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BACKGROUND Huntington's disease (HD) is a neurodegenerative disorder caused by the expansion of the trinucleotide CAG in the HD gene. While the presence of nuclear aggregates of mutant huntingtin (mHtt) in neurons is a hallmark of HD, the reason behind its toxicity remains elusive. OBJECTIVE The present study was conducted to assess a correlation between the number of mHtt aggregates and the severity of HD symptoms in R6/1 mice. METHODS We investigated correlations between behavioral deficits and the level of nuclear mHtt aggregates in different neuroanatomical regions in 3-month-old R6/1 mice, the age at which a large variability of symptom severity between animals has been observed. RESULTS R6/1 mice were deficient in instinctive and anxiety related behaviors as well as long-term memory capabilities. Significant differences were also found between the sexes; female transgenic mice displayed less severe deficits than males. While the level of mHtt aggregates was correlated with the severity of HD phenotypes in most regions of interest, an opposite relationship also was found for some other regions examined. CONCLUSIONS The obtained results suggest harmful and region-specific roles of mHtt aggregates in HD symptoms.

Laboratory animals are subjected to multiple manipulations by scientists or animal care providers. The stress this causes can have profound effects on animal well-being and can also be a confounding factor for experimental variables such as anxiety measures. Over the years, handling techniques that minimize handling-related stress have been developed with a particular focus on rats, and little attention to mice. However, it has been shown that mice can be habituated to manipulations using handling techniques. Habituating mice to handling reduces stress, facilitates routine handling, improves animal wellbeing, decreases data variability, and improves experimental reliability. Despite beneficial effects of handling, the tail-pick up approach, which is particularly stressful, is still widely used. This paper provides a detailed description and demonstration of a newly developed mouse-handling technique intended to minimize the stress experienced by the animal during human interaction. This manual technique is performed over 3 days (3D-handling technique) and focuses on the animal's capacity to habituate to the experimenter. This study also shows the effect of previously established tunnel handling techniques (using a polycarbonate tunnel) and the tail-pick up technique. Specifically studied are their effects on anxiety-like behaviors, using behavioral tests (Elevated-Plus Maze and Novelty Suppressed Feeding), voluntary interaction with experimenters and physiological measurement (corticosterone levels). The 3D-handling technique and the tunnel handling technique reduced anxiety-like phenotypes. In the first experiment, using 6-month-old male mice, the 3D-handling technique significantly improved experimenter interaction. In the second experiment, using 2.5-month-old female, it reduced corticosterone levels. As such, the 3D-handling is a useful approach in scenarios where interaction with the experimenter is required or preferred, or where tunnel handling may not be possible during the experiment.

International audience; The G9a/G9a-like protein (GLP) histone lysine dimethyltransferase complex and downstream histone H3 lysine 9 dimethylation (H3K9me2) repressive mark have recently emerged as key transcriptional regulators of gene expression programs necessary for long-term memory (LTM) formation in the dorsal hippocampus. However, the role for hippocampal G9a/GLP complex in mediating the consolidation of spatial LTM remains largely unknown. Using a water maze competition task in which both dorsal hippocampus-dependent spatial and striatum-dependent cue navigation strategies are effective to solve the maze, we found that pharmacological inhibition of G9a/GLP activity immediately after learning disrupts long-term consolidation of previously learned spatial information in male mice, hence producing cue bias on the competition test performed 24 h later. Importantly, the inhibition of hippocampal G9a/GLP did not disrupt short-term memory retention. Immunohistochemical analyses revealed increases in global levels of permissive histone H3K9 acetylation in the dorsal hippocampus and dorsal striatum at 1 h post-training, which persisted up to 24 h in the hippocampus. Conversely, H3K9me2 levels were either unchanged in the dorsal hippocampus or transiently decreased at 15 min post-training in the dorsal striatum. Finally, the inhibition of G9a/GLP activity further increased global levels of H3K9 acetylation while decreasing H3K9me2 in the hippocampus at 1 h post-training. However, both marks returned to vehicle control levels at 24 h. Together, these findings support the possibility that G9a/GLP in the dorsal hippocampus is required for the transcriptional switch from short-term to long-term spatial memory formation

This study investigated whether sst2 gene deletion interacts with age and chronic stress exposure to produce exacerbated emotional and cognitive ageing. Middle-aged (10–12 month) sst2 knockout (sst2KO) and wild-type (WT) mice underwent an unpredictable chronic mild stress (UCMS) procedure for 6 weeks or no stress for control groups. This was followed by a battery of tests to assess emotional and cognitive functions and neuroendocrine status (CORT level). A re-evaluation was performed 6 months later (i.e. with 18-month-old mice). UCMS reproduced neuroendocrine and behavioral features of stress-related disorders such as elevated circulating CORT levels, physical deteriorations, increased anxiety- and depressive-like behaviors and working memory impairments. sst2KO mice displayed behavioral alterations which were similar to stressed WT and exhibited exacerbated changes following UCMS exposure. The evaluations performed in the older mice showed significant long-term effects of UCMS exposure. Old sst2KO mice previously exposed to UCMS exhibited spatial learning and memory accuracy impairments and high levels of anxiety-like behaviors which drastically added to the effects of normal ageing. Spatial abilities and emotionality scores (mean z-scores) measured both at the UCMS outcome and 6 months later were correlated with the initially measured CORT levels in middle-age. The present findings indicate that the deletion of the sst2 receptor gene produces chronic hypercorticosteronemia and exacerbates sensitivity to stressors which over time, have consequences on ageing brain function processes.

