LSD1 is an environmental stress-sensitive negative modulator of the glutamatergic synapse.

Along with neuronal mechanisms devoted to memory consolidation -including long term potentiation of synaptic strength as prominent electrophysiological correlate, and inherent dendritic spines stabilization as structural counterpart- negative control of memory formation and synaptic plasticity has been described at the molecular and behavioral level. Within this work, we report a role for the epigenetic corepressor Lysine Specific Demethylase 1 (LSD1) as a negative neuroplastic factor whose stress-enhanced activity may participate in coping with adverse experiences. Constitutively increasing LSD1 activity via knocking out its dominant negative splicing isoform neuroLSD1 (neuroLSD1KO mice), we observed extensive structural, functional and behavioral signs of excitatory decay, including disrupted memory consolidation. A similar LSD1 increase, obtained with acute antisense oligonucleotide-mediated neuroLSD1 splicing knock down in primary neuronal cultures, dampens spontaneous glutamatergic transmission, reducing mEPSCs. Remarkably, LSD1 physiological increase occurs in response to psychosocial stress-induced glutamatergic signaling. Since this mechanism entails neuroLSD1 splicing downregulation, we conclude that LSD1/neuroLSD1 ratio modulation in the hippocampus is instrumental to a negative homeostatic feedback, restraining glutamatergic neuroplasticity in response to glutamate. The active process of forgetting provides memories with salience. With our work, we propose that softening memory traces of adversities could further represent a stress-coping process in which LSD1/neuroLSD1 ratio modulation may help preserving healthy emotional references.

Ultrastructural Evidence for a Role of Astrocytes and Glycogen-Derived Lactate in Learning-Dependent Synaptic Stabilization.

Long-term memory formation (LTM) is a process accompanied by energy-demanding structural changes at synapses and increased spine density. Concomitant increases in both spine volume and postsynaptic density (PSD) surface area have been suggested but never quantified in vivo by clear-cut experimental evidence. Using novel object recognition in mice as a learning task followed by 3D electron microscopy analysis, we demonstrate that LTM induced all aforementioned synaptic changes, together with an increase in the size of astrocytic glycogen granules, which are a source of lactate for neurons. The selective inhibition of glycogen metabolism in astrocytes impaired learning, affecting all the related synaptic changes. Intrahippocampal administration of l-lactate rescued the behavioral phenotype, along with spine density within 24 hours. Spine dynamics in hippocampal organotypic slices undergoing theta burst-induced long-term potentiation was similarly affected by inhibition of glycogen metabolism and rescued by l-lactate. These results suggest that learning primes astrocytic energy stores and signaling to sustain synaptic plasticity via l-lactate.

[On the benefits to keep using the asperger diagnosis]. / De l'intérêt à continuer de poser le diagnostic de Syndrome d'Asperger.

The purpose of this paper is to examine the possible benefits to keep using the diagnosis of Asperger's syndrome. We first describe the evolution of this entity over time and within nomenclature bases such as the ICD- 10, the CFTMEA and the last versions of DSM. Then, we discuss more precisely the impact of the decision made in the DSM-5 to suppress the Asperger syndrome as a differentiated entity within the pervasive developmental disorders (PDD). This disorder chapter by the way also disappears and is replaced by Autism Spectrum Disorder (ASD). We present here three clinical cases encountered in an outpatient general child psychiatry clinic : 1 case was diagnosed as Asperger syndrome, 1 as infantile autism (early infantile autism) and 1 as another pervasive developmental disorder (psychotic disharmony). The objective was to expose the commonali ties and differences between these three entities. We conclude that keeping using the Asperger diagnosis is important for the clinical management of these clinical situations but also for the individual, his or her family and society at large.

L'objet de cet article est de discuter du diagnostic de syndrome d'Asperger. Nous décrivons tout d'abord l'évolution de cette entité au fil du temps et des référentiels que sont l'ICD-10, la CFTMEA et les différents DSM. Nous parlons plus précisément des répercussions des décisions prises dans la nouvelle mouture du DSM-5 pour ce trouble en particulier, à savoir sa disparition en tant qu'entité différenciée au sein des troubles envahissants du développement (TED) dont le terme disparaît également au profit du concept de troubles du spectre autistique (TSA). Nous exposons 3 cas cliniques rencontrés en consultation de pédopsychiatrie générale : 1 cas diagnostiqué comme étant un syndrome d'Asperger, 1 cas d'autisme infantile (autisme infantile précoce) et 1 cas d'un autre trouble envahissant du développement (dysharmonie psychotique) afin d'exposer les points communs et les différences entre ces 3 entités. Nous insistons sur le fait de l'importance de poser ce diagnostic et de le reconnaître en tant qu'entité à part entière au niveau de notre clinique quotidienne non seulement pour la prise en charge de ces situations cliniques mais aussi et surtout pour l'individu, sa famille et la société.

