their receptors in the CNS, with the main spotlight on the neuronal expression in the hypothalamo-pituitary system. Secondly, we will discuss the role--we can now suspect--of chemokines and their receptors in the regulation of neuroendocrine functions. In conclusion, we propose that chemokines can be added to the well-described neuroendocrine regulatory mechanisms, providing an additional fine modulatory tuning system in physiological conditions.
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- 2007
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29. Neurotensin receptor 1 gene activation by the Tcf/beta-catenin pathway is an early event in human colonic adenomas.
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Souazé F, Viardot-Foucault V, Roullet N, Toy-Miou-Leong M, Gompel A, Bruyneel E, Comperat E, Faux MC, Mareel M, Rostène W, Fléjou JF, Gespach C, and Forgez P
- Subjects
- Adenoma physiopathology, Adenomatous Polyposis Coli Protein physiology, Cell Proliferation, Cell Survival, Cell Transformation, Neoplastic, Colonic Neoplasms physiopathology, Humans, Loss of Heterozygosity, Promoter Regions, Genetic, Receptors, Neurotensin physiology, Signal Transduction, Up-Regulation, Wnt Proteins physiology, Adenoma genetics, Colonic Neoplasms genetics, Receptors, Neurotensin biosynthesis, TCF Transcription Factors physiology, beta Catenin physiology
- Abstract
Alterations in the Wnt/APC (adenomatous polyposis coli) signalling pathway, resulting in beta-catenin/T cell factor (Tcf)-dependent transcriptional gene activation, are frequently detected in familial and sporadic colon cancers. The neuropeptide neurotensin (NT) is widely distributed in the gastrointestinal tract. Its proliferative and survival effects are mediated by a G-protein coupled receptor, the NT1 receptor. NT1 receptor is not expressed in normal colon epithelial cells, but is over expressed in a number of cancer cells and tissues suggesting a link to the outgrowth of human colon cancer. Our results demonstrate that the upregulation of NT1 receptor occurring in colon cancer is the result of Wnt/APC signalling pathway activation. We first established the functionality of the Tcf response element within the NT1 receptor promoter. Consequently, we observed the activation of NT1 receptor gene by agents causing beta-catenin cytosolic accumulation, as well as a strong decline of endogenous receptor when wt-APC was restored. At the cellular level, the re-establishment of wt-APC phenotype resulted in the impaired functionality of NT1 receptor, like the breakdown in NT-induced intracellular calcium mobilization and the loss of NT pro-invasive effect. We corroborated the Wnt/APC signalling pathway on the NT1 receptor promoter activation with human colon carcinogenesis, and showed that NT1 receptor gene activation was perfectly correlated with nuclear or cytoplasmic beta-catenin localization while NT1 receptor was absent when beta-catenin was localized at the cell-cell junction in early adenomas of patients with familial adenomatous polyposis, hereditary non-polyposis colorectal cancer and loss of heterozygosity tumours. In this report we establish a novel link in vitro between the Tcf/beta-catenin pathway and NT1 receptor promoter activation.
- Published
- 2006
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30. [Claude Fortier: the great history of neuroendocrinology].
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Rostène W
- Subjects
- History, 20th Century, Quebec, Stress, Psychological, Neuroendocrinology history
- Abstract
The work of Claude Fortier is linked to the history of neuroendocrinology. Through him and his pioneer work in Montreal with Hans Selye, the << Man of stress >>, and at Laval University in Quebec City in his own laboratory, where all researchers involved in the study of the hypothalamo-hypophysial adrenal axis have been through, it is the whole saga of the search for the neuropeptide CRH (corticotropin releasing factor), and the harsh fight for the Nobel distinction that can be related. Among Claude Fortier's scientific discoveries, the feedback mechanisms of glucocorticoid hormones on brain and pituitary function, the presence of both mineralo and glucocorticoid receptors in some brain structures, and the introduction of computer science in biomedical research, can be cited. The consequences of these discoveries are illustrated in the pathologies linked to stress (anxiety, depression, addiction). Claude Fortier was not only a great figure in biomedical science, honored by several distinctions, but also an important personality in the policy of research in which he played a prominent role in Quebec medical research and allowed it to rank among the best in the world.
- Published
- 2005
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31. Death-associated protein kinase loss of expression is a new marker for breast cancer prognosis.
