Doctoral School Days 2015 ABSTRACT BOOK

February 26 - 27th 2015

300, Boulevard Sébastien Brant, 67400 Illkirch ESBS - Pôle API Illkirch

Organizing team:

Charlotte Canteloup, Kevin Dorgans, Hajer El Oussini, Elea Heberle & Elise Savier --- PhD Students

Mélanie Muser & Serge Potier --- European Doctoral College

Luc Dupuis --- Research Director, INSERM U1118

 

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DOCTORAL SCHOOL DAYS 2015 : PROGRAM

Day1 : Thursday February, 26th 8 :30 Registration 9 :00 Presentation of the Doctoral School - Serge Potier, Head of the Doctoral School

9 :30 Oral & Blitz session 1 O1 : Mechanisms of pair formations and seperation in rooks (Corvus frugilegus)

- Palmyre Boucherie 9 :45 B1 : Study of a unique tRNA snatcher and its role in Plasmodium development

- Delphine Kapps 9 :48 O2 : The LRP1/Wnt5a signaling pathway protects against cellular cholesterol

accumulation - Marion Jenty

10 :03 B2 : Variants of androgen receptors in prostatic cancer microenvironments - Edwige Schreyer

10 :06 O3 : What is the cause of weight loss in Amyotrophic Lateral Sclerosis? - Pauline Vercruysse

10 :21 B3 : An impaired coherence of the life story narrative of patients with schizophrenia - Mélissa Allé

10 :30 Coffee break & Poster session 1 11 :00 Oral & Blitz session 2 O4 : Constitutively active androgen receptor variants upregulate expression of

mesenchymal markers in prostate cancer cells - Félicie Cottard

11 :15 B4 : Study of Myotubularin, a phosphatase responsible for centronuclear myopathy by using as a model the yeast Saccharomyces cerevisiae. - Myriam Sanjuan-Vazquez

11 :18 O5 : Modelling infection transmission in primate networks to predict centrality-based risk - Valéria Romano

11 :33 B5 : Activation patterns of the rat dorsal striatum and hippocampus depend on the spatial demand of navigation tasks in the double-H maze - Julien Gasser

11 :38 O6 : RFRP-3 and kisspeptin as new regulators of energy balance: lessons from the desert, Jerboa (Jaculus orientalis) - Rajae Talbi

11 :53 B6 : Effects of a free-choice high-fat high-sugar diet on circadian brain activity in mice - Aurea Blancas-Velazquez

12 :00 Lunch 13 :30 Inspiring Talk : Thomas Ebbesen 14 :30 Presentation of associations : DoctoNeuro, Addal & Espace Avenir 15 :00 Forum, Gallery of PhD’s & Scientific speed dating 17 :30 Coffee break & Poster session 2 18 :00 Social event : Les improbables – Théâtre de l’Oignon 19 :30 Dinner – La pause Quinoa

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Day2 : Friday February, 27th 9 :00 Welcoming of the participants

-Dr. Christophe Romier (President of Strasbourg Biology Society) 9 : 20 Conférence  plénière  «  Contre  tous  les  dogmes  et  algorithmes,  émergence  d’une  

stratégie  thérapeutique  innovante  pour  traiter  le  lupus  »  -­‐Pr.  Sylviane  Muller

10 :20 Candidates  talks  

12 :00 Jury Deliberation  13 :00 Lunch & SBS Award 14 :00 Oral & Blitz session 3 O7 : Subthreshold Information Decoding at the Cerebellar Granule Cell to

Purkinje Cell Synapse - Anais Grangeray Vilmint

14 :15 B7 : On the tracks of Theory of Mind in monkeys: attention, perception and perception and intention reading abilities - Charlotte Canteloup

14 :18 O8 : CD41 (Itga2b) identifies a novel lymphatic endothelial subset - Mélanie Chypre

14 :33 B8 : Understanding the Modular Information Processing in the Cerebellar Cortex for the Compensation of Dystonic Disorders in Mice - Orkan Ozcan

14 :48 O9 : Antibiofilmogram, a potential tool to optimize antibiotic therapy against bacteria forming biofilms in cystic fibrosis patients - Elodie Olivares

15 :00

Round Table « Life after a PhD » animated by Jérôme Grenêche (Scientific

Journalist) Guests :

Domitille Boudard (Neurotime) Hervé Cadiou (MCU UDS)

Alexandre Charlet (CR CNRS) Judith Eschbach (Inoviem)

Marie Pelé (Liberal Ethologist) Renaud Wagner (CNRS)

16 :30 Coffee break & Poster session 3 18 :00 Election of the best blitz, talk and poster

Prize distribution

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ORAL & BLITZ PRESENTATIONS  

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O1 - Mechanisms of pair formations and separations in rooks (Corvus frugilegus) Palmyre Boucherie, Céline Bret, Valérie Dufour Département d’Ecologie Physiologie et Ethologie, Institut Pluridisciplinaire Hubert Curien UMR 7178, CNRS, Strasbourg, France Keywords: rooks (Corvus frugilegus), relationship quality, pair formation, separation In social corvids like rooks (Corvus frugilegus), the reproductive pair is the core unit of the social structure, but all year long, they live in colonies where they can interact with individuals outside the pair. Many studies have focused on the relationship strength of the pair in juveniles groups. But very little is known about the mechanism underlying the establishment of a new relationship and its durability in adults. We investigated the formation of a new relationship in 4 pairs of captive adult rooks, following also the separation process in 3 other pairs. To do so, we first introduced new members into an existing captive colony of rooks (6 new birds in a group of 9). We used social network analysis to investigate the role of each individual in the establishment of new relationships and the disappearance of the pre-existing one whenever it occurred. We recorded all social interactions emitted and received by individuals, and all dyads of individuals in proximity as a measure of associations. We found that the pair formation is a gradual phenomenon, leading to behaviors and associations exclusive to the mated pair (close proximity, feeding, preening). Separations did occur, including among birds that had been paired for years. The separations occurred concomitantly with the formation of new pairs, the new partner was chosen among familiar individuals, and a transitory period of triad configuration (that included the former couple and the new partner) was observed. These results underline the importance of extra-pair relationship and their potential influence on the strength and the consistency of a mated pair in these birds.

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B1 - Study of a unique tRNA snatcher and its role in Plasmodium development Delphine Kapps1, Nassira Mahmoudi-Kaïdi1, Robert Ménard2, Magali Frugier1 1Architecture et Réactivité de l’ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, 67084 Strasbourg Cedex, France 2Unité Biologie et Génétique du Paludisme, Institut Pasteur 25-28 rue du docteur Roux, 75015 Paris, France Keywords: malaria, Plasmodium sporozoites, tRNA-binding protein, exogenous tRNAs import, blood stage Plasmodium, the parasite responsible for malaria, is mainly intracellular and has a complex life cycle within the mosquito vector and its vertebrate host. The laboratory identified a surface protein expressed all along the life cycle of Plasmodium parasites. First, we chose to work with a particular stage, called the sporozoite. We demonstrated that this protein, called tRip for tRNA import protein, displays a tRNA binding domain outside of the parasite and mediates the import of exogenous tRNAs into sporozoite’s cytosol. Yet, we do not know the role of these tRNAs once inside the parasite. My PhD project begun with the design and the construction of a P. berghei parasite Knock Out for tRip. tRip-KO sporozoites are unable to import exogenous tRNAs, highlighting the need for this protein in this process. Since tRNAs are key molecules in translation, we looked for de novo protein synthesis in KO and WT sporozoïtes and showed that the global proteome seems to be disturbed in KO sporozoites compared to WT. Concerning in vivo experiments, tRip-KO clones developed normally inside mosquitoes and induced a normal liver stage in mice. However, blood infection, occurring after the liver stage, was abnormally low in KO-parasites-infected mice. All together, these experiments show that the import of host’s tRNAs has an impact on Plasmodium biology and is useful for its development. Further studies including mass spectrometry analysis should give us insightful information about the precise role that imported tRNAs could play in this parasite.

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O2 - The LRP1/Wnt5a signaling pathway protects against cellular cholesterol accumulation M. Jenty ,El Asmar Z, V. Bruban-Schann, J. Terrand, R. Matz-Westphal, L. Host, Justiniano H, Boudier C, P. Boucher Equipe Physiopathologie des maladies métaboliques, UMR 7213 LBP, Faculté de Pharmacie, Illkirch Atherosclerosis is characterized by the infiltration of foam cells laden with cholesterol in the arterial wall. We previously showed that the Low Density Lipoprotein Receptor Related Protein 1 (LRP1) protects from atherosclerosis. Indeed, mice lacking LRP1 specifically in vSMC develop atherosclerosis with massive foam cell formation. We also reported that LRP1 prevents cholesterol intracellular accumulation through a Wnt5a signaling pathway, however, how the LRP1/Wnt5a pathway regulates cholesterol homeostasis is unknown. Here we show that through LRP1, TGFβ induces Wnt5a expression. Wnt5a further inhibits cholesterol biosynthesis and increases cholesterol efflux. By interfering with the nuclear translocation of the transcription factor SREBP2 it down regulates HMGCoA-reductase and HMGCoA-synthase expressions, two keys enzymes of the cholesterol biosynthesis. Wnt5a also stimulates cholesterol export by induction of the ABCG1 transporter. We confirmed our in vitro data by generating mice overexpressing Wnt5a specifically in the adipose tissue. In white adipose tissue, Wnt5a decreases the expression of the HMGCoA-reductase and the HMGCoA-synthase by 90% in aTgWnt5a mice compared to controls, whereas ABCG1 expression is X15 in the mutant. Expression of Insig1, which retains SREBP2 in the ER and blocks its nuclear translocation, is increased by 6 fold. These results highlight new functions for LRP1 and the canonical Wnt5a pathway, and new therapeutic potential for atherosclerosis.

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B2 – Variants of androgen receptors in prostatic cancer microenvironments Edwige SCHREYER Team “Developmental and cell stress signaling in digestive and urological cancers” INSERM, Hopital civil, Strasbourg The receptor for androgens is a ligand-dependant transcription factor that plays a key role in the evolution of prostatic cancer. Evolved states of the illness are characterized by the presence of truncated variants of this receptor. Those variants show ligand-independent activation properties. Moreover, the core of the tumor also plays a key role in cancer progression especially through fibroblasts associated to cancer (CAFs). CAFSs can originate from mesenchymal stem cells (MSCs). My working hypothesis states that variants of the androgen receptors expressed in prostatic cancerous cells migh influence positively the differenciation of mesenchymal stem cells CAFs.