Testing Aß toxicity on primary CNS cultures using drug-screening microfluidic chips.

Open microscale cultures of primary central nervous system (CNS) cells have been implemented in microfluidic chips that can expose the cells to physiological fluidic shear stress conditions. Cells in the chips were exposed to differently aggregated forms of beta-amyloid (Aß), i.e. conditions mimicking an Alzheimer's Disease environment, and treated with CNS drugs in order to assess the contribution of glial cells during pharmacological treatments. FTY720, a drug approved for the treatment of Multiple Sclerosis, was found to play a marked neuroprotective role in neuronal cultures as well as in microglia-enriched neuronal cultures, preventing neurodegeneration after cell exposure to neurotoxic oligomers of Aß.

Autophagy as a new therapeutic target in Duchenne muscular dystrophy.

A resolutive therapy for Duchene muscular dystrophy, a severe degenerative disease of the skeletal muscle, is still lacking. Because autophagy has been shown to be crucial in clearing dysfunctional organelles and in preventing tissue damage, we investigated its pathogenic role and its suitability as a target for new therapeutic interventions in Duchenne muscular dystrophy (DMD). Here we demonstrate that autophagy is severely impaired in muscles from patients affected by DMD and mdx mice, a model of the disease, with accumulation of damaged organelles. The defect in autophagy was accompanied by persistent activation via phosphorylation of Akt, mammalian target of rapamycin (mTOR) and of the autophagy-inhibiting pathways dependent on them, including the translation-initiation factor 4E-binding protein 1 and the ribosomal protein S6, and downregulation of the autophagy-inducing genes LC3, Atg12, Gabarapl1 and Bnip3. The defective autophagy was rescued in mdx mice by long-term exposure to a low-protein diet. The treatment led to normalisation of Akt and mTOR signalling; it also reduced significantly muscle inflammation, fibrosis and myofibre damage, leading to recovery of muscle function. This study highlights novel pathogenic aspects of DMD and suggests autophagy as a new effective therapeutic target. The treatment we propose can be safely applied and immediately tested for efficacy in humans.

When nominal features are marked on verbs: a transcranial magnetic stimulation study.

It has been claimed that verb processing (as opposed to noun processing) is subserved by specific neural circuits in the left prefrontal cortex. In this study, we took advantage of the unusual grammatical characteristics of clitic pronouns in Italian (e.g., lo and la in portalo and portala 'bring it [masculine]/[feminine]', respectively)-the fact that clitics have both nominal and verbal characteristics, to explore the neural correlates of verb and clitic processing. We used repetitive transcranial magnetic stimulation (rTMS) to suppress the excitability of the left prefrontal cortex and to assess its role in producing verb+det+noun and verb+clitic phrases. Results showed an interference effect for both kinds of phrases when stimulation was applied to the left but not to the right prefrontal cortex. However, the interference effect was significantly greater for the verb+clitic than for the verb+det+noun phrases. These findings support the view that clitics increase the morphosyntactic complexity of verbs.

Role of the cerebellum in time perception: a TMS study in normal subjects.

The aim of this study was to investigate the role of the cerebellum in a temporal-discrimination task without movement production in healthy subjects. Ten healthy subjects underwent a time-perception task with somatosensory stimuli. Two pairs of electrical stimuli: the first considered the reference pair (rp) with a standard interval of 400 ms and the second, the test pair (tp), with variable intervals ranging from 300 to 500 ms, were applied by surface electrodes on the right forearm. Subjects were instructed to compare time intervals of rp and tp and to estimate whether the tp interval was shorter than, equal to, or longer than that of rp. The task was performed in baseline and after 1 Hz rTMS over the right and left cerebellar hemisphere. The right cerebellar rTMS worsened temporal discrimination of cutaneous somatosensory electrical stimuli on the ipsilateral hand. rTMS of the left cerebellar hemisphere did not determine significant changes in the subjects' performance with respect to the baseline. These findings suggest that the cerebellum plays a role in merely perceptive aspects of temporal information processing.