- Author
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Lévy D, Plu-Bureau G, Decroix Y, Hugol D, Rostène W, Kimchi A, and Gompel A
- Subjects
- Antineoplastic Agents, Hormonal therapeutic use, Apoptosis drug effects, Apoptosis Regulatory Proteins, Biomarkers, Tumor biosynthesis, Breast cytology, Breast drug effects, Breast enzymology, Breast Neoplasms drug therapy, Breast Neoplasms enzymology, Cell Division drug effects, Cell Line, Tumor, Cells, Cultured, Death-Associated Protein Kinases, Epithelial Cells drug effects, Epithelial Cells enzymology, Estradiol analogs & derivatives, Estradiol pharmacology, Estrogen Receptor Modulators pharmacology, Female, Humans, Immunohistochemistry, Prognosis, Proportional Hazards Models, Proto-Oncogene Proteins c-bcl-2 analysis, Receptors, Estrogen analysis, Receptors, Progesterone analysis, Survival Analysis, Tamoxifen therapeutic use, Breast Neoplasms pathology, Calcium-Calmodulin-Dependent Protein Kinases biosynthesis
- Abstract
Purpose: Death-associated protein (DAP)-kinase is a new Ser/Thr kinase involved in cell apoptosis and tumor suppression, the expression of which has been correlated to invasive potential and metastasis in several human neoplastic tissues. We analyzed the level of DAP-kinase expression in breast cancer specimens and its correlation with survival., Experimental Design: One hundred twenty-eight breast cancer specimens were analyzed by immunohistochemistry. Patient records were studied retrospectively for demographic characteristics, clinical data, hormonal treatment, outcome, and survival. DAP-kinase protein expression was also studied in normal breast cells primary cultures under estrogen and antiestrogen treatment., Results: Among the 128 patients, 30 showed a DAP-kinase staining < or = 20%, whereas 98 had a staining over 20%. Mean follow-up time was 62 months. The association between tumor Scarff-Bloom and Richardson grade (P = 0.009), estrogen receptor and progesterone receptor expression (P = 0.002 and 0.001, respectively), tumor size (P = 0.05), Bcl-2 expression (P = 0.004), and DAP-kinase immunostaining in the ductal carcinoma group was highly significant. Overall (64 months) and disease-free (63 months) survival in the high DAP-kinase expression group were significantly longer compared with the women whose tumors showed a loss of DAP-kinase expression (51 and 43 months, respectively). DAP-kinase protein was strongly expressed in normal breast tissue and in human breast epithelial cells primary cultures. Estradiol decreased DAP-kinase expression in these cells, arguing for hormonal regulation of the protein., Conclusions: Loss of DAP-kinase expression negatively correlates to survival and positively correlates to the probability of recurrence in a very significant manner. DAP-kinase thus constitutes a novel and independent prognosis marker for breast cancer.
- Published
- 2004
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32. Receptor trafficking via the perinuclear recycling compartment accompanied by cell division is necessary for permanent neurotensin cell sensitization and leads to chronic mitogen-activated protein kinase activation.
- Author
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Toy-Miou-Leong M, Cortes CL, Beaudet A, Rostène W, and Forgez P
- Subjects
- Animals, Biological Transport, CHO Cells, Cell Differentiation, Cell Line, Tumor, Cell Membrane metabolism, Cell Nucleus metabolism, Cricetinae, Cytoplasm metabolism, Disease Progression, Enzyme Activation, Gene Expression Regulation, Neoplastic, Green Fluorescent Proteins, Humans, Immunohistochemistry, Ligands, Luminescent Proteins metabolism, Lysosomes metabolism, Mice, Microscopy, Confocal, Microscopy, Fluorescence, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase 3, Mitogen-Activated Protein Kinases metabolism, Neurotensin agonists, Phosphorylation, Protein Binding, Time Factors, Transfection, Transferrin metabolism, Xanthenes pharmacology, MAP Kinase Signaling System, Neurotensin chemistry, Neurotensin physiology, Receptors, Neurotensin chemistry
- Abstract
Most G protein-coupled receptors are internalized after interaction with their respective ligand, a process that subsequently contributes to cell desensitization, receptor endocytosis, trafficking, and finally cell resensitization. Although cellular mechanisms leading to cell desensitization have been widely studied, those responsible for cell resensitization are still poorly understood. We examined here the traffic of the high affinity neurotensin receptor (NT1 receptor) following prolonged exposure to high agonist concentration. Fluorescence and confocal microscopy of Chinese hamster ovary, human neuroblastoma (CHP 212), and murine neuroblastoma (N1E-115) cells expressing green fluorescent protein-tagged NT1 receptor revealed that under prolonged treatment with saturating concentrations of neurotensin (NT) agonist, NT1 receptor and NT transiently accumulated in the perinuclear recycling compartment (PNRC). During this cellular event, cell surface receptors remained markedly depleted as detected by both confocal microscopy and (125)I-NT binding assays. In dividing cells, we observed that following prolonged NT agonist stimulation, NT1 receptors were removed from the PNRC, accumulated in dispersed vesicles inside the cytoplasm, and subsequently reappeared at the cell surface. This NT binding recovery allowed for constant cell sensitization and led to a chronic activation of mitogen-activated protein kinases p42 and p44. Under these conditions, the constant activation of NT1 receptor generates an oncogenic regulation. These observations support the potent role for neuropeptides, such as NT, in cancer progression.
- Published
- 2004
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33. NT agonist regulates expression of nuclear high-affinity neurotensin receptors.