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O3 - What is the cause of weight loss in Amyotrophic Lateral Sclerosis? Pauline Vercruysse, Hajer EL OUSSINI, Jelena SCEKIC-ZAHIROVIC, Jerôme SINNIGER, Albert LUDOLPH, Anke WITTING, Luc DUPUIS and the GERP-ALS group INSERM U1118: Mécanismes centraux et périphériques de la neurodégénérescence, Faculté de médecine de Strasbourg Keywords: ALS, weight loss, hypothalamus Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease, occurring between 50 and 65 years old, characterized by a progressive paralysis and death within 3 to 5 years after diagnosis. ALS is also associated with defects in energy homeostasis in particular weight loss and hypermetabolism occurring in patients, but also in animal models of the disease. The objective of my PhD is to investigate the cause of weight loss in ALS. Body weight is mainly controlled by the hypothalamus, a brain structure which tunes the balance between food intake and energy expenditure. Interestingly, pioglitazone, a drug known to increase weight through hypothalamus, did not increase body weight in patients. Mouse models of ALS displayed abnormal food intake behaviors in response to either pioglitazone or fasting. Moreover, we observed ALS-related protein aggregates in the lateral hypothalamic area of patients and mouse models. To understand how hypothalamic alterations might alter energy homeostasis in ALS, we measured gene expression of 12 hypothalamic neuropeptides. Three were differentially expressed, with only MCH expressed in the lateral hypothalamus. Additionally, we observed decreased numbers of MCH positive neurons of ALS mice, and obtained preliminary evidence of decreased MCH levels in CSF of ALS patients. To determine whether loss of MCH was responsible for weight loss, we implanted ALS and control mice with osmotic mini-pumps delivering MCH during two weeks. MCH supplementation reverted weight loss in these mice. Our studies suggest that the loss of MCH could be a contributor to weight loss in ALS patients, and suggest ways to therapeutically correct weight in ALS patients.

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B3 - An impaired coherence of the life story narrative of patients with schizophrenia Mélissa Allé, Jevita Potheegadoo, Christin Köber, Priscille Schneider, Romain Coutelle, Tilmann Habermas, Jean-Marie Danion, Fabrice Berna INSERM U-1114, 1 place de l’Hôpital, Clinique Psychiatrique, Strasbourg Cedex, France Keywords: life story, schizophrenia, autobiographical reasoning, executive functioning, autobiographical memory Self-narratives of patients have received increasing interest in schizophrenia since they offer material enabling to figure out the experience of patients during the course of their illness, in particular the alteration of self-accompanying schizophrenia. In this study, we aimed at investigating the life story narrative of patients with schizophrenia and their ability to integrate and bind memories of personal events into a coherent narrative of their whole life. Twenty-seven outpatients and 26 controls were asked to recall the seven most important events they have lived and to integrate them in their life story narration. Two narrative components were analyzed: the narrative framework reflected by cultural and temporal aspects and autobiographical reasoning reflected by causal and thematic coherences. Our results showed that the cultural part of the narrative framework is preserved in patients whereas its temporal part is partially impaired. Furthermore, autobiographical reasoning is significantly decreased all along patients’ narratives: patients have difficulties to explain how lived events have modeled their identity and to integrate different lived events along thematic lines. Decrease of autobiographical reasoning was significantly correlated with patients’ executive dysfunction, suggesting that cognitive impairment observed in patients could affect their ability to construct a coherent narrative of their life binding important events to their self. This study provides new understanding of the cognitive deficits underlying self-disorders of patients with schizophrenia. Finally, patients with higher autobiographical reasoning skills reported stronger sense of coherence in their life, arguing for the development of therapeutic interventions which aimed at improving autobiographical reasoning skills.

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O4 - Constitutively active androgen receptor variants upregulate expression of mesenchymal markers in prostate cancer cells Félicie Cottard1, Pauline Berthélémy1, Irène Asmane1, Eva Erdmann1, Jean-Emmanuel Kurtz1,2, Jocelyn Céraline1,2

1INSERM U1113, Team 3 « Cell signalling and communication in kidney and prostate cancer », University of Strasbourg, France 2Haematology and Oncology Unit, Strasbourg University Hospital, France Keywords: Androgen receptor, tumor progression, Epithelial Mesenchymal Transition, N-cadherin Androgen receptor (AR) signaling pathway remains the main target of novel therapeutics for castration-resistant prostate cancer (CRPC). However, constitutively active AR variants lacking the carboxy-terminal region in CRPC lead to therapy inefficacy. Moreover, recent studies suggest that AR variants are expressed in primary prostate tumors and may contribute to tumor progression. The aim of this study was to investigate the impact of AR variants on tumor progression. N-cadherin expression was analyzed in LNCaP cells overexpressing the wild type AR or a AR variant by qRT-PCR, Western blot and immunofluorescence. We showed here that N-cadherin expression was increased in the presence of constitutively active AR variants. Moreover, we have shown that constitutively active AR variants seem to bind Androgen Response Elements (ARE) in the intron 1 of N-cadherin to induce its expression. Furthermore, an upregulation of other mesenchymal markers expression such as VIMENTIN, SNAIL and ZEB1 was observed in the presence of AR variants. Nevertheless, these mesenchymal markers up-regulation were not associated with a decrease of E-cadherin expression. This co-expression between epithelial and mesenchymal markers leads us to think that AR variants are associated to partial Epithelial Mesenchymal Transition. Finally, to have a better understanding of mechanisms leading to mesenchymal markers expression in the presence of AR variants, we have performed a RNA-seq and mi-RNA-seq experiment in LNCaP cells overexpressing the wild type AR or a AR variant. Taken together, our findings will allow us to have a better understanding of the mode of action of AR variants in prostate cancer.

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B4 - Study of Myotubularin, a phosphatase responsible for centronuclear myopathy by using as a model the yeast Saccharomyces cerevisiae Myriam Sanjuan-Vazquez Dimitri Bertazzi, Johan-Owen De Craene, Bruno Rinaldi and Sylvie Friant Génétique Moléculaire, Génomique, Microbiologie (GMGM), CNRS-Université de Strasbourg, 21 rue Descartes 67084 Strasbourg Cedex Keywords: Myopathy, myotubularins, MTM1, phosphoinositides, PH-GRAM domain, yeast Centronuclear myopathy (CNM) is a genetic muscular disease characterized at the histological level by nuclei at the center of the myofibers instead of the periphery. Mutations in three genes (MTM1, DNM2 and BIN1) are associated with this pathology. MTM1 on the X chromosome affects males and codes for an enzyme called myotubularin. Myotubularin is a phosphoinositide lipid phosphatase. The cellular processes controlled by MTM1 at the muscular level are still unknown. MTM1 study in human cells is complicated due to the presence of 14 homologues. Thus, we used the unicellular eukaryotic yeast model Saccharomyces cerevisiae to study MTM1. Yeast has a similar organellar organization as human cells and encodes only 1 myotubularin, termed Ymr1 (yeast myotubularin related 1). To better understand the cellular function of MTM1, I am studying the two main domains of this protein: the PH-GRAM Nterminal lipid binding domain and the C-terminal catalytic phosphatase domain. The two domains were tagged with different fluorescent reporters (GFP and mCherry) and either expressed or co-expressed into yeast cells. I visualized by fluorescent microscopy their cellular effects and localization. I also tested two CNM patient mutants affected into the PH-GRAM domain. The results show that intermolecular interactions between the N-ter PH-GRAM and the C-ter catalytic domain are required to restore a catalytically active phosphatase.

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O5 - Modelling infection transmission in primate networks to predict centrality-based risk Valéria Romano1,2, Julie Duboscq1,2, Cécile Sarabian2,3,6, Elodie Thomas4,6, Cédric Sueur1,2,5 and Andrew J.J. MacIntosh6,7

1Centre National de la Recherche Scientifique, Département Ecologie, Physiologie et Ethologie, Strasbourg, France 2Université de Strasbourg, Institut Pluridisciplinaire Hubert Curien, Strasbourg, France 3Université de Rennes 1, Rennes, France 4Université François-Rabelais, Tours, France 5Unit of Social Ecology, CP231, Université libre de Bruxelles, Campus Plaine, Brussels, Belgium 6Kyoto University Primate Research Institute, Inuyama, Japan 7Kyoto University Wildlife Research Center, Kyoto, Japan Keywords: agent-based model, social relationship, wildlife epidemiology, parasitology Social structure can theoretically regulate disease risk by mediating exposure to pathogens via social contact or spatial proximity. Investigating the role of central individuals within a network may help to predict infectious agent transmission as well as to implement disease control strategies, but little is known about such dynamics in real primate networks. We combined social network analysis and a modelling approach to better understand transmission of a theoretical infectious agent in wild Japanese macaques living on the islands of Koshima and Yakushima, Japan. Individual identities as well as grooming networks were included in a Markov graph-based simulation. In this model, the probability that an individual will transmit an infectious agent depends on the strength of its relationships with other group members. We correlated (i) the mean latency to complete transmission, (ii) the percentage of subjects infected during a latency-constrained epidemic, (iii) the probability that an individual is infected first among all group members and (iv) each individual's mean rank in the chain of transmission with individual network centralities. Our results show that more central individuals transmit infections in a shorter amount of time and to more subjects but also become infected more quickly than less central individuals, indicating that social network properties can mediate pathogen flow. This suggests the importance of social networks in disease transmission and illustrates how network analysis and modelling can help in predicting epidemics in primates.

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B5 - Activation patterns of the rat dorsal striatum and hippocampus depend on the spatial demand of navigation tasks in the double-H maze Julien Gasser, Brigitte Cosquer, Anne-Laurence Boutillier, Jean-Christophe Cassel Laboratoire de Neurosciences Cognitives et Adaptatives, UMR 7364, Université de Strasbourg-CNRS, 12 rue Goethe, F-67000 Strasbourg, France Keywords: Striatum, hippocampus, procedural memory, spatial navigation, allocentric & egocentric strategies Dorsal hippocampus and striatum, which contribute to learning about/navigating in an environment, are linked to different memory systems: dorsal hippocampus deals with navigation based on memories for configuration of environmental cues, whereas dorsal striatum mediates procedural/egocentric navigation strategies based on automatisms. In an early stage of spatial learning, rats use their hippocampus, but with further training they shift to a less effortful expression of automatisms. We provide c-Fos evidence to this view. Rats were trained during 1, 6 or 14 days (4 trials/day) in the Double-H maze, a novel water escape task (Pol-Bodetto et al, 2011). Using constant start point and goal location, a 1st group could see all visual cues (lights on; group CHOICE) and reach the goal with no constraint on strategy. The 2nd group was trained in darkness and had no access to visual cues (PR). In the 3rd group (SP for “spatial”), rats were released from a different start point on each trial and could see environmental cues. Rats taken from the home cage (HC) were our controls. After a probe trial given 24h after the last training session (at day 2, 7 and 15), rats were killed for cfos immunohistochemistry. Acquisition curves were comparable in all trained rats. C -fos analyses showed a strategy- and training level-specific activity pattern in the dorsomedian, dorsolateral striatum and in dorsal hippocampus. We conclude on a specific role of striatum and hippocampus during integration and retrieval of procedural and spatial memory, respectively.