Exposure to prolonged, unpredictable stress leads to glucocorticoids-mediated long-lasting neuroendocrine abnormalities associated with emotional and cognitive impairments. Excessive levels of serum glucocorticoids (cortisol in humans, corticosterone in rodents) contribute notably to deficits in working memory (WM), a task which heavily relies on functional interactions between the medial prefrontal cortex (PFC) and the dorsal hippocampus (dHPC). However, it is unknown whether stress-induced increases in plasma corticosterone mirror corticosterone levels in specific brain regions critical for WM. After a 6 week-UCMS exposure, C57BL/6 J male mice exhibited increased anxiety- and depressive-like behaviors when measured one week later and displayed WM impairments timely associated with increased plasma corticosterone response. In chronically stressed mice, basal phosphorylated/activated CREB (pCREB) was markedly increased in the PFC and the CA1 area of the dHPC and WM testing did not elicit any further increase in pCREB in the two regions. Using microdialysis samples from freely-moving mice, we found that WM testing co-occurred with a rapid and sustained increase in corticosterone response in the PFC while there was a late, non-significant rise of corticosterone in the dHPC. The results also show that non-stressed mice injected with corticosterone (2 mg/kg i.p.) before WM testing displayed behavioral and molecular alterations similar to those observed in stressed animals while a pre-WM testing metyrapone injection (35 mg/kg i.p.), a corticosterone synthesis inhibitor, prevented the effects of UCMS exposure. Overall, the abnormal regional increase of corticosterone concentrations mainly in the PFC emerges as a key factor of enduring WM dysfunctions in UCMS-treated animals. Keywords: Stress, Prefrontal cortex, Hippocampus, Glucocorticoids, Working memory

Japanese oysters spatfields were recently discovered in the French Mediterranean Thau lagoon farmed for shellfish. This discovery led to interesting issues concerning research for a highly seashell exploited ecosystem. The analyses of various environmental parameters characterized favorable sites and periods for spat collection. Spat collection in Thau lagoon presented high temporal and spatial heterogeneities related to hydrodynamic, predation, competition and trophic resource variability. Here, the results highlight various explicative factors of the Japanese oysters recruitment within Thau lagoon and underline the high ecological variability in the studied system under oligotrophication.

This study investigated whether sst2 gene deletion interacts with age and chronic stress exposure to produce exacerbated emotional and cognitive ageing. Middle-aged (10-12 month) sst2 knockout (sst2KO) and wild-type (WT) mice underwent an unpredictable chronic mild stress (UCMS) procedure for 6 weeks or no stress for control groups. This was followed by a battery of tests to assess emotional and cognitive functions and neuroendocrine status (CORT level). A re-evaluation was performed 6 months later (i.e. with 18-month-old mice). UCMS reproduced neuroendocrine and behavioral features of stress-related disorders such as elevated circulating CORT levels, physical deteriorations, increased anxiety- and depressive-like behaviors and working memory impairments. sst2KO mice displayed behavioral alterations which were similar to stressed WT and exhibited exacerbated changes following UCMS exposure. The evaluations performed in the older mice showed significant long-term effects of UCMS exposure. Old sst2KO mice previously exposed to UCMS exhibited spatial learning and memory accuracy impairments and high levels of anxiety-like behaviors which drastically added to the effects of normal ageing. Spatial abilities and emotionality scores (mean z-scores) measured both at the UCMS outcome and 6 months later were correlated with the initially measured CORT levels in middle-age. The present findings indicate that the deletion of the sst2 receptor gene produces chronic hypercorticosteronemia and exacerbates sensitivity to stressors which over time, have consequences on ageing brain function processes.

Chromatin modifications, especially histone acetylation, are critically involved in gene regulation required for long-term memory processes. Increasing histone acetylation via administration of histone deacetylase inhibitors before or after a learning experience enhances memory consolidation for hippocampus-dependent tasks and rescues age-related memory impairments. Whether acutely and locally enhancing histone acetylation during early consolidation processes can operate as a switch between multiple memory systems is less clear. This study examined the short- and long-term behavioral consequences of acute intra-CA1 administration of the histone deacetylase inhibitor Trichostatin A (TSA) on cue versus place learning strategy selection after a cue-guided water maze task and competition testing performed 1 or 24 h later in mice. Here, we show that intra-CA1 TSA infusion administrated immediately post-training biased young mice away from striatum-dependent cue strategy toward hippocampus-dependent place strategy under training condition that normally promotes cue strategy in vehicle controls. However, concomitant infusions of TSA with either PKA inhibitor, Rp-cAMPS, into CA1 or cAMP analog, 8Br-cAMP, into dorsal striatum failed to bias young mice to place strategy use. Behavioral and immunohistochemical analyses further indicated that post-training TSA infusion in aged mice rescued aging-associated deregulation of H4 acetylation in the CA1 but failed to reverse phosphorylated CREB deficits and to produce strategy bias on the 24 h probe test. These findings suggest that post-training intra-CA1 TSA infusion promotes dynamic shift from striatum toward the hippocampal system in young but not aged animals, and support the possibility of a role for CREB in the TSA-mediated switch between these two memory systems.

International audience; Altered brain somatostatin functions recently appeared as key elements for the pathogenesis of stress-related neuropsychiatric disorders. The hippocampus exerts an inhibitory feedback on stress but the mechanisms involved remain unclear. We investigated herein the role of hippocampal somatostatin receptor subtypes in both stress response and behavioral emotionality using C57BL/6, wild type and sst2 or sst4 knockout mice. Inhibitory effects of hippocampal infusions of somatostatin agonists on stress-induced hypothalamo-pituitary-adrenal axis (HPA) activity were tested by monitoring peripheral blood and local hippocampus corticosterone levels, the latter by using microdialysis. Anxiolytic and antidepressant-like effects were determined in the elevated-plus maze, open field, forced swimming, and stress-sensitive beam walking tests. Hippocampal injections of somatostatin analogs and sst2 or sst4, but not sst1 or sst3 receptor agonists produced rapid and sustained inhibition of HPA axis. sst2 agonists selectively produced anxiolytic-like behaviors whereas both sst2 and sst4 agonists had antidepressant-like effects. Consistent with these findings, high corticosterone levels and anxiety were found in sst2KO mice and depressive-like behaviors observed in both sst2KO and sst4KO strains. Both hippocampal sst2 and sst4 receptors selectively inhibit stress-induced HPA axis activation but mediate anxiolytic and antidepressive effects through distinct mechanisms. Such results are to be accounted for in development of pathway-specific somatostatin receptor agents in the treatment of hypercortisolism (Cushing's disease) and stress-related neuropsychiatric disorders.