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Toy-Miou-Leong M, Bachelet CM, Pélaprat D, Rostène W, and Forgez P
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- Animals, Binding Sites, CHO Cells, Cell Line, Tumor, Cell Nucleus metabolism, Cricetinae, Fluorescent Antibody Technique, Gene Expression Regulation, Humans, In Vitro Techniques, Lung cytology, Lung metabolism, Oligopeptides pharmacology, Radioligand Assay, Rats, Rats, Sprague-Dawley, Receptors, Neurotensin agonists, Receptors, Neurotensin genetics, Reverse Transcriptase Polymerase Chain Reaction, Substantia Nigra metabolism, Substantia Nigra ultrastructure, Cell Nucleus drug effects, Receptors, Neurotensin biosynthesis
- Abstract
Neurotensin (NT) exerts multiple functions in the central nervous system and peripheral tissues. Its actions are mainly mediated by a high-affinity G-protein-coupled receptor, the NT-1 receptor. In this study we demonstrated a nuclear NT binding site in different cellular models. We first noted that a large percentage of NT-1 receptor cell body immunoreactivity was located in the nuclear soma and nuclear envelope of rat substantia nigra, a brain area rich in NT-containing axon terminals. The NT-1 receptor was also visualized in purified nuclei from CHO cells stably transfected with NT-1 receptor coupled to the enhanced green fluorescence protein by immunocytochemistry. We observed that both the nuclear envelope and the nuclear soma were labeled, and the labeling intensity significantly increased after NT agonist treatment. These results suggested that NT-1 receptors, present in both the nuclear soma and the nuclear envelope, can be modulated by the ligand. Lastly, [(125)I]-NT binding experiments performed on isolated nuclei from a human lung cancer cell line endogenously expressing NT-1 receptor and NT, LNM35, revealed the existence of nuclear Gpp(NHp)-sensitive binding sites. These binding sites markedly decreased when cells were chronically treated with an NT-1 receptor antagonist, SR 48692. Taken together, these data suggest that the agonist regulates the expression of nuclear NT-1 receptors.
- Published
- 2004
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34. Characterization and functionality of CXCR4 chemokine receptor and SDF-1 in human corneal fibroblasts.
- Author
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Bourcier T, Berbar T, Paquet S, Rondeau N, Thomas F, Borderie V, Laroche L, Rostène W, Haour F, and Lombet A
- Subjects
- Autoradiography, Binding Sites, Blotting, Western, Calcium metabolism, Cell Membrane metabolism, Cells, Cultured, Chemokine CXCL12, Chemokines, CXC genetics, Cornea cytology, Fibroblasts metabolism, Fura-2 metabolism, Humans, Immunoblotting, RNA, Messenger metabolism, Receptors, CXCR4 genetics, Reverse Transcriptase Polymerase Chain Reaction, Stromal Cells metabolism, Chemokines, CXC metabolism, Cornea metabolism, Fura-2 analogs & derivatives, Receptors, CXCR4 metabolism
- Abstract
Purpose: The aim of this study was to investigate whether cultured human corneal fibroblasts express functional chemokine CXCR4 receptors on their cell surface and to determine the presence of its specific ligand, SDF-1 (CXCL12), in human corneal fibroblasts., Methods: Human corneal fibroblast cultures were obtained using human donor corneas. CXCR4 receptors were characterized using binding studies and autoradiography with [125I]SDF-1. The functionality of CXCR4 receptors was assessed by intracellular calcium measurement using a dynamic imaging microscopy system. CXCR4 and SDF-1 mRNA were detected in human corneal fibroblasts using reverse transcriptase polymerase chain reaction (RT-PCR). The CXCR4 protein was detected by western blot analysis., Results: [125I]SDF-1 specifically bound to cultured corneal fibroblasts with a KD value of 8.3+/-1.2 nM. The presence of CXCR4 was confirmed by autoradiography of the radioligand on slices of corneal stroma. SDF-1 induced a rapid and transient intracellular calcium increase in cultured corneal fibroblasts that was blocked by the specific antagonist bicyclam. Moreover, a 48 kDa protein was detected by western blot analysis of corneal fibroblast extracts, using a specific CXCR4 polyclonal antibody. RT-PCR showed the expression of both CXCR4 and SDF-1 mRNAs in human corneal fibroblasts., Conclusions: These results indicate for the first time that cultured human corneal fibroblasts express the chemokine receptors CXCR4 and its ligand SDF-1. This latter might exert physiological effects on the cornea and could be involved in pathological conditions such as corneal angiogenesis.
- Published
- 2003
35. Expression of neurotensin receptors in human corneal keratocytes.
- Author
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Bourcier T, Rondeau N, Paquet S, Forgez P, Lombet A, Pouzaud F, Rostène W, Borderie V, and Laroche L
- Subjects
- Apoptosis, Blotting, Western, Calcium metabolism, Cell Division, Cell Survival, Cells, Cultured, DNA Primers chemistry, Humans, RNA, Messenger metabolism, Reverse Transcriptase Polymerase Chain Reaction, Corneal Stroma cytology, Fibroblasts metabolism, Receptors, Neurotensin genetics, Receptors, Neurotensin metabolism
- Abstract
Purpose: The purpose of the study was to investigate whether cultured human keratocytes express the neurotensin receptors (NTR1, NTR2, and NTR3), to determine the presence of neurotensin (NT) in keratocytes, and to assess the influence of NT on these cells., Methods: Human keratocytes were cultured in medium treated with various concentrations (10(-7)-10(-9) M) of JMV449 (a weakly degradable NT agonist). Cell proliferation and viability were analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay. Apoptosis was studied by nucleus labeling with a fluorescent dye and cold light fluorometry. NT, NTR1, NTR2, and NTR3 mRNA were detected in human keratocytes by means of reverse transcriptase-polymerase chain reaction (RT-PCR). NTR1 protein was detected by Western blot analysis. Functionality of NTR1 was assessed by intracellular calcium ([Ca2+]i) measurement with a dynamic imaging microscopy system., Results: RT-PCR and Western blot analysis showed the expression of the NTR1 (mRNA and protein) and NTR3 mRNA in human corneal keratocytes. NT and NTR2 mRNA were undetectable. JMV449 induced a rapid and transient [Ca2+]i increase in human corneal keratocytes that was blocked by the specific antagonist SR48692. JMV449 significantly increased cell proliferation and viability after 72, 96, and 120 hours of culture, with a maximum effect at 10(-7) M (P < 0.005). Finally, JMV449 decreased keratocyte apoptosis, whatever the concentration used (P < 0.005)., Conclusions: These results indicate that cultured human keratocytes express NTR1 and NTR3 and that NT may exert physiological effects on cornea such as regulation of keratocyte proliferation and apoptosis.