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O6 - RFRP-3 and kisspeptin as new regulators of energy balance: lessons from the desert, Jerboa (Jaculus orientalis) Talbi R1,2, Klosen P2, Laran-chich MP2, EL Ouezzani S1, Simonneaux V2. 1 Laboratory of Neuroendocrinology and Nutritional and Climatic Environment, University of Sidi Mohammed Ben Abdellah, FSDM, PO Box 1796, ATLAS-FES, Morocco 2 Institut des Neurosciences Cellulaires et Intégratives, UPR CNRS 3212, Laboratory of Neurobiology of Rhythms, 5 rue blaise pascal, 67034 Strasbourg, France Keywords: Kisspeptin, RFRP-3, POMC, Somatostatin, NPY, Jerboa, reproduction, energy balance Kisspeptin is a hypothalamic neuropeptide known as a central regulator of reproduction. The purpose of this study was to investigate the involvement of kisspeptin in the seasonal variations of energy balance in Jerboa, a long day breeding rodent sexually active in spring. We analyzed the effect of acute intracerebroventricular injections of kisspeptin on food intake during spring and autumn. The animals received kisspeptin or vehicle injections, and the food ingestion was measured for the next 5 hours. In parallel, similar injections were performed but animals were sacrificed 1h30 after the injections in order to analyze the gene expression of neuropeptides involved in the regulation of energy balance (NPY, POMC, and Somatostatin). In spring, the kisspeptin administration inhibited the early intake during the 1-3h period post injection and reduced the food intake for the next 2 hours. In contrast, kisspeptin had no effect on food intake in autumn, suggesting the existence of a seasonal effect of kisspeptin on food intake. Kisspeptin injection increased the gene expression of both POMC and somatostatin in the two seasons but had no effect on the expression of NPY encoding gene. In contrast, kisspeptin displayed a differential effect on the RFRP-3 encoding gene, increasing its expression in autumn but decreasing it in spring. These results indicate that kisspeptin regulate Jerboa’s food intake, and its effect may involve a seasonal dependent action on RFRP3 expression. This work reveals the strong involvement of kisspeptin in the regulation of two crucial systems ensuring body homeostasis, reproduction and energy balance.

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B6 - Effects of a free-choice high-fat high-sugar diet on circadian brain activity in mice Blancas-Velazquez Aurea1,2, la Fleur Susanne2, Mendoza Jorge1

1Institute des Neurosciences Cellulaires et Intégratives, University of Strasbourg (UNISTRA) 2Academic Medical Center, University of Amsterdam (UvA) Rats under a normal chow food diet eat during their activity period in a daily rhythmic pattern. Interestingly, when they also have free access to fat and sugar, the intake of the pleasurable food is ingested equally during day and night, losing the circadian pattern. The aim of our work is to understand how the availability of highly rewarding diets alters the circadian rhythmicity of reward and/or metabolic brain structures leading to the aberrant daily food intake pattern. Methods: wild type C57BL6J male mice were randomly assigned to 2 groups: 1) a control group with normal chow-food and tap water ad libitum; 2) a free choice High-Fat High-Sugar (fcHFHS) diet, with choice among sugar (10% in tap water), fat (beef lard) vs. regular chow food and tap water provided ad libitum. 6 weeks after the exposure to these regimens, mice are sacrificed and brains dissected for imunohistochemical detection of c-Fos and the clock protein PER2 in the suprachiasmatic nucleus (SCN), hypothalamic (arcuate nuclei; ARC) and reward related areas (lateral habenula; LHb). In a second experiment under the same experimental conditions, using transgenic mice in which the protein PER2 is fused to the luciferase reporter, we follow, ex vivo, the circadian oscillations of PER2 in the brain of control and fcHFHS mice. Results: Mice with fcHFHS diet gained body weight and lost rhythmic feeding specifically of fat and sugar. SCN was not affected by the diet while ARC and LHb are disrupted.

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O7 - Subthreshold Information Decoding at the Cerebellar Granule Cell to Purkinje Cell Synapse Grangeray Vilmint A, Isope P CNRS UPR3212, INCI, Strasbourg, France Temporal coding in Purkinje cells (PCs), the sole output of the cerebellar cortex, plays a major role in motor control. Alteration of PC discharge leads to movement disorders such as ataxia. Each PC receives 170000 inputs from granule cells (GCs), the relay of the mossy fibers. GCs also provide a feedforward inhibition on PCs through activation of molecular layer interneurons. Since PCs are spontaneously active, we asked whether and how burst of subthreshold GCs inputs combined with molecular layer interneuron feedforward inhibition could modify PC discharge thereby influencing cerebellar cortex output. On acute cerebellar slices, burst stimulations (3 to 7 stims, 200 Hz) were elicited on a few GCs using extracellular stimulation and PC discharge was monitored in juxtacellular recordings. At the end of the experiment, PCs were whole-cell patched and both excitatory and inhibitory synaptic charges were determined. In order to mimic physiological conditions, we used Thy1-Channelrhodopsin2 mice combined with optogenetic illumination to reproduce “in vivo-like” level of synaptic background activity. We then analyzed the trains of spikes of PC and found that the structure of the train was correlated with the balance between excitation and inhibition. We also observed that the number of stimulations in the burst influences the net effect of GCs inputs on PC discharge. Our results demonstrate that the organization of local microcircuits and the temporal organization of GCs inputs may control the output of the cerebellar cortex.

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B7 - On the tracks of Theory of Mind in monkeys: attention, perception and intention reading abilities Charlotte Canteloup1,2, Hélène Meunier1,2 1Centre de Primatologie de l’Université de Strasbourg. 2Laboratoire de Neurosciences Cognitives et Adaptatives, UMR 7364, Université de Strasbourg The existence of the Theory of Mind - attribution of mental states to others, such as perspective-taking, intentions, desires or beliefs - in non-human primates, is historically a source of debate. Some scholars even put into question the actual existence of primates’ mental states reading abilities when others suggest that these cognitive abilities did not begin de novo but would rather be rooted in our primate heritage. The majority of research have been done on great apes and have reported discordant results, certainly due to experimental artifacts. The objective of my PhD is to investigate whether macaques and capuchins, species more distant from us than are great apes, share with us some Theory of Mind precursors, such as attention, perception and intention reading abilities. The three main questions of my PhD project are thus the following ones: Do monkeys are able to discriminate human attentional states? Do monkeys are capable of taking the visual perspective of conspecifics? Do monkeys are able to discriminate human intentions and prosocial versus antisocial behaviours of animated characters? Experiments planned will allow us to test subjects in different experimental contexts: in cooperation, competition and socially neutral context. Monkeys will be tested with human experimenters, conspecifics and, more innovative, in automatic computerized workstations equipped with eye-tracker. Studying Theory of Mind precursors in our closest relatives, the non-human primates, will help tracing back the evolutionary roots of human mindreading abilities and to shed new light on what being human means.

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O8 - CD41 (Itga2b) identifies a novel lymphatic endothelial subset O. Cordeiro1, M. Chypre1, S. Rauber1, C. Benezech2, N. Brouard3, F. Lanza3, Christopher G. Mueller1

1CNRS UPR3572, IBMC, University of Strasbourg, France 2Queens Medical Research Institute, University of Edinburgh, UK; 3UMRS 949 INSERM, Université de Strasbourg, EFS, France Keywords: CD41, Itga2b, Lymph node, Lymphatic endothelial cells A transcriptional profiling study of stromal cells identified CD41 (Itga2b/integrin aIIb) gene expression by lymph node lymphatic endothelial cells (LECs) (Malhotra, D. et al., 2012). Before this reference to CD41 expression by LECs, this integrin was thought to be restricted to hematopoietic stem cells in embryonic aorta and to platelets. This surprising finding attracted our attention. We therefore evaluated CD41 expression by lymph node (LN) stromal cells, including LECs. We also verified CD61 expression (Itgb3/integrin b3) since it is well known to form heterodimers with CD41 forming the IIb/IIIa glycoprotein complex (Du and Ginsberg, 1997). We found, by flow cytometry, that CD41 expression is mainly present on LECs and it appears as two heterogeneous populations, CD41hi and CD41low LECs. Regarding CD61 expression, we found it with different levels on the four stromal subsets (LECs, FRCS, BECs and DNCs), however, the heterodimer CD41/CD61 was identified using a complex-specific antibody only on LECs. Using confocal microscopy we could assess the localization of those cells within the LN. Again there was a heterogeneous distribution: on medullary LECs, cortical LECs and on the sucapsular sinus LECs. Strikingly of the two subcapsular sinus layers only the inner-sinus lining layer expressed CD41. We confirmed the expression of CD41 using a mouse model with GFP inserted into the cd41 gene. These finding shed new light onto LEC heterogeneity and provide novel means to target them for therapy or to better elucidate the role of LECs in LN homeostasis. Malhotra, D., Fletcher, A.L., Astarita, J., Lukacs-Kornek, V., Tayalia, P., Gonzalez, S.F., Elpek, K.G., Chang, S.K., Knoblich, K., Hemler, M.E., et al. (2012). Transcriptional profiling of stroma from inflamed and resting lymph nodes defines immunological hallmarks. Nat Immunol 2012, 2262.

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B8 - Understanding the Modular Information Processing in the Cerebellar Cortex for the Compensation of Dystonic Disorders in Mice Orçun Orkan Özcan, Philippe Isope Institut des Neurosciences Cellulaires et Intégratives, Centre National de la Recherche Scientifique, Unité Propre de Recherche 3212, Université de Strasbourg, 67084 Strasbourg, France Keywords: sensori-motor integration, motor coordination, cerebellum, dystonia The cerebellum plays a central role in sensori-motor integration in vertebrates. Increasing evidence indicate the involvement of the cerebellum in a large spectrum of movement disorders: ataxia, dystonia, essential tremor and Parkinson's tremor. Furthermore, direct stimulations of the cerebellum improve the condition and motor performances of patients suffering from cerebral palsy. The cerebellar system is organized in a series of parallel anatomical modules which govern its functional interactions with the rest of the sensorimotor system. This regionalization may be particularly relevant to the pathophysiology of focal dystonia. An association between cerebellar alterations and focal dystonia was noted by Fletcher et al. (1988). However, very little effort has been made so far to relate the modular organization of the cerebellar cortex and its relevance to cerebello-cortical interactions in dystonia. The various experimental models of dystonia consistently indicated a pathological activation of the cerebellum (Pizoli et al., 2002; Neychev et al., 2008). Therefore, documenting and modeling the functional connectivity in cerebellar modules could help to explain the focal nature of abnormal activity in experimental models of dystonia. In this presentation, we will explain an ongoing project that uses electrophysiological recordings (in vivo), advanced data analysis and modeling of network dynamics, both in individual modules and at the output stage of the cerebellum. The study of individual modules and their influence on the output of the cerebellar cortex in normal and pathological condition will allow us to develop strategies of cerebellar stimulation in vivo that could compensate dysfunctions observed in models of dystonia.