Providing phenotypic information, which is accurate, reliable, repeatable and comparable across countries or laboratories, is critical to gain a better understanding of the relationship between genes and phenotypes. So far, it is indeed extremely difficult to combine different sources of phenotypic data from multiple origins, partly because of the variability in the methods of phenotyping. The phenotyping program of livestock involves the definition of complex phenotypes obtained from data integration at different levels (from molecules to herds), the implementation of the latest technologies to accurately characterize at high speed and low cost, the greatest number of animals in a better characterized environment, and the development and sharing of large databases for data analysis and modeling. Such a program also involves the construction of a coordinated network of research and professional facilities and a common language with shared definition of unambiguous animal traits and of methods to assess them. To this end, it will build on the ‘Animal Trait Ontology of Livestock’ (ATOL) project with the objective of defining precisely the phenotypes of interest for farm animals. Then, it will be necessary to combine an environmental information system related to animal husbandry and associated methods to capture the phenotypic differences between animals.

This study examines the effects of ibotenic acid lesions of the mediodorsal nucleus of the thalamus (MD) on serial contextual memory retrieval in non-stress and stress conditions. Independent groups of mice learned two successive contextual serial discriminations (D1 and D2) in a four-hole board. The discriminations differed each by the color and texture of the floor. Twenty-four hours later, memory testing occurred in independent groups of mice on one of the two floors of the initial acquisition session. Half of the subjects received three electric footschocks (0.9 mA, 2 s) 5 min prior to testing. Results showed that (i) stress induced a plasma corticosterone rise of same magnitude in sham-operated and MD-lesioned mice; (ii) non-stressed sham-operated mice accurately remembered D1 but not D2, whereas stressed sham-operated animals remembered D2 but not D1; (iii) non-stressed MD-lesioned mice exhibited a memory retrieval pattern similar to that observed in non-stressed sham-operated mice; (iv) however, the stress-induced inversion of the memory retrieval pattern was not observed in MD animals. The effects of MD lesions on memory retrieval in this task are similar to those observed in earlier studies in prefrontal cortex or amygdala-lesioned mice [Chauveau F, Pierard C, Coutan M, Drouet I, Liscia P, Beracochea D. Prefrontal cortex or basolateral amygdala lesions blocked the stress-induced inversion of serial memory pattern in mice. Neurobiol Learn Mem 2008;90:395–403]; they are however in sharp contrast with mice exhibiting hippocampal lesions [Chauveau F, Pierard C, Tronche C, Coutan M, Drouet I, Liscia P, et al. The hippocampus and prefrontal cortex are differentially involved in serial memory retrieval in non-stress and stress condition. Neurobiol Learn Mem; in press; Chauveau F, Pierard C, Tronche C, Coutan M, Drouet I, Liscia P, et al. Rapid stress-induced corticosterone rise in the hippocampus reverses serial memory retrieval pattern. Hippocampus; in press]. Overall, the present findings highlight the involvement of the MD in an AMG/PFC system mediating the rapid effects of stress on serial memory retrieval.

APP(751SL) mice of 5-6- and 7-8-month-old and their wild-type littermates were submitted to one-session learning in a water-maze with three levels of training (4, 12 or 22 trials). Training consisted in finding a submerged platform with a fixed location and marked by a cue. During testing two platforms were presented: one consistent with the spatial location allowing place-response (PR) and the other signaled by the cue enabling cued-response (CR). When testing occurred 24h after training, wild-type and 5-6-month-old APP(751SL) mice exhibited a shift in response strategy as a function of training level, by executing CR when trained with 4 trials and PR when trained with 12 trials, but 7-8-month-old APP(751SL) mice executed only CR. However, they displayed PR when tested 1h after 12- and 22-trial, suggesting a consolidation deficit. Zif268 imaging showed plasticity impairment of the hippocampal-dependent memory system but not of the dorsolateral caudate nucleus. Moreover, in these APP(751SL) mice, the deficit selectively affecting hippocampal function cannot be directly related to the onset of beta-amyloid deposits.

Somatostatin (SS14) has been implicated in various cognitive disorders, and converging evidence from animal studies suggests that SS14 neurons differentially regulate hippocampal- and striatal-dependent memory formation. Four SS14 receptor subtypes (SSTR1-4) are expressed in the hippocampus, but their respective roles in memory processes remain to be determined.In the present study, effects of selective SSTR1-4 agonists on memory formation were assessed in a water-maze task which can engage either hippocampus-dependent "place" and/or striatum-dependent "cue" memory formation.Mice received an intrahippocampal injection of one of each of the selective agonists and were then trained to locate an escape platform based on either distal cues (place memory) or a visible proximal cue (cue memory). Retention was tested 24 h later on probe trials aimed at identifying which memory strategy was preferentially retained.Both SS14 and the SSTR4 agonist (L-803,087) dramatically impaired place memory formation in a dose-dependent manner, whereas SSTR1 (L-797,591), SSTR2 (L-779,976), or SSTR3 (L-796,778) agonists did not yield any behavioral effects. However, unlike SS14, the SSTR4 agonist also dose-dependently enhanced cue-based memory formation. This effect was confirmed in another striatal-dependent memory task, the bar-pressing task, where L-803,087 improved memory of the instrumental response, whereas SS14 was once again ineffective.These data suggest that hippocampal SSTR4 are selectively involved in the selection of memory strategies by switching from the use of hippocampus-based multiple associations to the use of simple dorsal striatum-based behavioral responses. Possible neural mechanisms and functional implications are discussed.