- Published
- 2002
36. Resistance to induced apoptosis in the human neuroblastoma cell line SK-N-SH in relation to neuronal differentiation. Role of Bcl-2 protein family.
- Author
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Lombet A, Zujovic V, Kandouz M, Billardon C, Carvajal-Gonzalez S, Gompel A, and Rostène W
- Subjects
- Androstadienes pharmacology, Bacterial Toxins pharmacology, Biomarkers analysis, Cell Size drug effects, DNA Fragmentation drug effects, Gene Expression Regulation drug effects, Humans, Microtubule-Associated Proteins metabolism, Neuroblastoma metabolism, Neurons drug effects, Neurons metabolism, Phosphatidylinositol 3-Kinases metabolism, Phosphoinositide-3 Kinase Inhibitors, Phosphopyruvate Hydratase metabolism, Protein Kinase C antagonists & inhibitors, Protein Kinase C metabolism, Signal Transduction drug effects, Staurosporine pharmacology, Synaptophysin metabolism, Tetradecanoylphorbol Acetate pharmacology, Thapsigargin pharmacology, Tretinoin pharmacology, Tumor Cells, Cultured, Wortmannin, bcl-X Protein, rhoA GTP-Binding Protein metabolism, Apoptosis drug effects, Bacterial Proteins, Cell Differentiation drug effects, Neuroblastoma pathology, Neurons pathology, Proto-Oncogene Proteins c-bcl-2 metabolism
- Abstract
Much evidence suggests that apoptosis plays a crucial role in cell population homeostasis that depends on the expression of various genes implicated in the control of cell life and death. The sensitivity of human neuroblastoma cells SK-N-SH to undergo apoptosis induced by thapsigargin was examined. SK-N-SH were previously differentiated into neuronal cells by treatments with retinoic acid (RA), 4 beta-phorbol 12-myristate 13-acetate (PMA) which increases protein kinase C (PKC) activity, and staurosporine which decreases PKC activity. Neuronal differentiation was evaluated by gamma-enolase, microtubule associated protein 2 (MAP2) and synaptophysin immunocytochemistry. The sensitivity of the cells to thapsigargin-induced apoptosis was evaluated by cell viability and nuclear fragmentation (Hoechst 33258) and compared with pro-(Bcl-2, Bcl-x(L)) and anti-apoptotic (Bax, Bak) protein expression of the Bcl-2 family. Cells treated with RA and PMA were more resistant to apoptosis than controls. Conversely, the cells treated with staurosporine were more susceptible to apoptosis. In parallel with morphological modifications, the expression of inhibitors and activators of apoptosis was directly dependent upon the differentiating agent used. Bcl-2 expression was strongly increased by PMA and drastically decreased by staurosporine as was Bcl-x(L) expression. Bax and Bak expression were not significantly modified. These results demonstrate that drugs that modulate PKC activity may induce a modification of Bcl-2 expression as well as resistance to the apoptotic process. Furthermore, the expression of Bcl-2 was reduced by toxin B from Clostridium difficile and, to a lesser extent, by wortmannin suggesting a role of small G-protein RhoA and PtdIns3 kinase in the control of Bcl-2 expression. Our data demonstrate a relationship between the continuous activation of PKC, the expression of Bcl-2 protein family and the resistance of differentiated SK-N-SH to apoptosis.
- Published
- 2001
- Full Text
- View/download PDF
37. Neurotensin gene expression and behavioral responses following administration of psychostimulants and antipsychotic drugs in dopamine D(3) receptor deficient mice.
- Author
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Betancur C, Lépée-Lorgeoux I, Cazillis M, Accili D, Fuchs S, and Rostène W
- Subjects
- Amphetamine pharmacology, Animals, Brain metabolism, Catalepsy chemically induced, Cocaine pharmacology, Female, Gene Expression physiology, Haloperidol pharmacology, Male, Mice, Mice, Mutant Strains, Motor Activity physiology, Neurotensin genetics, Neurotensin metabolism, RNA, Messenger drug effects, RNA, Messenger metabolism, Receptors, Dopamine D2 deficiency, Receptors, Dopamine D2 metabolism, Receptors, Dopamine D3, Antipsychotic Agents pharmacology, Brain drug effects, Dopamine Uptake Inhibitors pharmacology, Gene Expression drug effects, Motor Activity drug effects, Neurotensin drug effects, Receptors, Dopamine D2 drug effects
- Abstract
Exposure to psychostimulants and antipsychotics increases neurotensin (NT) gene expression in the striatum and nucleus accumbens. To investigate the contribution of D(3) receptors to these effects we used mice with targeted disruption of the D(3) receptor gene. Basal NT mRNA expression was similar in D(3) receptor mutant mice and wild-type animals. Acute administration of haloperidol increased NT gene expression in the striatum in D(3)+/+, D(3)+/- and D(3)-/- mice. Similarly, acute cocaine and amphetamine induced NT mRNA expression in the nucleus accumbens shell and olfactory tubercle to a comparable extent in D(3) mutants and wild-type mice. Daily injection of cocaine for seven days increased NT mRNA in a restricted population of neurons in the dorsomedial caudal striatum of D(3)+/+ mice, but not in D(3)-/- and D(3)+/- animals. No differences were observed between D(3) receptor mutant mice and wild-type littermates in the locomotor activity and stereotyped behaviors induced by repeated cocaine administration. These findings demonstrate that dopamine D(3) receptors are not necessary for the acute NT mRNA response to drugs of abuse and antipsychotics but appear to play a role in the regulation of NT gene induction in striatal neurons after repeated cocaine. In addition, our results indicate that the acute locomotor response to cocaine and development of psychostimulant-induced behavioral sensitization do not require functional D(3) receptors.