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O9 - Antibiofilmogram, a potential tool to optimize antibiotic therapy against bacteria forming biofilms in cystic fibrosis patients Elodie Olivares1,2, Stéphanie Badel-Berchoux1, Christian Provot1, Thierry Bernardi1, Gilles Prévost2, François Jehl2 1BioFilm Control, Saint Beauzire, France 2Institut de Bactériologie, EA7290 Virulence bactérienne précoce, Strasbourg, France Keywords: biofilm, Pseudomonas aeruginosa, cystic fibrosis, antibiotics, Biofilm Ring Test, antibiofilmogram Bacteria, as others unicellular micro-organisms, can adopt two different ways of life: the planktonic mode, where bacteria are in a free floating state in a liquid environment, or the sessile mode, which corresponds to the biofilm. This structural organisation is defined as a multi-cellular community of micro-organisms, which adhere to a surface and is characterized by the secretion of a protective matrix. Clinically, 60% of infections are due to bacteria organized in biofilm, including a lot of pathologic situations: infective endocarditis, urinary and osteoarticular infections, cystic fibrosis... Their treatment is based on the realisation of standard antibiograms to evaluate the bacterial susceptibility to antibiotics. These routinely used tests are carried out on planktonic bacteria. It is well established that sessile micro-organisms susceptibility to antibiotics differs from their planktonic phenotype. The development of a new technology, the Biofilm Ring Test, allows biofilm formation monitoring. Thus, therapeutic molecules may be investigated as regards their preventive and/or curative impact on biofilms formation (antibiofilmogram term is also used). We tested many antibiotics for their ability to prevent biofilms formation by a collection of Pseudomonas aeruginosa strains, isolated from cystic fibrosis patients. Results indicate that the responses of bacteria to antibiotics are frequently varying, depending on the strains phenotype and when compared to data documented by classic antibiograms. Some antibiotics are able to induce the formation of a biofilm by bacteria. Therefore, antibiofilmogram appears to be a promising tool to take into account the antibiotics activity on sessile bacteria and could be essential for the therapeutic choice, in the long term.

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POSTER PRESENTATIONS  

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P1 - Impacts of the cellular factor CTIP2 on HIV-1 latency Involvement on the establishment and the persistence of the viral reservoirs Amina Ait Ammar, Christian Schwartz, Olivier Rohr Université de Strasbourg, EA7292, DHPI, Institut de Parasitologie et pathologies tropicales, 3 rue Koeberlé, 67000 Strasbourg France Keywords: HIV-1 transcriptional latency, p-TEFb, CTIP2, Tat The HIV-1 post-integrative latency generates reservoirs which prevent the eradication of the virus with the current therapies. We have demonstrated that the transcriptional co-repressor CTIP2 recruits multi-enzymatic complexes to promote the establishment and the persistence of latency in microglial cells, the major HIV-1 reservoir in the central nervous system. The control of the HIV-1 Tat protein is crucial to the persistence of these long life reservoirs. Tat reactivates the latent viruses by recruiting the cellular elongation factor P-TEFb to the TAR region of nascent HIV-1 RNA. Recently, we reported that the CTIP2 associates and inhibits P-TEFb in an inactive nucleo-proteic complex containing the cellular 7SK snRNA. We believe that this function of CTIP2 favors the persistence of the latently infected cells. The goal of my PhD is to purify and characterize the CTIP2-associated complexes in the absence (non infected cells) and in the presence of the viral protein Tat (HIV-1 infected cells). We will use several biochemical approaches such as gel filtration and glycerol gradients to purify the CTIP2-associated complexes. In addition, combined RIP-seq and proteomic approaches will allow us to determine the nature of the associated RNAs and proteins respectively. Finally, we will investigate on the functional impacts of the discovered CTIP2- associated complexes on cell line models and primary HIV-1 target cells.This work is part of a global project aiming to define therapeutic targets for a HIV-1 functional cure.

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P2 - Circuit mediating inhibition of olfactory bulb periglomerular cells Álvaro Sanz Díez, Didier De Saint Jan INCI, CNRS UPR3212, Strasbourg, France Keywords: olfactory bulb, interneurons, microcircuits The olfactory bulb (OB) is the first relay station in the brain for odor processing. It receives sensory afferents from olfactory sensory neurons (OSN). This information is transmitted to mitral and tufted cells, the principal output neurons of the bulb, within anatomical structures called glomeruli. Each glomerulus is surrounded by periglomerular (PG) cells that mediate intraglomerular inhibition. We recently characterized a specific PG cell subtype that plays a dominant role in mediating OSN-evoked intraglomerular inhibition of mitral and tufted cells (Najac et al. submitted). This PG cell subtype that expresses EYFP in Kv3.1-EYFP transgenic mice represents 20-30% of all PG cells, is axonless and projects its dendrites into a single glomerulus. EYFP(+) PG cells, like most PG cells, receive spontaneous inhibitory synaptic inputs (IPSCs) at an average frequency of 1.17 ± 1.05 Hz (n=9). We have examined the circuit mediating this inhibition. Using whole-cell patch-clamp recordings in horizontal slices of OB from Kv3.1-EYFP transgenic mice, we found that stimulation of OSN that produce excitatory postsynaptic currents (EPSC) in EYFP(+) PG cells voltage-clamped at negative holding potentials (Vh = -75mV) did not evoke any IPSCs when the cells where clamped around the reversing potential for excitation (Vh = 0mV, n=10). In contrast, stimulation within distant (>200µm) glomeruli produced a gabazine-sensitive monosynaptic IPSC (average amplitude 58 ± 36 pA, n=10) resistant to NBQX and D-AP5. Evoked IPSC had 20-80% rise time (0.49 ± 0.12 ms) and half-width (14.9 ± 4ms) that were similar to those of spontaneous IPSCs (0.56 ± 0.08ms and 15.5 ± 2.63 ms, respectively). These results suggested that inhibitory inputs onto EYFP(+) PG cells are not generated by the glomerular network they are associated with but instead are provided by neurons with axons running within the glomerular layer. Thus, we suspect short-axon cells located in infra mitral cell layers (deep SA cells) or in the glomerular layer (superficial SA cells) that have an axon that ramifies across several glomeruli to be the neurons inhibiting EYFP(+) PG cells.

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P3 - Effect of a short-term personalized Intermittent Work Exercise Program (IWEP) on endurance parameters and blood pressure among healthy older seniors Walid Bouaziz1,2, Elise Schmitt1,2, Evelyne Lonsdorfer2,3, Georges Kaltenbach1, Bernard Geny2,3 and Thomas Vogel1,2 1 Geriatric department, University Hospital, Strasbourg, France. 2 Department of physiology and EA-3072, Faculty of Medicine, Strasbourg University, Strasbourg, France. 3 Functional Explorations Department, University Hospital, Strasbourg, France Keywords: IWEP, older seniors, endurance parameters, blood pressure. Introduction: Regular physical activity is essential for healthy aging. The purpose of this study is to investigate the effect of the IWEP on endurance parameters and blood pressure among seniors. Methods: Eighty-one patients over sixty years were enrolled and have benefited of IWEP. The IWEP program consists of a 36-min cycling workout twice a week, 36 sessions in all. The first ventilatory threshold (VT1), heart rate (HR) at pre-training VT1, total workload (TW), systolic (SBP) and diastolic blood-pressure (DBP) were measured before, after the 18th and the 36th sessions. Results: For men, VT1 and TW was improved after the 18th sessions by 25,7% and 19,1% and after the 36th sessions by 35,7% and 23,9%, respectively (z>1,65). For women, VT1 and TW was improved after the 18th sessions by 32,4% and 26,1% and after the 36th sessions by 45,2% and 32,9%, respectively (z>1,65). For the HR at pre-training VT1, there was a significant decrease after the first 18th sessions for men by 6,2% and for women by 6,1% (z>1,65).This decrease remains significant after the 36th sessions in both groups. For women, SBP and DBP was decreased significantly after the 18th sessions by 8,7 mmHg and 4,4 mmHg and after the 36th sessions by 8,7 mmHg and 3 mmHg, respectively (z>1,65). For men, only the SBP was significantly reduced after the 18th sessions by 4,8 mmHg (z>1,65). Conclusion: The IWEP training program has shown a great improvement on endurance parameters and blood pressure among healthy older seniors.

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P4 - Rhesus macaques are more sensitive to finer attentional cues than Tonkean macaques Charlotte Canteloup1,3, Dalila Bovet2 and Hélène Meunier1,3

1Centre de Primatologie de l’Université de Strasbourg 2Laboratoire d’Ethologie, Cognition et Développement de l’Université Paris Ouest Nanterre la Défense 3Laboratoire de Neurosciences Cognitives et Adaptatives, UMR 7364, Université de Strasbourg The present study asked whether two socially divergent macaques’ species, the little tolerant rhesus macaques (Macaca mulatta) and the more egalitarian Tonkean macaques (M. tonkeana) share with us attention reading abilities. More specifically, we investigated which attentional cues – body, face, eyes orientation – were taken into account by these species to communicate through a learned pointing gesture the location of an unreachable food reward to a human partner. Macaques were tested in seven experimental conditions differing according to the human partner’s attentional state. The experimenter had her 1) eyes open and looked at the macaque, or 2) eyes closed, or 3) eyes up, or 4) she did the same “gaze alternation” as the subject, or 5) she was back turned, or 6) she turned her head aside, or 7) she was absent. We hypothesized that rhesus macaques living in a highly hierarchical society, should be more sensitive to subtle social cues such as eyes state than Tonkean macaques undergoing fewer social pressures within their group. Our results showed that rhesus macaques discriminated more and finer cues than Tonkean macaques. While both species seem sensitive to the presence and the body orientation of the human partner, only rhesus macaques are sensitive to the face orientation of the human. Moreover, although particularly sensitive to the “alternation” condition, neither Tonkean macaques nor rhesus macaques discriminated the open/closed eyes state. However, rhesus monkeys showed more gaze alternation and therefore monitored more carefully the attentional state of the human partner than did Tonkean macaques.