Previous studies showed that the optimization of behavioral performance through extended training depends on a switch from hippocampus-based memory to striatum-based habit. Here we investigate whether the amount of training within one learning session influences the retention of memory for hippocampal versus striatal strategies. Mice were trained to search for a submerged cue-marked platform which remained in the same spatial location in the water-maze for each of three training regimens (4, 12 or 22 trials). Subsequently, they were either tested for retention of memory 1 h or 24 h later on a probe test or killed at different time points over a 7-h period to determine the kinetic of cAMP response element binding protein (CREB) phosphorylation in both memory systems. During the probe test mice had to choose between a submerged platform located in the same position as during the acquisition phase (spatial solution) and a platform marked by the cue but located in the opposite quadrant of the pool (cue-guided solution). Results showed that the animals first preferred the cue-marked platform, which represents a strategy that was selectively impaired by lesions of the dorsolateral caudate-putamen. With further practice, or context pre-exposure, animals transiently favored the hippocampus-dependent place solution but finally, both strategies became interchangeable and insensitive to either lesion. CREB phosphorylation increased in both memory systems following acquisition but training-dependent changes selectively occurred in the hippocampus wherein biphasic activation was initiated by the four-trial training and blocked by training for 22 trials. These findings indicate that learning in one session consists of three acquisition stages with parallel engagement of multiple memory systems at the beginning of learning. They suggest, however, that, in a later phase, dynamic interplays promote the use of the most adapted brain system depending on practice and this is accompanied by specific patterns of CREB phosphorylation in the hippocampus.

Several lines of evidence indicate that Ca2+/calmodulin-stimulated isoforms of adenylyl cyclase (AC) are involved in long-term potentiation and in certain forms of learning. Recently, we found that training in different types of learning task differentially activates Ca2+-sensitive versus Ca2+-insensitive AC activities in certain brain regions, indicating that AC species other than those stimulated by Ca2+/calmodulin may play an important role in learning processes ( Guillou, Rose, & Cooper, 1999 ). Here, we report the effects of spatial reference memory training in a radial arm maze on the levels of AC1 and AC2 mRNA in the dorsal hippocampus of C57BL/6 mice. Acquisition of the task was associated with a learning-specific and time-dependent increase of AC1 mRNA expression selectively in subfields CA1–CA2. In contrast, AC2 mRNA levels were either reduced or not reliably affected depending on the stage of acquisition. Moreover, no significant changes in AC expression were observed either in the dorsal hippocampus of mice trained in a non-spatial (procedural) version of the task or in cortical regions of mice learning the spatial or procedural task. The regional specificity of these effects indicates that the formation of spatial and non-spatial memory requires distinct contributions from Ca2+-sensitive and Ca2+-insensitive AC in the hippocampus. It is suggested that downregulation of AC2 throughout all hippocampal subfields may play a permissive role during the acquisition of spatial learning whereas an upregulation of AC1 specifically in subfield CA1, may be critical to accurately encode, store or use spatial information.

Previous studies showed that injections of somatostatin (SS-14) into the hippocampus facilitate the acquisition of spatial tasks in mice. The present study was aimed at better understanding the learning and memory processes that could be affected by hippocampal SS-14 stimulation. Balb/c mice were submitted to a two-stage learning paradigm. In stage 1, they were trained for acquisition of a spatial discrimination task in a radial maze and, in stage 2, were submitted to a probe test aiming at evaluating their ability to use flexibly their previously acquired knowledge. Injections of vehicle or SS-14 were given during the acquisition phase and/or before the probe test using a 2 x 2 factorial design. Mice receiving SS-14 during acquisition failed to succeed in the probe test despite showing a trend to faster acquisition of the initial spatial discrimination task. By contrast, when given only prior to probe trials, SS-14 did not yield any behavioural effects. Thus, SS-14 interfered with the establishment of a flexible form of memory, not with its expression per se, and therefore dissociated the acquisition of place responses from their flexible use. The theoretical issues raised by the present findings are discussed.

In an attempt to characterize the contribution of the 5-HT1B receptor to behavior, 5-HT1B knock-out (KO) mice were subjected to a battery of behavioral paradigms aimed at differentiating various components of cognitive and emotional behaviors. In an object exploration task, wild-type (WT) and 5-HT1B KO mice did not differ in locomotor activity. 5-HT1B KO mice, however, displayed lower thigmotaxis (an index of anxiety) associated with a higher level of object exploratory activity, but no genotype differences were observed in the elevated plus maze. 5-HT1B KO mice also displayed a lack of exploratory habituation. In the spatial version of the Morris water maze, 5-HT1B KO mice showed higher performances in acquisition and transfer test, which was not observed in the visual version of the task. No genotype differences were found in contextual fear conditioning, because both WT and 5-HT1B KO mice were able to remember the context where they had received the aversive stimulus. The deletion of the 5-HT1B receptor, associated with appropriate behavioral paradigms, thus allowed us to dissociate anxiety from response to novelty, and perseverative behavior (lack of habituation) from adaptive behavioral inhibition underlying cognitive flexibility (transfer stage in the water maze). The deletion of the 5-HT1B receptor did not result in significant developmental plasticities for other major 5-HT receptor types but may have influenced other neurotransmission systems. The 5-HT1B receptor may be a key target for serotonin in the modulation of cognitive behavior, particularly in situations involving a high cognitive demand.