- Published
- 2001
- Full Text
- View/download PDF
38. Regulation of human corneal epithelial cell proliferation and apoptosis by dexamethasone.
- Author
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Bourcier T, Forgez P, Borderie V, Scheer S, Rostène W, and Laroche L
- Subjects
- Cells, Cultured, DNA Primers chemistry, Epithelium, Corneal drug effects, Gene Expression, Humans, Immunoenzyme Techniques, Methylphenazonium Methosulfate metabolism, RNA, Messenger metabolism, Receptors, Glucocorticoid biosynthesis, Reverse Transcriptase Polymerase Chain Reaction, Tetrazolium Salts metabolism, Thiazoles metabolism, Apoptosis drug effects, Cell Division drug effects, Dexamethasone pharmacology, Epithelium, Corneal cytology, Epithelium, Corneal metabolism, Glucocorticoids pharmacology, Receptors, Glucocorticoid genetics
- Abstract
Purpose: To investigate whether human corneal epithelial cells express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells., Methods: Human corneal epithelial cells were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfop henyl) -2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Apoptosis was studied by nucleus labeling using a fluorescent dye and immunostaining by APO 2.7 at 6 days of culture. GR mRNA was detected in corneal epithelium and cultured corneal epithelial cells by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the epithelial cells was performed with a monoclonal anti-human GR., Results: RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in corneal epithelial cells. DEX significantly increased corneal epithelial cell proliferation with concentrations ranging from 10(-10) to 10(-6) M, with a maximum effect at 10(-7) M (P < 0.005). However, DEX also induced apoptosis of cultured corneal epithelial cells at any concentration used., Conclusions: These results indicate that human corneal epithelial cells express the GR and proliferate in response to DEX stimulation which also induces corneal epithelial cell apoptosis.
- Published
- 2000
39. Successful surgical removal of occult metastases of medullary thyroid carcinoma recurrences with the help of immunoscintigraphy and radioimmunoguided surgery.
- Author
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de Labriolle-Vaylet C, Cattan P, Sarfati E, Wioland M, Billotey C, Brochériou C, Rouvier E, de Roquancourt A, Rostène W, Askienazy S, Barbet J, Milhaud G, and Gruaz-Guyon A
- Subjects
- Adult, Aged, Calcitonin analysis, Carcinoembryonic Antigen blood, Carcinoma, Medullary diagnostic imaging, Carcinoma, Medullary pathology, Carcinoma, Medullary surgery, False Negative Reactions, False Positive Reactions, Female, Humans, Lymphatic Metastasis, Male, Middle Aged, Neoplasm Metastasis, Recurrence, Reproducibility of Results, Sensitivity and Specificity, Thyroid Neoplasms pathology, Thyroid Neoplasms surgery, Carcinoma, Medullary secondary, Radioimmunodetection, Thyroid Neoplasms diagnostic imaging
- Abstract
Patients with recurrent or metastatic medullary thyroid carcinoma (MTC) were referred for pretargeted immunoscintigraphy (Affinity Enhancement System; AES) and radioimmunoguided surgery (RIGS). Data collected from 13 patients establish that whole-body AES immunoscintigraphy revealed metastases < 360 mg and RIGS detected micrometastases (5-15 mg). All tissue samples removed by the surgeon were diagnosed by histology and immunohistochemistry of calcitonin to check the accuracy of IS and RIGS results. AES immunoscintigraphy is very sensitive. Of 34 metastases or recurrences detected, 22 had escaped physical examination or conventional imaging. The accuracy of RIGS was 86%, its sensitivity 75%, and its specificity was 90% (n = 208). IS and RIGS detected occult tumors that would have escaped surgery, clearly demonstrating clinical benefit. Serum calcitonin (normal, 10 pg/ml) and carcinoembryonic antigen (normal, 5 ng/ml) of two patients were restored to normal. In patients whose tumors were discovered, progression of their disease was slowed, as evidenced by the large decrease in serum calcitonin and carcinoembryonic antigen, an important prognostic factor. Surgery was canceled in one case where IS detected distant metastases out of surgical reach. Thus, AES immunoscintigraphy and RIGS might be of valuable help for the surgical management of medullary thyroid carcinoma.