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P5 - CD41 (Itga2b) identifies a novel lymphatic endothelial subset O. Cordeiro1, M. Chypre1, S. Rauber1, C. Benezech2, N. Brouard3, F. Lanza3, Christopher G. Mueller1

1CNRS UPR3572, IBMC, University of Strasbourg, France 2Queens Medical Research Institute, University of Edinburgh, UK 3UMRS 949 INSERM, Université de Strasbourg, EFS, France Keywords: CD41, Itga2b, Lymph node, Lymphatic endothelial cells A transcriptional profiling study of stromal cells identified CD41 (Itga2b/integrin aIIb) gene expression by lymph node lymphatic endothelial cells (LECs) (Malhotra, D. et al., 2012). Before this reference to CD41 expression by LECs, this integrin was thought to be restricted to hematopoietic stem cells in embryonic aorta and to platelets. This surprising finding attracted our attention. We therefore evaluated CD41 expression by lymph node (LN) stromal cells, including LECs. We also verified CD61 expression (Itgb3/integrin b3) since it is well known to form heterodimers with CD41 forming the IIb/IIIa glycoprotein complex (Du and Ginsberg, 1997). We found, by flow cytometry, that CD41 expression is mainly present on LECs and it appears as two heterogeneous populations, CD41hi and CD41low LECs. Regarding CD61 expression, we found it with different levels on the four stromal subsets (LECs, FRCS, BECs and DNCs), however, the heterodimer CD41/CD61 was identified using a complex-specific antibody only on LECs. Using confocal microscopy we could assess the localization of those cells within the LN. Again there was a heterogeneous distribution: on medullary LECs, cortical LECs and on the sucapsular sinus LECs. Strikingly of the two subcapsular sinus layers only the inner-sinus lining layer expressed CD41. We confirmed the expression of CD41 using a mouse model with GFP inserted into the cd41 gene. These finding shed new light onto LEC heterogeneity and provide novel means to target them for therapy or to better elucidate the role of LECs in LN homeostasis. Malhotra, D., Fletcher, A.L., Astarita, J., Lukacs-Kornek, V., Tayalia, P., Gonzalez, S.F., Elpek, K.G., Chang, S.K., Knoblich, K., Hemler, M.E., et al. (2012). Transcriptional profiling of stroma from inflamed and resting lymph nodes defines immunological hallmarks. Nat Immunol 2012, 2262.

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P6 - Are schizophrenic patients dyslexics? Curzietti Maxime, Bonnefond Anne, Vidailhet Pierre, Doignon-Camus Nadège INSERM U1114, Pôle de Psychiatrie-Hôpital Civil de Strasbourg, 1 place de l'Hôpital, Strasbourg, France Keywords: Schizophrenia, Dyslexia, Reading, N170 Reading has been proposed among the cognitive functions affected in schizophrenia. Studies reported that reading fluency, accuracy and phonological processing was impaired for patients relative to controls (Martinez et al., 2013; Revheim et al., 2006, 2014). A phonological deficit is widely reported to be the characteristic of dyslexia. Revheim et al. (2014) proposed that 70% of patients in their sample met criteria for dyslexia. The question whether or not patients suffering from schizophrenia exhibit the same characteristics than dyslexics is of high relevance, because a common etiology may underlie reading disorders associated with schizophrenia and dyslexia (Condray, 2005). A way to explore the reading characteristics of schizophrenic patients is to explore the N170 component. Results in normoreaders reported larger N170 amplitude for visual wordlike stimuli than for non-orthographic stimuli (i.e., symbols) and have been interpreted as visual expertise for print (Bentin et al., 1999). Recently we showed that N170 print tuning was impaired in dyslexics (Mahé et al., 2012), as no amplitude difference between words and symbols has been observed. Moreover, we found that poor readers matched in reading level with dyslexics displayed N170 tuning, suggesting that impaired N170 specialization is a hallmark of dyslexia (Mahé et al., 2013). In the present study, we explore the visual expertise for print (i.e., the N170 component) in schizophrenic patients and controls. ERPs were recorded during a lexical decision. If schizophrenic patients exhibit characteristics of dyslexia, no modulation of the N170 component should be observed. In contrast, if schizophrenic patients exhibit garden-variety reading impairment, they should show similar N170 tuning as control participants.

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P7 - Evaluation of interest of combined training in Multiple Sclerosis and accompaniment of patients in their practice of sport ZAENKER Pierre 1,2,5, FAVRET Fabrice1,2, ISNER-HOROBETI Marie-Eve2,3, LONSDORFER Eveline2,6, DE SEZE Jérôme4,5

1Université de Strasbourg, Faculté des sciences du sport, Strasbourg, France, 2Université de Strasbourg, EA3072 Mitochodrie, stress oxydant et protection musculaire, Strasbourg, France 3Université de Strasbourg, Service de Médecine Physique et de Réadaptation, Institut Universitaire de Réadaptation Clémenceau (IURC)-Strasbourg, France, 4Université de Strasbourg, Service de Neurologie, CHU-Strasbourg Hautepierre, France 5Université de Strasbourg, Centre d’Investigation Clinique-Strasbourg, France, 6Université de Strasbourg, Service de Physiologie et Explorations Fonctionnelles NHC, Hôpitaux Universitaires de Strasbourg Keywords : Multiple sclerosis, aérobic exercise, Strength training, Quality of life. Background: Most of the studies used a model of aerobic exercise training at moderate intensity. However, it has been shown that interval aerobic exercise associated with strength training may be more efficient in several pathologies. Objectives: The objectives were to determine whether exercise training (i.e. interval aerobic exercise and strength training) may improve: 1) aerobic and strength capacity; 2) quality of life 3) the autonomy to exercise. Methods: 13 MS patients have already completed the program. A measurement of maximum oxygen consumption and maximal muscle strength of right (RKE) and left knee extensors (LKE) at 90-180 and 240°/s were made before and after training. Exercise training included 1 aerobic session on cycle ergometer and 1 session of muscular strengthening during the first 4 weeks, then1 session in autonomy is added. Aerobic exercise started by a 10 min warm-up, then 5times 1 min at 90% of maximal heart rate following by 3 min of recovery at anaerobic threshold. Results: The muscles strength was increased by 18% on LKE and 9% on RKE at 180°/sec. And by 10% on LKE and 7% on RKE at 240°/sec. VO2peak as well as maximal aerobic power were improved following exercise training respectively by 10 and 8%. The SEP-59 self-questionnaire showed that Role limitation - emotional was enhanced by 13%, energy by 7% and emotional well-being by 10%. Conclusions: Combined exercise training 1) improved strength and VO2peak 2) enhanced emotional well-being 3) encouraged the resumption of physical activity in autonomy for patients.

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P8 - Spinal cholinergic interneurons: Their recruitment for modulation of nociception Dhanasak Dhanasobhon, Rémy Schlichter and Matilde Cordero-Erausquin Institut des Neurosciences Cellulaires et Intégratives, UPR3212 CNRS, Dept. Nociception et Douleur, 67084 Strasbourg, France Keywords: Spinal cord; Pain; Nociception; Acetylcholine; Electrophysiology Endogenous acetylcholine (ACh) is an important modulator of nociceptive sensory processing in the spinal cord. We recently described a sparse population of cholinergic interneurons in the spinal dorsal horn of rodents and primates and suggested that it is the main source of ACh at this level. We aimed at identifying the features that enable this sparse neuronal population to achieve a major control over pain processing by elucidating the interplay of dorsal horn cholinergic interneurons with the surrounding nociceptive network. In order to elucidate the inputs they received, spinal horizontal and parasagittal slices were made from transgenic mice with enhanced green fluorescent protein expression in cholinergic interneurons and whole-cell recordings were made from these fluorescent neurons. We recorded spontaneous excitatory (EPSC) and inhibitory (IPSC) post-synaptic currents, as well as the changes in their frequency induced by various drugs including capsaicin, menthol and allyl isothiocyanate. In addition, we electrically stimulated the primary afferent roots attached to the slice. Our data suggests that spinal dorsal horn cholinergic interneurons receive both EPSCs and IPSCs; the frequency of these currents were altered during treatment with capsaicin confirming that cholinergic neurons are an element of the spinal nociceptive network. These neurons also respond to the stimulation of primary afferents at non-nociceptive and nociceptive stimulation intensity. Our study therefore provides important insights into the recruitment of the spinal cholinergic system.

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P9 - Design of therapeutic aptamers imported into mitochondria of human cells. Ilya S. Dovydenko1,2, Mariya I. Meschaninova1, Yann Tonin2, Anne-Marie Heckel2, Ivan Tarasov2, Aliya G. Venyaminova1, Nina Entelis2. 1Labotarory of RNA Chemistry, Institute of Chemical Biology and Fundamental Medicine, SB RAS, Lavrentiev ave., 8, Novosibirsk 630090, Russia 2UMR 7156, Universite de Strasbourg/Centre National de la Recherche Scientifique (UdS/CNRS), 67084 Strasbourg, France. Defects in mitochondrial genome often cause neuromuscular pathologies, for which no efficient therapy has been developed. Because most of deleterious mitochondrial mutations are heteroplasmic (wild type and mutated mitochondrial DNA (mtDNA) coexist in the same cell), the shift in proportion between two types of molecules could restore mitochondrial functions. Recently, we developed mitochondrial RNA vectors that can address anti-replicative oligoribonucleotides into human mitochondria and thus reduce the level of mtDNA bearing a deletion [1]. Here, we applied this strategy to point mutations. We demonstrate that specifically designed RNA molecules containing determinants for mitochondrial import and 20-nucleotide sequence corresponding to the mutated region of mtDNA, are able to anneal selectively to the mutated mitochondrial genomes. After import into mitochondria of cultured human cells, these RNAs induced a decrease of the proportion of mtDNA bearing a pathogenic point mutation in the ND5 gene [2]. To increase the stability of anti-replicative molecules in cells, we synthesized oligoribonucleotides contained different modifications in nuclease sensitive motives. These modified molecules were able to anneal to mutant mtDNA and their lifetime was increased, but they did not cause any reproducible effect on the heteroplasmy level [3]. Another way to stabilize the shift of heteroplasmy consists in several consecutive cell transfections. To decrease the toxicity of transfection procedure, we synthesized oligonucleotides conjugated with cholesterol [4]. These molecules are able to penetrate into cultured cells without additional lypophilic agents, partially localized in mitochondria and alter the heteroplasmy in cells bearing a pathogenic mutation. [1] Comte et al. (2013) Nucleic Acids Research.[2] Tonin et al. (2014) J. Biol. Chem.[3] Tonin et al. (2014) Biochimie.[4] Petrova et al. (2012) Nucleic Acids Research. This work was supported by grants Labex MitoCross, LIA ARN-Mitocure, AFM, ANR and Suprachem.