La qualité sensorielle de la viande bovine est un critère important d’appréciation par lesconsommateurs qui demeurent insatisfaits en particulier de la tendreté. Celle-ci a une origine complexeet multifactorielle, qui la rend difficilement maîtrisable par la filière bovine. À l’issue de plusieurs annéesde recherche, notre équipe a établi une liste de marqueurs de la tendreté de la viande. Dans cetteliste, les Heat Shock Proteins (Hsp) figurent à des « carrefours » biologiques dans l’interactomemusculaire lié à la tendreté. Parmi elles, la Hsp27 est différentiellement exprimée entre des musclesde tendreté extrême. Comprendre le rôle de Hsp27 et de ses interacteurs dans l’établissement de latendreté est un des enjeux des recherches en production de viande.Afin d’atteindre cet objectif, l’invalidation du gène hspb1 (codant la protéine Hsp27) a été réalisée parrecombinaison homologue (KO) chez la souris (en collaboration avec l'IGFL et le PBES Lyon). Cemodèle a permis d’analyser les conséquences de l’invalidation de hspb1 sur le développement et lescaractéristiques des tissus liés à la qualité (muscle et tissus adipeux), ainsi que sur la fonctionnalité del’interactome "tendreté". Ce travail s’insérait dans l’Action Incitative du Département Phase« Phénotypage des animaux modèles » (2009-2012).Les souris KO hspb1-/- sont viables et fertiles, ne présentent aucune anomalie apparente mais ont unformat plus petit que celui de leurs témoins (Kammoun et al, 2013a). Au niveau plasmatique, uneinteraction génotype x âge x sexe a été révélée pour 16 paramètres biologiques sur les 36 indicateursmétaboliques mesurés. Les principales différences détectées entre KO et témoins suggèrent desdifférences de métabolismes hépatique et lipidique chez les mâles et de métabolisme du glucose et del’acide urique chez les femelles. Aucune différence n'a été observée pour les paramètresreprésentatifs du métabolisme musculaire. La structure musculaire des animaux a été analysée defaçon complémentaire par microscopie optique et électronique à transmission. La première approche n'a pas permis de révéler de différences dans les caractéristiques des fibres musculaires (typecontractile et métabolique, forme, périmètre, surface de section) à l'exception d'une tendance pour uneproportion plus élevée de fibres de petite taille chez les souris KO. Toutefois, l'analyse des profilsélectrophorétiques des isoformes de chaînes lourdes de myosine suggèrent un retard de maturationmusculaire chez les souris KO. Au niveau ultrastructural, les fibres des animaux KO sont caractérisées,comparativement à celle des témoins, par un appareil contractile musculaire moins organisé (Figure 1)qui devient très déstructuré 72h post-mortem . Ainsi le phénotype musculaire fin des souris KO estaltéré.Une analyse bioinformatique a été réalisée dans l'objectif de compléter la liste des interacteurs de laprotéine Hsp27 et des gènes cibles de l’invalidation d’hspb1 susceptibles de participer à desdifférences de structure du muscle et de la tendreté. Les partenaires ou cibles prédits de Hsp27 sontdes protéines impliquées dans différentes fonctions, comme des Hsps (Hsp20, Cryab, Hsp70a1a etHsp90aa1), des régulateurs de l'apoptose (Fas, Chuk, et caspase-3), des facteurs de traduction(Eif4E et Eif4G1), des protéines du cytosquelette (Desmine) et des antioxydants (Sod1). Lesabondances de 15 protéines ont été quantifiées par Western blot dans deux muscles de souris KO etde leurs témoins. Elles sont modifiées chez les souris dépourvues de Hsp27 principalement dans lemuscle le plus oxydatif (Kammoun et al., 2012, 2013b). Cette étude démontre l'existence de liensfonctionnels entre Hsp27 et ses cibles prédites.L'ensemble des données issues de cette étude réalisée dans une espèce modèle apporte desconnaissances nouvelles sur les mécanismes moléculaires impliqués dans l’établissement de latendreté de la viande bovine. Elle suggère que le statut en Hsp, les processus apoptotiques et laprotection contre le stress oxydatif contribue à l'évolution de l'ultrastructure post-mortem des muscleset à la tendreté de la viande. Ces données seront complétées par les résultats d'une analyseprotéomique en cours. Ces nouvelles connaissances seront validées ultérieurement sur muscle bovin.

The multiple memory systems hypothesis posits that different neural circuits function in parallel and may compete for information processing and storage. For example, instrumental conditioning would depend on the striatum, whereas spatial memory may be mediated by a circuit centered on the hippocampus. However, the nature of the task itself is not sufficient to select durably one system over the other. In this study, we investigated the effects of natural and pharmacological rewards on the selection of a particular memory system during learning. We compared the effects of food- or drug-induced activation of the reward system on cue-guided versus spatial learning using a Y-maze discrimination task. Drug-induced reward severely impaired the acquisition of a spatial discrimination task but spared the cued version of the task. Immunohistochemical analysis of the phosphorylated form of the cAMP response element binding (CREB) protein and c-Fos expression induced by behavioral testing revealed that the spatial deficit was associated with a decrease of both markers within the hippocampus and the prefrontal cortex. In contrast, drug reward potentiated the cued learning-induced CREB phosphorylation within the dorsal striatum. Administration of the protein kinase A inhibitor 8-Bromo-adenosine-3',5'-cyclic monophosphorothioate Rp isomer (Rp-cAMPS) into the dorsal striatum before training completely reversed the drug-induced spatial deficit and restored CREB phosphorylation levels within the hippocampus and the prefrontal cortex. Therefore, drug-induced striatal hyperactivity may underlie the declarative memory deficit reported here. This mechanism could represent an important early step toward the development of addictive behaviors by promoting conditioning to the detriment of more flexible forms of memory.