- Published
- 2000
40. In vitro effects of dexamethasone on human corneal keratocytes.
- Author
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Bourcier T, Borderie V, Forgez P, Lombet A, Rostène W, and Laroche L
- Subjects
- Apoptosis drug effects, Cell Division drug effects, Cells, Cultured, Cornea cytology, Cornea metabolism, Dexamethasone antagonists & inhibitors, Glucocorticoids antagonists & inhibitors, Hormone Antagonists pharmacology, Humans, Immunohistochemistry, Mifepristone pharmacology, Osmolar Concentration, Receptors, Glucocorticoid genetics, Receptors, Glucocorticoid metabolism, Reverse Transcriptase Polymerase Chain Reaction, Cornea drug effects, Dexamethasone pharmacology, Glucocorticoids pharmacology
- Abstract
Purpose: To investigate whether cultured human keratocytes express the glucocorticoid receptor (GR) and to assess the influence of dexamethasone (DEX) on these cells., Methods: Human keratocytes were cultured in medium supplemented with various concentrations of DEX (ranging from 10(-10) to 10(-4) M). Cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-s ulfophenyl)-2H-tetrazolium inner salt (MTS) assay at 2, 4, and 6 days of culture. Some experiments were performed in the presence of mifepristone (RU38486), an antiglucocorticoid molecule. The early phase of apoptosis was studied by means of keratocyte staining with a fluorescein conjugate of annexin V and propidium iodide, and cells were analyzed by flow cytometry. Glucocorticoid receptor mRNA was detected in keratocytes by means of reverse transcription-polymerase chain reaction (RT-PCR). Immunocytochemical staining of the cells was performed with a monoclonal anti-human GR., Results: RT-PCR and immunocytochemistry showed the expression of GR (mRNA and protein) in cultured keratocytes. Dexamethasone significantly increased keratocyte proliferation with concentrations ranging from 10(-9) to 10(-5) M, with a maximum effect at 10(-7) M (P < 0.005). Dexamethasone's proproliferative effect was inhibited by RU38486. However, DEX also induced apoptosis of cultured keratocytes at any concentration used., Conclusions: These results indicate that cultured human keratocytes express the GR and proliferate in response to DEX stimulation (10(-9)-10(-5) M), which also induces keratocyte apoptosis.
- Published
- 1999
41. Role of endogenous neurotensin in the behavioral and neuroendocrine effects of cocaine.
- Author
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Betancur C, Cabrera R, de Kloet ER, Pélaprat D, and Rostène W
- Subjects
- Animals, Hypothalamo-Hypophyseal System physiology, Male, Motor Activity physiology, Pituitary-Adrenal System physiology, Pyrazoles pharmacology, Quinolines pharmacology, Rats, Rats, Sprague-Dawley, Receptors, Neurotensin physiology, Stereotyped Behavior physiology, Cocaine pharmacology, Corticosterone blood, Dopamine Uptake Inhibitors pharmacology, Hypothalamo-Hypophyseal System drug effects, Motor Activity drug effects, Neurotensin physiology, Pituitary-Adrenal System drug effects, Receptors, Neurotensin antagonists & inhibitors, Stereotyped Behavior drug effects
- Abstract
The present experiments were designed to assess the role of endogenous neurotensin (NT) in the behavioral response to acute and daily cocaine, after administration of the NT receptor antagonist, SR 48692. Given that glucocorticoids increase the sensitivity to the psychomotor effects of drugs of abuse, we also investigated the effects of SR 48692 on basal and cocaine-induced corticosterone secretion. Acute administration of SR 48692 (1 mg/kg i.p.) reduced the number of rearings induced by cocaine (15 mg/kg i.p.), without modifying horizontal activity. Repeated pretreatment with SR 48692 (1 mg/kg x 5 days) markedly reduced locomotion and rearings after an acute cocaine challenge (day 1), whereas the lower dose of SR 48692 (0.1 mg/kg) had no effect. SR 48692 (1 mg/kg), given daily before cocaine, also decreased cocaine-induced rearing on day 2, but had no effect on the following drug challenges (days 3-10). One week after discontinuing repeated cocaine injections, SR 48692 blocked vertical, but not horizontal, activity induced by an acute cocaine challenge. Rats treated repeatedly with cocaine showed an enhanced behavioral response characterized by the development of stereotypes, which were unaffected by SR 48692. Finally, treatment with SR 48692 did not alter corticosterone circadian secretion nor cocaine-stimulated corticosterone levels, indicating that the attenuation of the behavioral effects of cocaine after NT receptor blockade is not associated with blunted glucocorticoid secretion. These results indicate that administration of SR 48692 attenuates the locomotion and rearing response to cocaine but fails to modify stereotyped behavior, suggesting that SR 48692 modulates the behavioral effects of psychostimulant drugs by acting selectively on the mesolimbic dopaminergic system.
- Published
- 1998
- Full Text
- View/download PDF
42. Steroid effects on brain functions: an example of the action of glucocorticoids on central dopaminergic and neurotensinergic systems.