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P10 - Viral counteractions against CTIP2 in HIV-1 permissive cells Forouzanfar F., Ali S., Schwartz C. and ROHR O. University of Strasbourg, EA7292 DHPI, Institute of Parasitology and Tropical Pathology of Strasbourg, France Keywords: VIH-1, Vpr, CTIP2, Ubiquitination Latently infected cells constitute major blocks to an HIV-1 eradication and a functional cure of the patients. We have previously reported that the cellular co-factor CTIP2 plays a key role in the establishment and persistence of HIV latency in microglial cells, the long life reservoirs of the virus in the central nervous system. By recruiting large enzymatic complexes at the viral promoter, CTIP2 silences HIV-1 gene transcription and disfavors the viral reactivation from the reservoirs. Here, we report that interferon treatments induce expression of CTIP2 at the mRNA and the protein levels suggesting that this factor may be part of the cellular response to viral infections. Usurping the host ubiquitination machinery to target undesirable host proteins is a common strategy used by retroviruses. We observed that replication of WT- but not Vpr-deleted HIV-1 reduced CTIP2 expression in productively infected cells. Vpr expression was correlated with low levels of CTIP2 and increased levels HIV-1 gene transcription. Finally, the abrogation of Vpr binding to the DCAF1-CUL4-DDB1 complex counteracted CTIP2 degradation. Taking together, these results suggest that Vpr recruits the ubiquitination machinery to induce CTIP2 degradation and to bypass an undesirable response of the HIV-1 infected cells.

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P11 - A new experimental device measuring cognitive performances in nonhuman primates living in social group Fizet Jonas1,3, Rimele Adam1, Pebayle Thierry2, Cassel Jean-Christophe3, Kelche Christian3, Meunier Hélène1,3 1Centre de Primatologie de l’Université de Strasbourg, Fort Foch – Chemin du fort, 67207 - Niederhausbergen, France 2CNRS, Centre d’Investigation Neurocognitive & Neurophysiologique, UMS 3489, 21 rue Becquerel, BP 20, 67087 - Strasbourg, France 3CNRS, Laboratoire de Neurosciences Cognitives & Adaptatives, UMR 7364, 4 rue Blaise Pascal, CS 90032, 67081 - Strasbourg, France Keywords: Cognitive functions, Touchscreen, Non human primate, Automated system, Computerized tasks Discoveries are highly dependent on technological advances. In neurosciences, behavioural studies are crucial in that they can measure in a non-invasive way the visible part of cognitive processes. Studying the evolution of subjects' behavioural phenotypes through specific neuropsychological tasks already demonstrates its efficiency. We propose here to use such approach thanks to a new automated experimental device. Nonhuman primates, able to practice different paradigms used in humans and to learn several tasks, represent a key model to study normal or altered operating functioning of cognitive functions as well as interactions between these functions. Our apparatus is designed to accurately measure cognitive performances in semi-free ranging monkeys living in social group thanks to operant conditioning only, which excludes both deprivation and physical constraints. Subjects have a free and ad libitum access to the device thanks to an automated identification system. Tests are provided via a touch-screen interface, as in humans. Other original features of our experimental device lie in that i) several paradigms can be learned in parallel by monkeys, ii) the difficulty of each task changes in real time according to individual performances and iii) the full experimental device operates autonomously. The procedures established are particularly in agreement with new ethical standards of animal experimentation. We are currently validating the simultaneous learning of several cognitive tasks targeting memory and attention. Such tool will be of particular interest in the study of mechanisms underlying cognitive processes and more particularly normal or pathological cognitive aging phenomena.

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P12 - Regulation of poly(ADP-ribose) glycohydrolase function by phosphorylation Eléa Héberlé1, Giuditta Illuzzi1, Jean-Christophe Amé1, Vincent Heyer2, Bernardo Reina-San Martin2, Françoise Dantzer1 and Valérie Schreiber1

1 Biotechnology and Cell Signalling, UMR7242 CNRS, Université de Strasbourg, IREBS, Laboratory of Excellence Medalis, Equipe Labellisée Ligue contre le Cancer, ESBS, 300 Blvd Sébastien Brant, CS 10413, 67412 Illkirch, France 2Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), INSERM U964 / CNRS UMR 7104 / Université de Strasbourg, 67404 Illkirch, France. Among post-translational modifications, poly(ADP-ribosyl)ation has emerged to be a crucial event in a wide range of processes from DNA damage signalling to regulation of chromatin structure and gene expression. While poly(ADP-ribosyl)ation of proteins occurs through the activity of a family of 17 proteins called the poly(ADP-ribose) polymerases (PARPs), the reversion of this modification is essentially driven by the degrading activity of poly(ADP-ribose) glycohydrolase (PARG). Although PARG is encoded by a single gene in the human genome, its regulation is finely tuned by several processes such as alternative splicing and translational re-initiation, generating at least five isoforms displaying various subcellular localisations and functions. Regulation of PARG function or activity by post-translational modifications has not been addressed so far. The present work aims at deciphering the regulation of PARG function by phosphorylation. We have observed that a kinase activity co-purifies with PARG from cell extracts, PARG being itself a substrate for this phosphorylating activity. Proteomic studies identified several protein kinases involved in DNA damage response as PARG partners. For one candidate, in vitro phosphorylation assays using recombinant proteins validated that PARG is a specific target for phosphorylation. Our current work is focused on the identification by mass spectrometry of the phosphorylation site(s) within PARG, to subsequently generate unphosphorylable or phosphomimetic PARG mutants. Next, we will evaluate the influence of PARG phosphorylation status on its in vitro enzymatic activity. Through the generation of stable cell lines expressing these PARG mutants, we will evaluate the involvement of PARG phosphorylation on its function in DNA repair. We will also examine which PARG isoform is regulated by this phosphorylation. We expect that our results will shed light on the regulation of PARG that is needed to tightly control the level of PAR produced in response to DNA damage to avoid its detrimental accumulation.

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P13 - Sex differences in the photoperiodic regulation of RFRP and its receptor GPR147 in the Syrian hamster. Jo B Henningsen1, Vincent-Joseph Poirel1, Jens D Mikkelsen2, Kazuyoshi Tsutsui3, Francois Gauer1 and Valerie Simonneaux1 1Institute of Cellular and Integrative Neurosciences, CNRS, University of Strasbourg, Strasbourg, France 2Neurobiology Research Unit, Rigshospitalet, Copenhagen University Hospital, Denmark 3Laboratory of Integrative Brain Sciences, Department of Biology, Waseda University, Tokyo, Japan. Photoperiodic signals are transduced into the rhythmic release of the pineal hormone melatonin, which synchronizes reproduction with season. RF-related peptide (RFRP) is considered to play a role in the seasonal regulation of reproduction and its expression in the mediobasal hypothalamus is strongly down-regulated by melatonin in short photoperiod (SP). RFRP-3 regulates reproductive activity depending on gender and species but whether RFRP-3 exerts its actions directly on GnRH-neurons remains unclear. In this study we aimed to investigate photoperiod-dependent variations in the RFRP system in male and female Syrian hamster. We found that the neuroanatomical distribution of RFRP-immunoreactive fibers and GPR147 mRNA are similar in male and female Syrian hamster. However, both RFRP-neuronal expression and GPR147 mRNA levels are significantly higher in female than in male. GPR147 is mainly expressed in the preoptic area, anteroventral-periventricular nucleus, suprachiasmatic nucleus, paraventricular nucleus, bed nucleus of stria terminalis, ventromedial hypothalamus, habenular nucleus and the arcuate nucleus. GPR147 mRNA levels as well as RFRP-neuronal expression is down-regulated in SP, however with a stronger decrease in female than in male. The density of RFRP-fiber projections was studied in areas expressing GnRH- (preoptic area) and kisspeptin-neurons (AVPV and ARC). Interestingly, RFRP-fiber density in the AVPV was found to be higher in females in SP. RFRP and its receptor GPR147 are regulated by photoperiod in both male and female Syrian hamster. However, our results suggest that the RFRP system is more sensitive in female with a distinct role in the AVPV, probably in regulation of the pre-ovulatory LH surge.

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P14 - Cognitive control mechanisms involved in a sustained attention task in patients with schizophrenia: electrophysiological approach Marc Hoonakker, Nadège Doignon-Camus, Elisabeth Bacon, Anne Bonnefond INSERM U1114, Pôle de Psychiatrie-Hôpital Civil de Strasbourg, 1 place de l'Hôpital, Strasbourg, France Keywords: schizophrenia, sustained attention, time-on-task, cognitive control Schizophrenia is characterized by clinical symptoms and by cognitive deficits, likely to play an important part in adaptation of these patients in their every-day life. Among them, there are deficits in sustained attention which are associated with a difficulty for these patients to maintain efficiently their cognitive activity on a task. This basic attentional process is fundamental for the efficiency of the overall of cognitive processes, and so for all aim– directed behaviors (Sarter et al, 2001). The activation of cognitive control mechanisms is essential for good sustained attention performance. The question of whether or not patients with schizophrenia have difficulty sustaining attention is of high relevance, but it has not been conclusively answered in over four decades of research. In this perspective, the main objective of our researches is to better understand, in a time on task perspective, the specific functioning of cognitive and neural mechanisms underlying cognitive control involved in a sustained attention task in schizophrenia. Schizophrenic patients and healthy controls participated in this study. We used the sustained attention to response task (SART, Robertson et al, 1997) for 30 minutes. Behavioral measures combined with event related potentials measures have been recorded. For their analysis, the task was divided into three 10-minute periods. Our preliminary results indicate that schizophrenic patients are able to maintain a stable level of performance over the course of the task. They also revealed specificities in schizophrenic patients in the recruitment of cognitive control mechanisms which could explain these differences in sustained attention performance.

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P15 - Manipulation of mitochondrial gene expression in plant cells through the import of cytoplasmic male sterility–specific RNAs. Rana Khalid Iqbal, Kevin Magne, Frédérique Weber-Lotfi, José M. Gualberto, and André Dietrich Institut de Biologie Moléculaire des Plantes, CNRS et Université de Strasbourg, 12 rue du Général Zimmer, 67084 Strasbourg, France Keywords: cytoplasmic male sterility, mitochondria, plant, RNA trafficking Rearrangements affecting the mitochondrial genome in higher plants often lead to cytoplasmic male sterility (CMS), a trait of particular relevance for breeders. We seek to better understand these mechanisms and to find whether it is conceivable to generate directed CMS. In the absence of methodologies to transform mitochondria in plant cells, our laboratory developed an RNA trafficking strategy to manipulate the organellar genetic system. Plant mitochondria import one third to one half of their tRNAs from the cytosol, including tRNAs valine. The laboratory uses a viral sequence that mimics tRNA valine as a shuttle to import 5’-end attached sequences of interest into mitochondria. The shuttle, called PKTLS, consists of the last 120 nucleotides at the 3’-end of the Turnip yellow mosaic virus genomic RNA. In maize, type S CMS is associated with the presence of specific mitochondrial transcripts carrying two open reading frames, orf355 and orf77. The orf77 contains three regions that are almost identical to regions of the atp9 gene. Due to this similarity, orf77 is considered to be the main cause of CMS-S in maize. In such a context, we express the orf77 RNA from a nuclear transgene in combination with the PKTLS mitochondrial shuttle RNA, so as to target this CMS sequence into the mitochondria of non-CMS Arabidopsis thaliana plants. The transgene is under the control of an inducible promoter. We analyze the impact of mitochondrial targeting of the orf77 RNA on the transcriptome, the organellar functions, the phenotype and the fertility of transformed plants.