The involvement of hippocampal somatostatin (SS-14) in spatial discrimination learning in mice was evaluated by bilaterally administering either SS-14 or its depletor, cysteamine, into the hippocampus through chronically implanted guide cannulae. Two separate experiments were performed using an eight-arm radial maze. In Experiment 1, mice were trained for 4 days on a discrimination task in which only three arms (A, B, and C) were used. In each daily session, subjects were each presented in 10 trials with the pair AB and the pair BC, with arm B consistently baited over trials (e.g., A − B + and B + C −). Immediately after completion of the fourth daily session with these regular trials, they were given 10 additional probe trials with free access to all three arms (e.g., A − B + C −). In Experiment 2, animals had free access to all eight arms. They were trained to discriminate the three arms that were consistently baited over trials (8 trials per day for 7 days). Intrahippocampal injections were given each day, 30 min before testing. The results of both experiments showed that, compared with saline, SS-14 administration (0.2 µg/0.2 µl per hippocampus) resulted in faster acquisition rates. In contrast, animals treated with cysteamine (20 µg) were dramatically impaired. In Experiment 1, however, the SS-14 treatment group was significantly impaired relative to both the saline and the intact control groups when testing was shifted from regular to probe trials. The impaired ability of somatostatin-treated mice to adapt to probe trials, together with their improved performance on both types of spatial discrimination task, suggests that this peptide produced a change in the strategy the animals used to solve these tasks. Notwithstanding, the present findings seem difficult to reconcile with current theories about hippocampal function.

In order to better understand environmental disturbances in the French coastal Mediterranean lagoons, we used an ecotoxicological approach based on the measurement of the toxicity of the sediments using oyster embryo bioassay that provides a basis for assessing the effects on the fauna of contaminants adsorbed on the sedimentary particles. The study covers all of the main lagoons of the French Mediterranean coasts of Languedoc Roussillon, Camargue, and Provence (Berre and Bolmon lagoons), where 188 stations were sampled. The toxicity tests provide evidence of variable levels of toxicity in sediments. Contaminated lagoons such as La peyrade, Le canet, and Ingrill and locally affected lagoons such as Bages-Sigean, Vaccares, Bolmon, and Berre have sampling stations with 100% of larval abnormalities during 24-h development. In all of the lagoons, the toxicity was mainly located close to local harbors and rivers. Salses Leucate (Languedoc roussillon) lagoon was found very clean, with no important toxicity. The results are discussed in terms of environmental disturbances of the coastal lagoons and with regard to the long-term monitoring of the impact of contaminants on the coastal environment.

The aim of this study was to determine if the memory of the association between a step-down response and a foot-shock can be dissociated from the memory of the context in which the shocking experience occurred. To test this, two versions of the step-down inhibitory avoidance task were used: a standard version, in which animals were given one trial with a weak exposure to the context and a new version, in which animals were given a stronger exposure to the context. A retention test was performed with the platform placed either in the same conditioning chamber as during the acquisition phase or in a new context. Our results demonstrate that the step-down inhibitory avoidance can actually be solved without a functional hippocampus. Specifically, the results show that hippocampus-lesioned mice and sham controls can express similar level of memory performance but use two different strategies which were distinguished by assessing retention in a new context. Hippocampus-lesioned mice and mice injected with forskolin (adenylyl cyclase activator) 3 h after acquisition use a memory strategy which is independent of the context of acquisition. In addition, our results confirm that the cAMP signaling pathway is a key step in memory consolidation processing.

The hippocampal somatostatin (sst) receptor subtype 4 (sst(4)) modulates memory formation by diminishing hippocampus-based spatial function while enhancing striatum-dependent behaviors. sst(4)-mediated regulations on neuronal activity in the hippocampus appear to depend on both competitive and cooperative interactions with sst receptor subtype 2 (sst(2)). Here, we investigated whether interactions with sst(2) receptors are required for sst(4)-mediated effects on hippocampus-dependent spatial memory and striatum-dependent cued memory in a water maze paradigm. Competition was assessed in mice by intrahippocampal injections of the sst(4) agonist L-803,087 alone or combined with sst(2) agonists (L-779,976 or octreotide). Effects of L-803,087 were also tested in sst(2) knockout mice to assess for receptor cooperation. Finally, sst(2a) and sst(4) localizations within hippocampal subregions were analyzed by immunohistochemistry and expression levels of sst(2a) and sst(2b) were quantified by real-time qPCR. Hippocampal injections of L-803,087 impaired spatial memory but enhanced cued memory. The latter effect was lost not only in sst(2) knockout mice but also in the presence of sst(2) agonists, whereas the former effect remained unaffected by sst(2) agonists or gene deletion. Octreotide and L-779,976 did not yield memory effects but reduced swim velocity throughout the acquisition trials suggesting that stimulation of sst(2) affected motivation and/or anxiety. sst(2a) and sst(4) were respectively detected in the dentate gyrus (DG) and the CA1 subfield suggesting that their functional interactions are not mediated by direct receptor coupling. Hippocampus sst(2a) expression was 36-fold higher than sst(2b). Possible neural mechanisms and functional significances for interaction between memory systems in relationship with stress-induced changes in hippocampal functions are discussed.