- Author
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Rostène W, Sarrieau A, Nicot A, Scarceriaux V, Betancur C, Gully D, Meaney M, Rowe W, De Kloet R, and Pelaprat D
- Subjects
- Adrenocorticotropic Hormone metabolism, Aging physiology, Animals, Cell Death, Corticosterone metabolism, Disease Models, Animal, Hypothalamus drug effects, Rats, Time Factors, Brain drug effects, Dopamine metabolism, Glucocorticoids pharmacology, Neurotensin metabolism
- Abstract
It is now clearly established that steroid hormones released from peripheral endocrine glands may, through specific receptors in the brain, directly regulate brain function. These effects may be rapid or involve long-term modifications at the genomic level. Concerning the glucocorticoids, their receptors are found in most neuronal cells, an observation which can be related to their widespread effects on neuronal metabolism. Furthermore, glucocorticoids are often related to stress. We have previously demonstrated that neonatal handling of the rat prevented excessive endocrine response to stress. In adults, this action appeared to protect the animal from potential damaging effects of glucocorticoids and from related impairment of cognitive functions. The effects of glucocorticoids are thought to involve an interaction of several central neurotransmitter systems. One such neurotransmitter is neurotensin, a neuropeptide which was reported to be closely related to central dopaminergic system regulation. This paper presents a rapid overview of the central effects of glucocorticoids and possible evidence for the interrelationship between these steroids, dopamine and neurotensin systems in the regulation of the hypothalamo-pituitary-adrenal axis. It provides a new way to approach stress responses and to develop new substances that may become potential drugs in the treatment of some psychiatric disorders.
- Published
- 1995
43. Effect of brain lesions on [3H]ohmefentanyl binding site densities in the rat striatum and substantia nigra.
- Author
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Masuo Y, Wang H, Pélaprat D, Chi ZQ, and Rostène W
- Subjects
- Animals, Enkephalin, Ala(2)-MePhe(4)-Gly(5)-, Enkephalins metabolism, Fentanyl metabolism, Male, Rats, Rats, Wistar, Brain Injuries metabolism, Corpus Striatum metabolism, Fentanyl analogs & derivatives, Receptors, Opioid metabolism, Substantia Nigra metabolism
- Abstract
We have recently demonstrated that [3H]ohmefentanyl, a non-peptidergic opioid ligand which was suggested to cross the blood brain barrier in contrast to other peptidergic opioid ligands, bound not only to mu opioid receptor sites but also to sigma sites. In order to examine whether [3H]ohmefentanyl can be used as a marker for mu sites, we investigated the effects of brain lesions on [3H]ohmefentanyl binding site densities, as compared with [3H][D-Ala2, MePhe4, Gly-ol5]enkephalin ([3H]DAGO), a selective mu ligand. These binding site densities were measured by quantitative autoradiography in the rat striatum and substantia nigra, two brain structures known to contain a high density of mu receptors, following lesions of the nigro-striatal dopaminergic pathway and striatal intrinsic neurons. Following unilateral nigral lesion with 6-hydroxydopamine, [3H]ohmefentanyl binding site densities were decreased in the patches (-35%) and matrix (-20%) of the ipsilateral striatum and in the lesioned substantia nigra pars compacta (-49%). Unilateral striatal lesion with quinolinic acid induced 72%, 61% and 50% decreases in [3H]ohmefentanyl binding in the patches and matrix of the lesioned striatum and in the ipsilateral substantia nigra pars reticulata, respectively. Similar results were obtained in the binding of [3H]DAGO. Indeed, a significant linear correlation was observed between [3H]ohmefentanyl and [3H]DAGO binding site densities. Therefore, mu opioid receptors may be mainly located on intrinsic neurons in the striatum, dopaminergic cell bodies in the substantia nigra pars compacta and nerve terminals of striatal efferents in the substantia nigra pars reticulata.(ABSTRACT TRUNCATED AT 250 WORDS)
- Published
- 1992
- Full Text
- View/download PDF
44. Mesencephalic dopaminergic neurons in primary cultures express functional neurotensin receptors.
- Author
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Brouard A, Pelaprat D, Dana C, Vial M, Lhiaubet AM, and Rostène W
- Subjects
- Animals, Autoradiography, Binding, Competitive, Cells, Cultured, Immunohistochemistry, Mesencephalon cytology, Neurotensin pharmacology, Rats, Rats, Inbred Strains, Receptors, Neurotensin, Tyrosine 3-Monooxygenase metabolism, Dopamine metabolism, Mesencephalon metabolism, Neurons metabolism, Receptors, Neurotransmitter metabolism
- Abstract
The cellular distribution and functional aspects of neurotensin (NT) binding sites in rat mesencephalic cells in primary culture were investigated by an original approach combining anatomical and biochemical studies. Using a double-labeling protocol combining 125I-NT receptor radioautography and tyrosine hydroxylase (TH) immunocytochemistry, we obtained the first direct visualization of NT binding sites on TH-immunoreactive neurons. Eighty percent of the TH neurons were endowed with NT binding sites, which can be observed on both cell bodies and processes. TH-immunoreactive neurons were characterized as dopaminergic neurons by their ability to take up dopamine in a benztropine- and nomifensine-sensitive manner. In the mesencephalic cultures, NT increased potassium-evoked release of tritiated dopamine, and the relative potencies of various NT-related peptides to increase dopamine release were in good agreement with their abilities to bind to NT sites. These results show for the first time that cultured rat mesencephalic dopaminergic cells express functional NT receptors. Finally, the specificity and distribution of NT receptors on dopaminergic neurons in primary culture are quite similar to what was observed in the adult rat brain using pharmacological and radioautographic approaches. These data indicate that NT can influence the activity of dopaminergic neurons at very early stages of the rat brain development.
- Published
- 1992
45. Early spontaneous deficiency of calcitonin renal binding sites in rats with a high incidence of calcitonin-secreting tumors (WAG/Rij).