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P16 - High throughput screening for selection of anti-HIV drugs Lesia Kovalenko1,2, Nicolas Humbert1, Mattia Mori3, Steven Harper4, Francois Debaene5, Sarah Sanglier-Cianferani5, Olga Zaporozhets2, Maurizio Botta3, Vincenzo Summa4, Yves Mély1

1Laboratoire de Biophotonique et Pharmacologie, UMR 7213 CNRS, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France 2Analytical Chemistry Department, Taras Shevchenko National University of Kyiv, 64 Volodymyrska street, 01601 Kyiv, Ukraine 3Universita degli Studi di Siena, Dipartimento di Biotechnologie, Chimica e Farmacia, via Aldo Moro 2, I-53019 Siena, Italy 4IRBM Science Park, Department of Chemistry, via Pontina KM 30600, I-00040 Pomezia RM, Italy 5Laboratoire de Spectrométrie de Masse Bio-Organique, IPHC-DSA, UPL, UMR 7178 CNRS, ECPM 25 rue Becquerel, 67087 Strasbourg, France Keywords: HIV, Nucleocapsid protein, inhibitor, high throughput screening The efficacy of the highly active anti-retroviral therapy (HAART) that is currently used for HIV-1 treatment is limited by HIV-1 mutations occurring in response to pharmacological pressure. Due to its key role in the viral life cycle and its highly conserved zinc fingers, the nucleocapsid protein (NC) is a profitable target for overcoming drug resistance. Currently developed NC inhibitors suffer from toxicity and limited specificity. Since the specific nucleic acid recognition and chaperone properties of NC are mediated through its hydrophobic platform at the top of the folded zinc fingers, this platform appears as an ideal target for new antiviral strategies with greater specificity. Here we combined molecular modeling, organic synthesis and biophysical studies to find an efficient NC inhibitor. We examined 400 compounds that were previously selected by virtual screening for their ability to inhibit NC chaperone properties based on a NC-promoted cTAR DNA destabilization test. From the screening, 32 compounds were selected for IC50 titrations, based on their ability to induce more than 25% of inhibition at 100 µM concentration in the screening test. The IC50 values of these compounds were found to be in the range 8 – 900 µM. For the most active compounds, we performed additional mechanism-of-action studies. Taken together our results represent a good starting point for further development of anti-NC HIV drugs.

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P 17 - Intracellular distribution of HIV-1 nucleocapsid protein Lysova Iryna a, Halina Anton 1, Julien Godet 1, Pascal Didier 1, Frederic Przybilla

1, Coralie Spiegelhalter 2, Yves Mely 1

1Laboratoire de Biophotonique et Pharmacologie, UMR 7213 du CNRS, Faculté de Pharmacie, Université de Strasbourg, 74, Route du Rhin, 67401 Illkirch Cedex, France 2Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), INSERM U964 CNRS UMR 7104, Université de Strasbourg, Illkirch, France Keywords: nucleocapsid protein, HIV-1, quantum dots, SPT, STORM, TEM, microscopy The nucleocapsid protein (NCp7) of HIV-1 is a small basic protein containing two zinc fingers which are connected by a short basic peptide linker. Both finger domains implicate in binding zinc with high affinity [1]. NCp7 in the form of mature virion or like domain of the Gag polyprotein plays a key role in the reverse transcription of the viral RNA, integration of the proviral DNA into the host genome, selective packaging of the viral genome RNA, and processing and maturation of the new viral particle [2-3]. Previously it was well shown that NCp7 interact with nucleic acids, but interaction of the NCp7 with host cells components remains largely unexplored. In order to find potential targets for NCp7 in the cell, we introduced quantum dots (semiconductor CdSe nanocrystal) conjugated to NCp7 molecules inside the cells by electroporation. By single particle tracking (SPT) technique, we monitored the movements of the QD-NCp7 inside the cell. According to the results of the SPT QD-NCp7 particles exhibited active direct movement. Furthermore, using high resolution STORM and TEM microscopy techniques, we evidenced that NCp7 likely binds to actin filaments and cellular RNAs. 1. Mely Y et al., Zinc binding to the HIV-1 nucleocapsid protein: a thermodynamic investigation by fluorescence spectroscopy Biochemestry. 1996. 35(16).p5175-5182 2. Thomas JA and Gorelick RJ Nucleocapsid protein function in early infection processes Virus Res. 2008. 134(1-2). P39-63 3. Darlix et al., Flexible nature and specific functions of the HIV-1 nucleocapsid protein J. Mol Biol. 2011. 410(4). p65-581

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P18 - Unravelling the molecular mechanisms that trigger Corticospinal Motor Neurons dysfunction and death in Amyotrophic Lateral Sclerosis Christine Marques1 and Caroline Rouaux1 1INSERM U1118, Mécanismes Centraux et Périphériques de la Neurodégénérescence, Université de Strasbourg, Faculté de Médecine, 11 rue Humann, 67 000 Strasbourg, France Keywords: Amyotrophic Lateral Sclerosis; Cerebral Cortex; Corticospinal Motor Neurons; Transcriptomics. Amyotrophic Lateral Sclerosis (ALS) is the most common neurodegenerative disease of the motor system. It leads to progressive muscle paralysis and death within 5 years, and remains incurable. At the cellular level, ALS is characterized by the combined degeneration of two neuronal populations: the corticospinal motor neurons (CSMN), and the spinal motor neurons (SMN). Despite this precise clinical description, the role of CSMN and more broadly of the cerebral cortex, has never been directly interrogated so far. My PhD project aims at better characterizing the cortical pathology in ALS, and at deciphering the molecular mechanisms that selectively trigger CSMN degeneration during the disease. My hypothesis is that CSMN dysfunction and degeneration centrally contribute to ALS onset and progression. In order to characterize the cortical pathology in ALS, I have performed molecular and histological analyses in wild type and SOD1G86R mice, a rodent model of ALS. My preliminary data show that CSMN progressively degenerate in SOD1G86R mice. Interestingly, in drastic contrast to the well-described spinal pathology, CSMN degeneration takes place in absence of major glial pathology. In addition, I have combined retrograde labeling from the spinal cord and FACS to obtain pure populations of CSMN from wild type and SOD1G86R mice. RNAseq analyses of these pure CSMN populations, collected at different stages of the disease, will enable us to unravel the molecular mechanisms behind CSMN degeneration in ALS. On the long run, this project is intended to inform the development of alternative therapeutic strategies for ALS and related CSMN-specific neurodegenerative diseases.

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P19 - Systemic alteration of polyunsaturated fatty acid composition of lipids in a mouse model of amyotrophic lateral sclerosis Laura Robelin1 and José Luis Gonzalez De Aguilar1

1INSERM, Mécanismes Centraux et Périphériques de la Neurodégénérescence, Université de Strasbourg, UMR_S 1118, Faculté de Médecine, 11 rue Humann, 67085 Strasbourg, France Keywords: Amyotrophic Lateral Sclerosis, Motor Neuron Disease, Polyunsaturated Fatty Acid, Lipid Metabolism Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset disease characterized by degeneration of upper and lower motor neurons, progressive muscle wasting, and paralysis. It is also associated with abnormal regulation of energy homeostasis and dyslipidemia. Here we studied polyunsaturated fatty acid (PUFA) composition of lipids as a part of the dysfunction of lipid metabolism affecting ALS. To this end, we used transgenic mice overexpressing a mutated form of Cu/Zn superoxide dismutase as that found in some ALS patients. Spinal cord, skeletal muscle, liver and blood (serum and cell pellet) were obtained from pre-symptomatic and paralyzed mice. We determined fatty acid composition by gas chromatography, and evaluated expression of genes involved in PUFA biosynthesis by quantitative RT-PCR. Gas chromatography revealed that main changes occurred in liver, serum and blood cell pellet of symptomatic mice, whereas no major changes were detected in spinal cord and muscle. In particular, both ω3/ω6 PUFA ratio and proportion of docosahexaenoic acid (DHA), an increase of which is considered to provide beneficial effects, were higher in symptomatic mice. These changes were accompanied by alterations in gene expression and relative activity of enzymes involved in PUFA biosynthesis. Preliminary data obtained from analysis of patient serum samples corroborated the increase in ω3/ω6 ratio and DHA. Taken together, these findings strongly suggest the presence of peripheral alterations of PUFA metabolism that coexist with motor neuron degeneration. Understanding the influence of these metabolic alterations on the neurodegenerative process will open up new therapeutic perspectives.

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P 20 - Genetics and pathophysiologic mechanisms of tubular aggregates myopathies Georges Arielle PECHE, Johann BÖHM, Catherine KOCH, Jocelyn LAPORTE Equipe Laporte, Département Médecine translationnelle et neurogénétique, CNRS UMR 7104 –INSERM U964, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC)

Congenital myopathies have an estimated prevalence of 1:25 000, affect children as well as adults in all populations. They are potentially lethal and encompass more than 10 etiologies varying in symptoms, complications and treatment options. They can be distinguished by the predominance of particular histological signs on muscle biopsies. Tubular aggregate myopathies (TAM) are progressive muscle disorders with an autosomal recessive or dominant inheritance. They are associated with muscle pain and weakness and typically display abnormal accumulations of membrane tubules in muscle fibers. Our team recently identified STIM1 as the first gene implicated in TAM. STIM1 is a major regulator of calcium homeostasis and it was demonstrated that the identified mutations strongly impact on the calcium level in TAM myoblasts. The nature, formation and pathogenicity of the tubular aggregates are not defined and the link between STIM1 mutations and muscle dysfunction is not understood. Moreover, the genetic basis for a majority of TAM families is not known, precluding genetic counseling and improved disease management. This project aims to identify novel implicated genes, to characterize the pathological mechanism underlying the muscle disorder, and to assess rescue strategies in models.