According to the “multiple memory systems” hypothesis, different forms of memory are organized in independent brain systems. In this context, it is now widely accepted that “declarative” forms of memory depend on hippocampal circuitry, whereas “habit,” “procedural,” or “response memory” depend upon a dorsal striatal system (Cohen et al. 1997). Numerous data indicate that multiple memory systems are coactivated in parallel during learning and that these systems interact (for review, see White and McDonald 2002). Converging evidence from recent human neuroimaging research and animal studies suggests that the nature of the interactions between hippocampus and dorsal striatal memory systems are either competitive (for review, see Poldrack and Packard 2003; McDonald et al. 2004a) or cooperative (Voermans et al. 2004; for review, see McDonald et al. 2004b). Despite the considerable interest in describing the cellular and molecular bases that underlie memory formation, the potential neuronal mechanisms mediating such interactions remain largely unknown. Studies have nevertheless indicated that levels of glutamate or acetylcholine release in both structures might play a crucial role (Packard 1999; McIntyre et al. 2003). Further, different types of learning have been associated with changes in the activity of particular adenylyl cyclase (AC) subtypes in a mammalian brain (for review, see Mons et al. 1999; Mons and Guillou 2004). Current data support the idea that calcium-insensitive AC isoforms may subserve different forms of memory. For instance, decreased calcium-insensitive AC activity was observed in the hippocampus after spatial learning in a water-maze task, whereas, in contrast, an increase in this enzyme activity was found after acquisition in a procedural version of the task or in a bar-pressing task (Guillou et al. 1998, 1999a). Furthermore, the expression of the calcium-insensitive AC2 in the hippocampus was found to selectively decrease during acquisition of spatial tasks (Mons et al. 2003, 2004). Task-dependent opposite regulations of calcium-insensitive ACs strongly suggest that the particular direction of this regulation of cAMP formation within the hippocampus could reflect a mechanism enabling a selection between competing memory systems. As previously suggested, cAMP signaling deriving from calcium-insensitive ACs could have an inhibitory as opposed to a permissive action on hippocampal functioning (Guillou et al. 1999a). The aims of the present study were to investigate whether stimulation of hippocampal ACs subtypes blocks the formation of hippocampus-dependent memory and facilitates the use of memories mediated by competitive systems and, particularly, to determine at what time period during the consolidation phase that follows initial acquisition this cAMP-mediated mechanism is critical. To investigate this issue, 24-h retention performances of mice were measured following post-training stimulation (0–9 h) of hippocampal ACs by intrahippocampal injections of forskolin (FK) because pharmacological tools selective for particular AC isoforms are not yet available. The injections were given after either a bar-pressing task or water-maze tasks, which challenge either striatal-dependent “response memory” or hippocampus-dependent “place memory” (Packard et al. 1994; for review, see Kelley 2004). In addition, the kinetics of the phosphorylation of the cAMP-responsive element binding (CREB) was examined using an immunohistochemical technique to monitor the impact of the FK injection on cAMP-dependent neuronal activation in the brain.

Ca(2+) regulates mammalian adenylyl cyclases in a type-specific manner. Stimulatory regulation is moderately well understood. By contrast, even the concentration range over which Ca(2+) inhibits adenylyl cyclases AC5 and AC6 is not unambiguously defined; even less so is the mechanism of inhibition. In the present study, we compared the regulation of Ca(2+)-stimulable and Ca(2+)-inhibitable adenylyl cyclases expressed in Sf9 cells with tissues that predominantly express these activities in the mouse brain. Soluble forms of AC5 containing either intact or truncated major cytosolic domains were also examined. All adenylyl cyclases, except AC2 and the soluble forms of AC5, displayed biphasic Ca(2+) responses, suggesting the presence of two Ca(2+) sites of high ( approximately 0.2 microM) and low affinity ( approximately 0.1 mM). With a high affinity, Ca(2+) (i) stimulated AC1 and cerebellar adenylyl cyclases, (ii) inhibited AC6 and striatal adenylyl cyclase, and (iii) was without effect on AC2. With a low affinity, Ca(2+) inhibited all adenylyl cyclases, including AC1, AC2, AC6, and both soluble forms of AC5. The mechanism of both high and low affinity inhibition was revealed to be competition for a stimulatory Mg(2+) site(s). A remarkable selectivity for Ca(2+) was displayed by the high affinity site, with a K(i) value of approximately 0.2 microM, in the face of a 5000-fold excess of Mg(2+). The present results show that high and low affinity inhibition by Ca(2+) can be clearly distinguished and that the inhibition occurs type-specifically in discrete adenylyl cyclases. Distinction between these sites is essential, or quite spurious inferences may be drawn on the nature or location of high affinity binding sites in the Ca(2+)-inhibitable adenylyl cyclases.

Cysteamine was used as a tool aimed at investigating the role of central somatostatin (SS-14) and was shown to modulate learning in a task-dependent manner. However, direct arguments have not yet been provided to support the hypothesis that impairments and facilitation of learning produced by cysteamine are both mediated by the hippocampus. Mice were given daily intrahippocampal injections of artificial cerebrospinal fluid (CSF) or cysteamine at doses of either 2.5 microg/0.2 microl or 25 microg/0.2 microl 1 h prior to each learning session of a bar-pressing task, for which the acquisition was previously shown to be improved by systemic injections. The results showed that, with respect to CSF, the mice injected with cysteamine learned the bar pressing task faster whereas no evidence of changes in locomotor activity was provided. Moreover, the results showed that retention was specifically increased in the two groups injected with cysteamine. It is argued that the action of cysteamine on the hippocampus is sufficient to modulate specifically learning-memory processes in a task-dependent manner. In conclusion, the blockade of some hippocampal information processing function by cysteamine is discussed to understand the bidirectional effects of drugs on learning and memory.

Adenylyl cyclases (ACs) are involved in a variety of advanced CNS functions, including some types of learning and memory. At least nine AC isoforms are expressed in the brain, which are divisible into three broad classes based on the ability of Ca2+to modulate their activity. This study examined the hypothesis that different learning tasks would differentially activate ACs in selected brain regions. The ability of forskolin or Ca2+to enhance AC activity in the hippocampus, parietal cortex, striatum, and cerebellum was examined after mice had been trained in either a spatial or procedural learning task using a Morris water maze. Sensitivity of ACs to forskolin was enhanced to a greater degree in most brain regions after procedural learning, but Ca2+-sensitive ACs in the hippocampus were more sensitive to spatial learning. Because nonspecific behavioral elements, such as stress or motor activity, were similar in both experimental tasks, these results provide the first evidence that acquisition of different kinds of learning is associated with selective changes in particular AC species in a mammalian brain and support the idea that different biochemical processing, involving particular isoforms of ACs, subserves different memory systems.