Oocyte development and egg envelope formation in Oreochromis niloticus, a mouth-brooding cichlid fish.

The development of the oocyte and of its associated follicle cells in the Nile tilapia, Oreochromis niloticus, has been examined by optical and transmission electron microscopy. During oocyte development the female gamete of Orochromis niloticus increases in size because of the accumulation of yolk in its cytoplasm. As the accumulation of yolk proceeds, the organization of cortex of the oocyte becomes very complex; all of the cytoplasmic organelles and several populations of vesicles can be found. On the other hand follicle cells also undergo a series of modifications: they first become cuboidal then cylindrical and their cytoplasm become densely populated with organelles. The mature egg of Oreochromis niloticus is surrounded by a thin acellular envelope (chorion) assembled during oocyte development. Biochemical analysis of isolated and purified chorions from mature females was also performed. SDS-PAGE under reducing conditions showed a reproducible pattern of three major polypeptides (121, 66 and 50 kD), most of which being glycosylated. The pattern of synthesis and assembly of the egg envelope in Oreochromis niloticus, a mouth-brooding cichlid fish, is also discussed.

Alpha(v)beta3 and alpha(v)beta5 integrin expression in glioma periphery.

OBJECTIVE: This study analyzed the expression of integrins alpha(v)beta3 and alpha(v)beta5 in glioma tissue and focused on the periphery of high-grade gliomas. METHODS: The analysis was performed with Western blot, immunohistochemistry, and immunofluorescence, by use of two monoclonal antibodies able to recognize the functional integrin heterodimer. The expression of integrin-related ligands and growth factors also was studied. Sections from the tumor periphery were classified as either tumor periphery (light tumor infiltrate or scant visible cells) or peritumor (heavy tumor infiltration). RESULTS: Our data on glioma tissues demonstrated that both integrins were expressed in glioma cells and vasculature and their expression correlated with the histological grade. Alpha(v)beta3 expression was prominent in astrocytic tumors. Both integrins were markers of tumor vasculature, particularly of endothelial proliferation. A high-grade glioma periphery demonstrated a prominent expression of integrin alpha(v)beta3. Cells demonstrating alpha(v)beta3 positivity were identified as tumor astrocytes and endothelial cells by double imaging. The same cells were surrounded by some alpha(v)beta3 ligands and co-localized fibroblast growth factor 2. Matrix metalloproteinase 2 also was found to be co-localized with alpha(v)beta3 in the same cells. Alpha(v)beta3 expression was more relevant in tumor astrocytes. Alpha(v)beta3 integrin and vascular endothelial growth factor expression increased from the periphery to the tumor center. CONCLUSION: Our data support the role of integrins alpha(v)beta3 and alpha(v)beta5 in glioma-associated angiogenesis. In addition, they suggest a role for integrin alpha(v)beta3 in neoangiogenesis and cell migration in high-grade glioma periphery.

Effects of purified recombinant neural and muscle agrin on skeletal muscle fibers in vivo.

Aggregation of acetylcholine receptors (AChRs) in muscle fibers by nerve-derived agrin plays a key role in the formation of neuromuscular junctions. So far, the effects of agrin on muscle fibers have been studied in culture systems, transgenic animals, and in animals injected with agrin--cDNA constructs. We have applied purified recombinant chick neural and muscle agrin to rat soleus muscle in vivo and obtained the following results. Both neural and muscle agrin bind uniformly to the surface of innervated and denervated muscle fibers along their entire length. Neural agrin causes a dose-dependent appearance of AChR aggregates, which persist > or = 7 wk after a single application. Muscle agrin does not cluster AChRs and at 10 times the concentration of neural agrin does not reduce binding or AChR-aggregating activity of neural agrin. Electrical muscle activity affects the stability of agrin binding and the number, size, and spatial distribution of the neural agrin--induced AChR aggregates. Injected agrin is recovered from the muscles together with laminin and both proteins coimmunoprecipitate, indicating that agrin binds to laminin in vivo. Thus, the present approach provides a novel, simple, and efficient method for studying the effects of agrin on muscle under controlled conditions in vivo.

Differential activation of NF-kappa B subunits in dendritic cells in response to Gram-negative bacteria and to lipopolysaccharide.