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P 21 - How do temporal based haptic distortions impact the feeling of control in patients with schizophrenia and in patients with bipolar disorder? Patrick Poncelet, Anne Giersch INSERM Unit 1114, Department of Psychiatry, Fédération de Médecine Translationnelle de Strasbourg, University Hospital of Strasbourg, University of Strasbourg, Strasbourg, France Keywords: Feeling of control, schizophrenia, bipolar disorders, haptic Temporal processing impairments have recently been described in patients with schizophrenia. We explore the hypothesis this makes patients vulnerable during motor action and impact their ability to feel in control. We focus on this question due to a possible impact on delusions of control, i.e. the attribution of one’s own action to an external agent. We designed a new original paradigm based on pointing tasks using a robotic device that can record movement trajectory and create virtual surfaces by generating haptic feedbacks. Participants had to perform vertical pointings on a virtual surface without any visual feedback. In the first session, subjects realized a sequence of pointing actions during which the haptic feedback was slightly postponed in time by 15 ms subliminal distortions or 65 ms supraliminal distortions. This allowed us to evaluate the adaptation of the trajectory to the distortion. In the second session, subjects executed sequences of pointings where we introduced varying numbers of supraliminal or subliminal distortions. Subjects were asked to rate their subjective feeling of control after the sequence. The results indicate that patients with schizophrenia and controls adapt their movement efficiently following a supraliminal distortion. The feeling of control is altered in all groups following supraliminal temporal distortions. However, for subliminal distortions, the feeling in control is altered only in patients with schizophrenia. These results are consistent with the hypothesis that abnormal sensitivity to short asynchronies in patients with schizophrenia could represent a vulnerability factor for the development of delusions of control.

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P22 - Effect of neuropeptides on the maturation of human dendritic cells Quentin Muller, Astrid Hoste, Olivier Chaloin, Evelyne Schaeffer, Christopher Mueller, Vincent Flacher Institut de Biologie Moléculaire et Cellulaire, CNRS, Université de Strasbourg, Institut de Biologie Moléculaire et Cellulaire, 15 Rue René Descartes, 67 084 Strasbourg Cedex, France Keywords: Immunology, Neurosciences, Inflammation, Neuropeptides, C-fibers, Dendritic Cells The skin hosts a dense network of sensory neurons, which react to stimuli such as touch, temperature or tissue injury. In particular, C-type nerve fibers release neuropeptides into the skin upon stimulation. In addition to decreasing the activation threshold of bystander neurons, neuropeptides have been suggested to regulate cutaneous inflammation. However, identifying their target cells and resolving the outcome of this immunomodulation remains challenging in a complex system such as the skin. Cutaneous immune responses are initiated by dendritic cells (DCs). Skin DCs capture antigens in their surroundings and are equipped with danger signal receptors. Upon triggering of these receptors by pathogen-derived molecules or pro-inflammatory mediators, they undergo maturation, a phenotypic change that promotes expression of T-cell stimulation factors and migration towards skin-draining lymph nodes. There, DCs stimulate T cells with antigenic peptides and direct them to the skin, generating antigen-specific immune response. Interestingly, DCs are located near sensory neurons in the skin and exhibit receptors for neuropeptides, suggesting that their maturation could result from indirect sensing of danger via C-type nerve fibers. To test this hypothesis, we are investigating the recognition of purified neuropeptides as danger signals in monocyte-derived DCs and in DCs and other immune cells freshly isolated from human skin biopsies. A better understanding of interactions between sensory neurons and DCs could help us resolve the pathogenesis of autoimmune skin diseases such as psoriasis and the regulation of immune system by sensory neurons during inflammation.

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P23 - The CD1-Valproate mouse: a model for studying autism. Sébastien Roux, Jean-Louis Bossu CNRS UPR3212, INCI, Strasbourg, France Keywords: Autism, Sodium Valproate, CD1 Mice, Cerebellum, Purkinje Cells Autism is a neurodevelopmental pathology with multifactorial origins. Some of them are genetic while others are linked to an abnormal exposition to environmental factors during pregnancy. Post-mortem studies have shown anatomical abnormalities on autistic brains especially into cerebellum, a brain structure known for its role in motor coordination, motor learning and (more recently) cognitive tasks.

Sodium valproate is a major anti-epileptic agent, known to induce autism in child born from epileptic mothers under treatments. Indeed, sodium-valproated injected to pregnant mice can mimic autistic symptoms and create murine models of autism.

We have shown that injecting sodium-valproate to pregnant CD1 mice affects the behavior of CD1 pups. They present an altered motor coordination and impairments in social interactions. Moreover, those afflictions are linked with neuroanatomical changes in cerebellum. We have noticed a decreased number of Purkinje cells, the principal cells in cerebellum in a compartmentalized way. Thus, our results suggest that sodium valproate might affect normal neurodevelopment of cerebellum in CD1 mice causing both behavioral and anatomical changes, the main characteristics of autism.

   

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P24 - Physiopathology and preclinical approaches in centronuclear myopathies Tasfaout H, Cowling B, Buono S, Kretz C, Laporte J Department of Translational Medicine and Neurogenetics, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Illkirch, France ; Inserm, U964, Illkirch, France ; CNRS, UMR7104, Illkirch, France Keywords: centronuclear myopathies, Dynamin2, Myotubularin1, BIN1, gene therapy Centronuclear myopathies (CNM) are a group of rare severe muscle diseases characterized by muscle weakness. Muscle biopsies from CNM patients show hypotrophic fibers with centralized nuclei. Three forms have been described: X-linked, Autosomal-recessive (AR) and Autosomal-dominant forms caused by mutations in Myotubularin (MTM1), Amphiphysin 2 (BIN1) and Dynamin 2 (DNM2) respectively. However, the physiopathological mechanisms are barely understood and no specific therapy is available. Using a novel “cross therapy” approach, our group has shown that genetic reduction of Dnm2 in the Mtm1 Knock-out (KO) mice restores a normal lifespan (from 1-3 months to 2 years) with improved muscle structure and function. Therefore, we aim to translate this proof-of-principal experiment by reducing Dnm2 expression using deliverable agents. 14 shRNA sequences and Antisense Oligonucleotides that target specifically mRNA and pre-mRNA respectively were selected and screened on human embryonic Kidney (HEK) and/or mouse C2C12 myoblasts. The best candidates were then injected into wild-type and Mtm1 KO mice to confirm the reduction of Dnm2 detected in cells. The overall goal is to understand the physiopathology of CNM, and to test different novel therapeutic approaches in preclinical trials.

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P25 - The expression of tumor suppressor PTPRD is down-regulated in the liver of patients with HCV infection and in tumor lesions of patients with hepatocellular carcinoma Nicolaas Van Renne1,2, Francois H.T. Duong3, Claire Gondeau4, Diego Calabrese3, Nelly Fontaine1,2, Amina Ababsa1,2, Sarah C. Durand1,2, Markus H. Heim3, Thomas F. Baumert1,2,5,6, Joachim Lupberger1,2

1Inserm U1110, Institut de Recherche sur les Maladies Virales et Hépatiques, Strasbourg 2Université de Strasbourg 3University of Basel, Department of Biomedicine, Hepatology Laboratory, Basel, Switzerland 4Inserm U1040, Biotherapy Research Institute, Montpellier 5Institut Hospitalo-Universitaire, Strasbourg 6Pôle Hépato-digestif, Nouvel Hôpital Civil, Strasbourg Chronic hepatitis C virus (HCV) infection is a main cause of chronic liver disease including hepatocellular carcinoma (HCC). HCV contributes to HCC development directly by viral proteins and indirectly by signal transduction. Signaling is tightly regulated by protein phosphatases and their aberrant expression is involved in various diseases and syndromes. However, the global impact of HCV infection on the expression of protein phosphatases and their impact on liver disease are unknown. To identify the impact of HCV on protein phosphatases we studied the expression of 84 pathway or disease-focused phosphatases in liver biopsies of HCV-infected patients. Using real-time PCR we identified 29 phosphatases that are significantly (p<0.01, Mann-Whitney U-test) up- or downregulated. Among those, several were associated with cancerogenesis including tumor suppressor gene phospho-tyrosine phosphatase, receptor type D (PTPRD). Downregulation of PTPRD expression in HCV-infected liver tissue was validated by fluorescence in situ hybridization (FISH) and Western blotting, while PTPRD expression is not impaired in HBV-infected liver tissue. Moreover, PTPRD expression was downregulated in primary human hepatocytes infected with HCVcc (strain JFH1), while IFN-alpha treatment had no impact on PTPRD. Taken together, it suggests that HCV specifically and directly impairs PTPRD expression in hepatocytes independent from the induced innate immune response. We demonstrated that PTPRD expression is impaired in transformed liver cell lines and in tumor lesions of HCC patients, which we confirmed by in silico analysis of a microarray database (Hoshida et al. N. Engl. J. Med. 2008). These data demonstrate that HCC is associated with impaired PTPRD expression.

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P26 - Improvement of Jimpy oligodendrocyte survival by allopregnanolone Wilding AS., Kemmel V., Taleb O., Patte-Mensah C., Mensah-Nyagan AG Biopathologie de la Myéline, Neuroprotection et Stratégies Thérapeutiques, INSERM U1119, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Bâtiment 3 de la Faculté de Médecine, 11 rue Humann, 67 000 Strasbourg, France. Keywords: Glial cell, Leukodystrophy, Myelin, Neuroprotection, Neurosteroid The mouse mutant jimpy is a well known model for human diseases in which the development of myelin is abnormal. Jimpy mutant mice exhibit severe hypomyelination in the central nervous system (CNS) as patients affected by hopomyelinating leukodystrophies, including the Pelizaeus-Merzbacher disease (PMD), that leads to coordination, motor and intellectual function deterioration. PMD is caused by a X-linked mutation affecting major myelin proteolipid proteins (PLP/DM20). The severe hypomyelination in jimpy CNS results from premature oligodendrocyte (OL) death and the inability of surviving OL to produce compact myelin. Therefore, we made the hypothesis that bioactive substances which may be able to increase the survival of jimpy OL cultures may prevent their premature death and improve their capacity to generate compact myelin in vivo. As a first step towards the validation of our hypothesis, here we investigated the effects of allopregnanolone, a neurosteroid exerting neuroprotective and regenerative actions in various models, on the survival of jimpy OL and control cell cultures. In particular, we used the MTT reduction assays to assess the dose and time-dependent effects of allopregnanolone on the viability or survival of jimpy OL cell line mutated for PLP/DM20 (158JP). The wild-type or normal murine OL cell line (158N) cultures were used as controls. Our results showed that allopregnanolone doses ranging from 100 to 500nM significantly increased 158JP cell survival after 1 or 3 h treatment. Interestingly, we observed that allopregnanolone treatments (100-500nM at 1 or 3h) beneficial for 158JP cell cultures did not affect the viability of control 158N cells, the survival of which was optimal but remained similar in the absence or presence of allopregnanolone. The data suggest that allopregnanolone may ameliorate the survival of jimpy OL through the activation of a mechanism repairing selectively intracellular abnormalities evoked by PLP/DM20 mutation. Future investigations will help to identify this mechanism and to determine whether allopregnanolone may also stimulate compact myelin production in surviving jimpy OL. Acknowledgments W.AS. is a PhD fellow supported by the Consortium NeuroRhine (Neurogenesis and Neuroprotection), INTERREG IV Program (EU, FEDER) and the Région Alsace, France. Authors want to thank Marchand C and Stutz C for technical assistance and Dr Ghandour S for providing the initial vials of 158JP and 158N cells.

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