Evidence from systems as diverse as mollusks, insects and mammals has revealed that adenylyl cyclase, cyclic adenosine 3′,5′-monophosphate (cAMP) cascade, cAMP-dependent protein kinases and their substrates are required for the cellular events underlying the short-term and long-term forms of memory. In Aplysia and Drosophila models, the coincident activation of independent paths converge to produce a synergistic activation of Ca2+/calmodulin-stimulable adenylyl cyclase, thereby enhancing the cAMP level that appears as the primary mediator of downstream events that strengthen enduring memory. In mammals, in which long-term memories require hippocampal function, our understanding of the role of adenylyl cyclases is still fragmentary. Of the differently regulated isoforms present in the hippocampus, the susceptibility of type 1 and type 8 to stimulation by the complex Ca2+/calmodulin and their expression in the hippocampus suggest a role for these two isoforms as a molecular coincidence device for hippocampus-related memory function. Here, we review the key features of Ca2+/calmodulin stimulable adenylyl cyclases, as well as the involvement of cAMP-regulated signaling pathway in the processes of learning and memory.

The hypothesis of a role for hippocampal somatostatin (SS-14) in learning and memory processes was further examined by means of 2 selective learning tasks that were previously shown to be either impaired (spatial discrimination task) or facilitated (barpressing task) by hippocampal lesions. Results showed that subcutaneous injections of cysteamine (160 mg/kg) (a) impaired acquisition of the spatial task while producing an opposite (i.e., facilitative) effect on acquisition of the barpressing task and (b) produced an up regulation of hippocampal adenylyl cyclase (AC) activity, which was antagonized by spatial discrimination training but enhanced by training in the barpressing task. Moreover, opposite task-dependent training-induced changes in hippocampal AC activity was observed in saline-treated mice. These results suggest that bidirectional regulatory mechanisms of hippocampal function involving both SS-14 and ACs may occur as a function of the type of learning.

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Data showed that chronic stress exposure during middle-age induces persistent stress symptoms such as increased anxiety and depression state leading to long-lasting emotional disorders that can extend to cognitive deficits. Changes in epigenetic regulations, such as histones acetylation and methylation, have been identified as a key mechanism through which stress exposures induce emotional and cognitive alterations. In this study, we investigated the long-lasting behavioral impacts of chronic stress exposures and their epigenetic correlates with aging. C57bl/6 mice underwent an unpredictable chronic mild stress procedure (UCMS) for 6 weeks at either 6 month- (adult) or 11-month old (middle-age) while age-matched controls remained undisturbed (non-stressed groups). All mice underwent a battery of behavioral testing when getting the age of 18 months and then, were killed for immunohistochemical detection of the transcriptional repressive histone H3 tri-methylation at lysine 27 (H3Kme3). Trait anxiety in the elevated plus-maze, but not reactive anxiety in the marble burying test, as well as despair in forced-swimming and tail suspension tests were increased in mice that had experienced UCMS at 11 month as compared to controls. Spatial memory in water-maze and working memory were also further impaired. Principal component analysis confirmed a clear-cut separation of the behavioral patterns in these two groups of mice. These behavioral impairments were associated with changes in histone H3K27me3 in CA1 area of the ventral hippocampus, the prefrontal cortex and lateral nucleus of the amygdala. In contrast, mice exposed to UCMS at 6-month old, did not display any persistent deficits nor any epigenetic changes when getting aged. In conclusion, these results indicate that chronic stress stronger impacts on behavioral emotionality and cognitive aging in association with epigenetic regulatory changes in middle-aged adults. Although adulthood has been associated with an increase in stress resistance, a second phase of vulnerability to stress emerges at midlife and interferes with brain aging., Le stress chronique est un phénomène universel qui concerne toutes les strates de la société, quels que soient l’âge, le sexe, le milieu social, les origines culturelles et géographiques. Néanmoins la vulnérabilité est variable selon l’âge d’exposition. Il est aujourd’hui avéré qu’une exposition chronique à un stress ou bien une exposition à un stress aigu intense chez l’enfant, voire même pendant la période périnatale, peut avoir des conséquences dramatiques sur le développement cérébral et peut engendrer un certain nombre de pathologies neuropsychiatriques plus tard au cours de la vie. Néanmoins, une plus forte résistance et une capacité de résilience apparaît à l’âge adulte se caractérisant notamment par une adaptabilité et la possibilité de retourner rapidement à un état initial après une exposition au stress. Toutefois, des travaux récents suggèrent qu’une seconde phase de vulnérabilité apparait en milieu de vie, période charnière durant laquelle certaines structures du système limbique commencent peu à peu à décliner. L’objectif de cette thèse a été ainsi dans un premier temps de mieux caractériser la nature des perturbations somatiques à court terme et des déficits comportementaux à long terme qui apparaissent suite à un stress chronique durant cette période charnière au cours de laquelle s’installe une vulnérabilité accrue aux stress. Pour répondre à cette première problématique, le modèle de stress chronique léger et imprédictible mis au point chez la souris a été utilisé. Les impacts à court terme ont été déterminés en analysant les troubles somatiques et endocriniens, puis ceux à long-terme (chez le sujet âgé), ont été déterminés en analysant les troubles comportementaux anxieux, dépressifs et cognitifs, caractéristiques du syndrome de stress. D’autre part, il s’agissait d’identifier les réorganisations épigénétiques associés au vieillissement et au stress chronique ayant eu lieu en milieu de vie. Nous nous sommes concentrés sur les perturbations de la marque permissive H3K9ac et la marque répressive H3K27me3 en fonction du vieillissement et du stress chronique à long terme. Enfin, une approche pharmacologique a été utilisée pour évaluer l’impact d’un traitement ciblant ces mécanismes épigénétiques sur les déficits comportementaux. Un traitement chronique de 3 semaines à base de butyrate de sodium (NaB) qui est un iHDAC, a été administré aux animaux après la phase de stress chronique, puis les évaluations comportementales ont été réalisé à long terme, chez les sujets âgés.