Dendritic cell (DC) maturation is essential for the initiation of T-dependent immune responses. Nuclear factor kappa B/Rel (NF kappa B/Rel) transcription factors are ubiquitously expressed signalling molecules, known to regulate the transcription of a large number of genes involved in immune responses, including cytokines such as IL-1, IL-6, TNF-alpha and cell surface molecules (MHC class I and II, B7.2). In this study, we have compared the activation of five members of the NF-kappa B family, p65, c-Rel, p50, RelB and p52, during DC maturation in response to lipopolysaccharide (LPS) and to Salmonella typhimurium. We have shown that although the translocation of NF-kappa B occurred very early, 30 min after treatment with both S. typhimurium and LPS, bacteria-induced NF-kappa B activation was more pronounced. Four out of five members, i.e. p65, c-Rel, p50 and RelB, were similarly activated upon the two stimuli but with different kinetics. Indeed, we have observed that p65, c-Rel and p50 were translocated early, whereas RelB was translocated later in DC activation. This differential regulation suggests that the various members of NF-kappa B family can mediate distinct functions of DC physiology.

Hexarelin, but not growth hormone, protects heart from damage induced in vitro by calcium deprivation replenishment.

The effects of hexarelin, a growth hormone (GH) secretagogue, and human GH on the mechanical and metabolic changes measured in isolated rat hearts submitted to 5 min of Ca2+ deprivation followed by reperfusion with Ca2+-containing medium, the so-called calcium paradox phenomenon, were studied. Hexarelin (80 microg/kg bid, subcutaneously) administered for 7 d to male rats effectively antagonized the sudden increase in resting tension measured in vitro on Ca2+ repletion. Moreover, during Ca2+ repletion the release of creatine kinase activity (an index of cell damage) in the perfusate of these hearts was reduced up to 40% compared with controls. By contrast, administration of hexarelin for 3 d or GH (400 microg/kg bid, subcutaneously) for 7 d did not affect the mechanical and metabolic alterations induced by the calcium paradox. To assess its direct and acute cardiac effects, hexarelin (8 microg/mL) was perfused in vitro in recirculating conditions for 60 min through the hearts of normal rats. In this case, hexarelin did not stimulate heart contractility and failed to prevent ventricular contracture upon Ca2+ readmission, whereas diltiazem, a Ca2+channel blocker, effectively antagonized the calcium paradox phenomenon. We conclude that short-term in vivo exposure to hexarelin, but not GH, enables cardiac myocyites to prevent cytoplasmatic electrolytic unbalance and to control intracellular Ca2+ gain, two functions largely impaired during the calcium paradox phenomenon. Moreover, because the effect of hexarelin is not acute but dependent on the length of in vivo treatment, we suggest that it requires modifications of myocardiocyte physiology.

Dendritic cells express tight junction proteins and penetrate gut epithelial monolayers to sample bacteria.

Penetration of the gut mucosa by pathogens expressing invasion genes is believed to occur mainly through specialized epithelial cells, called M cells, that are located in Peyer's patches. However, Salmonella typhimurium that are deficient in invasion genes encoded by Salmonella pathogenicity island 1 (SPI1) are still able to reach the spleen after oral administration. This suggests the existence of an alternative route for bacterial invasion, one that is independent of M cells. We report here a new mechanism for bacterial uptake in the mucosa tissues that is mediated by dendritic cells (DCs). DCs open the tight junctions between epithelial cells, send dendrites outside the epithelium and directly sample bacteria. In addition, because DCs express tight-junction proteins such as occludin, claudin 1 and zonula occludens 1, the integrity of the epithelial barrier is preserved.

Ultrastructural features of the gut in the white sturgeon, Acipenser transmontanus.

Electron-microscopic examinations of the sturgeon gut were performed. Oesophageal goblet cells were abundant in the stratified epithelium. The ultrastructural features of the secretory granules of the oesophageal and intestinal goblet cells were quite similar to those of other vertebrates. Lobules of multilocular adipose tissue were observed in the deep tunica propriasubmucosa of the oesophagus, in close association with vasculature and large fibre bundles of myelinated and unmyelinated axons. Similarly composed nerve fibre bundles were observed in the cardiac stomach, too. The presence of myelinated axons is an unusual feature in the vertebrate enteric nervous system. Cardiac and fundic zones of the stomach showed an epithelium with columnar ciliated and non-ciliated cells, the latter equipped with fuzzy microvilli. Cells lining the tubular gastric proper glands were markedly granulated. Intestinal superficial epithelium was columnar and contained ciliated, as well as non-ciliated and goblet cells. In the tunica propria all over the intestine, the presence and ultrastructure of granulated cells was in addition described. Intraepithelial granulated leukocytes were seen throughout the alimentary canal. Various types of endocrine cells were seen both in the stomach and in the intestine, the size of their granules was measured and their ultrastructure described and compared to that of mammalian cell types.