- Author
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Bouizar Z, Rostène WH, Treilhou-Lahille F, Pidoux E, Milhaud G, and Moukhtar MS
- Subjects
- Aging, Animals, Autoradiography, Kidney Cortex metabolism, Kidney Medulla metabolism, Rats, Receptors, Calcitonin, Calcitonin metabolism, Carcinoma metabolism, Kidney metabolism, Rats, Inbred Strains physiology, Receptors, Cell Surface metabolism, Thyroid Neoplasms metabolism
- Abstract
Old rats of the WAG/Rij strain have a high incidence (50%) of medullary thyroid carcinoma, a calcitonin (CT)-secreting tumor. We have characterized and quantified the topographical distribution of [125I]salmon calcitonin (sCT) binding sites in the kidneys of this strain, as compared to Wistar CF rats (2% incidence of spontaneous medullary thyroid carcinoma). We report here that, up to 15 days of postnatal development, the distribution of CT-binding sites in the kidney of the WAG/Rij strain was quite similar to that found in developing and adult Wistar CF rats. However, from the age of 1 month, sCT-binding sites were dramatically reduced in both the medulla and the inner part of the kidney cortex, though plasma CT levels were not significantly different in both strains. Adult WAG/Rij rats bearing a transplanted tumor for 12 weeks had a high level of plasma calcitonin and exhibited an even greater reduction of both medullary and cortical sCT-binding sites. These results suggest that the modification in the CT-binding sites in WAG/Rij rats is not a consequence of a possible down regulation due to elevated circulating hormonal level but could be inherited and possibly associated with the later development of the tumor in this strain.
- Published
- 1987
46. Characterization and quantitative topographical distribution of salmon calcitonin-binding sites in rat kidney sections.
- Author
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Bouizar Z, Rostène WH, Moukhtar MS, and Milhaud G
- Subjects
- Animals, Binding, Competitive, Humans, Male, Rats, Rats, Inbred Strains, Receptors, Calcitonin, Salmon, Species Specificity, Swine, Kidney analysis, Receptors, Cell Surface analysis
- Abstract
Renal binding sites for labelled salmon calcitonin (sCT) were studied using cryostat sections and autoradiography. Increasing concentrations of unlabelled sCT inhibited 125I-sCT binding. 125I-sCT bound to a single site with a Kd of 2 nM and a number of sites of 220 fmol/mg protein. Mammalian calcitonins had low affinities and peptides unrelated to CT were devoid of any significant affinity for 125I-sCT receptors. Autoradiograms disclosed a high concentration of 125I-sCT receptors mainly located in the outer medulla and heterogeneously in the renal cortex. The distribution of specific binding sites is in agreement with the current concepts of renal action of calcitonin.
- Published
- 1986
- Full Text
- View/download PDF
47. Electron microscopic localization of neurotensin binding sites in the midbrain tegmentum of the rat. I. Ventral tegmental area and the interfascicular nucleus.
- Author
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Dana C, Vial M, Leonard K, Beauregard A, Kitabgi P, Vincent JP, Rostène W, and Beaudet A
- Subjects
- Animals, Autoradiography, Binding Sites, Male, Microscopy, Electron, Rats, Rats, Inbred Strains, Tegmentum Mesencephali ultrastructure, Neurotensin metabolism, Tegmentum Mesencephali metabolism
- Abstract
The distribution of specifically labeled neurotensin (NT) binding sites was examined by light and electron microscopic radioautography in the ventral tegmental area (VTA) and nucleus interfascicularis of the rat following incubation of lightly prefixed midbrain slices with the monoiodinated ligand, 125I-(Tyr3)-NT. Film radioautograms of whole 125I-NT-incubated slices exhibited intense NT displaceable binding throughout the VTA and interfascicular nucleus. In light microscopic radioautographs from 1-microns-thick sections taken from the surface of the slices, the label was found to be present both inside and outside neuronal perikarya. Probability circle analysis of silver grain distribution in electron microscopic radioautographs confirmed that a significant proportion (greater than 20%) of the specifically labeled binding sites was intraneuronal. The frequent association of these sites with profiles of rough endoplasmic reticulum or Golgi apparatus suggested that they corresponded in part to receptors undergoing synthesis and/or glycosylation. The remainder was associated with neuronal and/or glial plasma membranes, as attested by comparing the distribution of grains overlying apposed cellular elements with the distribution of hypothetical grains originating from randomly distributed membrane bound radioactive sources. Although the resolution of the technique did not make it possible to ascribe labeled membrane-bound receptors to either one of the apposed plasma membranes, their frequent association with interfaces involving the plasmalemma of perikarya and dendrites, together with the occurrence of silver grain alignments along the membrane of certain somata and dendrites suggested that a proportion of them was associated with the perikarya and dendrites of a subpopulation of ventral tegmental neurons. Interestingly, these perikaryal and dendritic receptors were not exclusively present on, or even concentrated opposite, abutting axon terminals but instead were more or less evenly distributed along the plasma membrane. Only an exceedingly small proportion was found to be associated with synaptic junctions. Such a low incidence makes it unlikely that only the synapse-linked binding sites correspond to functional receptors. On the contrary, the dispersion of labeled receptors seen here along the plasma membrane of presumptive dopamine neurons suggests that NT acts mainly in a paracrine or parasynaptic fashion in the ventral midbrain tegmentum.
- Published
- 1989
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