Role of subunit composition in determining acetylcholine receptor degradation rates in rat myotubes.

During neuromuscular junction maturation, the rapidly degrading receptors (Rr; t1/2 approximately equal to 1 day) are replaced by metabolically stable molecules (Rs; t1/2 approximately equal to 10 days). Rr and Rs do not interconvert, are differently regulated after denervation in adult muscle and are endowed of unique responses to stabilizing agents. In cultured rat myotubes all the epsilon subunit-containing acetylcholine receptors (epsilon-AchRs) are of the Rs type. In the present study we show that Rs exist also in absence of epsilon-AChR and that nonepsilon-(presumably gamma-)AChRs can be included in the Rs pool when epsilon-AChR expression is low. The data indicate that Rs metabolic properties are independent of AChR subunit composition and that epsilon subunit is a signal to efficiently sort AChR molecules to the Rs pool.

Metabolism and trafficking of N-type voltage-operated calcium channels in neurosecretory cells.

The N-type voltage-operated calcium channel has been characterized over the years as a high-threshold channel, with variable inactivation kinetics, and a unique ability to bind with high affinity and specificity omega-conotoxin GVIA and related toxins. This channel is particularly expressed in some neurons and endocrine cells, where it participates in several calcium-dependent processes, including secretion. Omega-conotoxin GVIA was instrumental not only for the biophysical and pharmacological characterization of N-type channels but also for the development of in vitro assays for studying N-type VOCC subcellular localization, biosynthesis, turnover, as well as short-and long-term regulation of its expression. We here summarize our studies on N-type VOCC expression in neurosecretory cells, with a major emphasis on recent data demonstrating the presence of N-type channels in intracellular secretory organelles and their recruitment to the cell surface during regulated exocytosis.

The interaction of sperm cells with exogenous DNA: a role of CD4 and major histocompatibility complex class II molecules.

Mouse epidydimal sperm cells have the spontaneous ability to take up exogenous DNA, a part of which is further internalized into nuclei. We report here that sperm cells from MHC class II knockout mice have a reduced ability to bind DNA compared to sperm cells from wild-type animals. Spermatozoa from CD4 knockout mice are instead fully capable of binding exogenous DNA, yet lose the ability to further internalize it. MHC class II expression was not detected on sperm heads using monoclonal antibodies. In contrast, CD4 molecules were found on sperm heads by both immunofluorescence and Western blot analysis. Moreover, we show that nuclear internalization of exogenous DNA was prevented in wild-type sperm cells preincubated with anti-CD4 mAbs. These results support the conclusion that CD4 and MHC class II molecules play distinct roles in the process of sperm/DNA interaction: though not present in mature sperm cells, MHC class II expression appears to be required during spermatogenesis to produce sperm cells capable of taking up foreign DNA, while CD4 molecules present on sperm cells mediate the nuclear internalization of sperm-bound DNA.

The effects of antifungal agents on the morphogenetic transformation by Candida albicans in vitro.

The effects of antifungal agents, with different mechanisms of action, on the morphogenetic transformation by synchronised yeast-phase Candida albicans cells in vitro and their respective anti-Candida activities are described. MIC data demonstrated that the azoles, amphotericin B and echinocandin were the most active agents against four C. albicans strains. Morphogenetic transformation experiments demonstrated that amphotericin B was significantly better at preventing the transformation, under a variety of test conditions, than the azoles and flucytosine: amphotericin B abolished the transformation at low concentrations while the azoles only prevented the morphogenetic transformation at much higher concentrations (> 100 x MIC).

Modulation of microtubule shape in vitro by high molecular weight microtubule associated proteins MAP1A, MAP1B, and MAP2.

The effect of microtubule associated proteins on microtubule shape has been investigated in reconstitution experiments using purified tubulin and purified MAP1A, MAP1B, and MAP2. Microtubules assembled in the presence of these MAPs were fixed with 0.1% glutaraldehyde and, after negative staining, were examined by electron microscopy. The results show that MAP1A microtubules were generally short and "straight' while those assembled with MAP1B were longer and "bendy'. MAP2 microtubules showed both types of morphologies even though straight microtubules were more abundant. These data suggest that MAPs may modulate not only microtubule dynamics but also microtubule shape which may be important in their spatial distribution and/or role in specific neuronal areas.