Short-lived TCR microclusters and a longer-lived protein kinase Ctheta-focusing central supramolecular activation cluster (cSMAC) have been defined in model immunological synapses (IS). In different model systems, CD28-mediated costimulatory interactions have been detected in microclusters, the cSMAC, or segregated from the TCR forming multiple distinct foci. The relationship between TCR and costimulatory molecules in the physiological IS of T cell-dendritic cell (DC) is obscure. To study the dynamic relationship of CD28-CD80 and TCR interactions in the T cell-DC IS during Ag-specific T cell activation, we generated CD80-eCFP mice using bacterial artificial chromosome transgenic technology. In splenic DCs, endogenous CD80 and CD80-eCFP localized to plasma membrane and Golgi apparatus, and CD80-eCFP was functional in vivo. In the OT-II T cell-DC IS, multiple segregated TCR, CD80, and LFA-1 clusters were detected. In the T cell-DC synapse CD80 clusters were colocalized with CD28 and PKCtheta, a
Cholesterol- and glycosphingolipid-enriched membrane lipid microdomains, frequently called lipid rafts, are thought to play an important role in the spatial and temporal organization of immunological synapses. Higher ordering of lipid acyl chains was suggested for these entities and imaging of membrane order in living cells during activation can therefore help to understand the mechanisms responsible for the supramolecular organization of molecules involved in the activation of T cells. Here, we employ the phase-sensitive membrane dye di-4-ANEPPDHQ together with a variety of spectrally-resolved microscopy techniques, including 2-channel ratiometric TIRF microscopy and fluorescence lifetime imaging, to characterize membrane order at the T cell immunological synapse at high spatial and temporal resolution in live cells at physiological temperature. We find that higher membrane order resides at the immunological synapse periphery where proximal signalling through the immunoreceptors and accessory proteins
Although the accumulation of lipid rafts at the immunological synapse is now well accepted, the degree of the accumulation, the localization within the fine structure of the immunological synapse, and the region from which lipid rafts are recruited have not been defined. In this work we show that lipid rafts preferentially accumulate in the central zone of the immunological synapse, the central supramolecular activation complex (C-SMAC). However, quantitative analyses indicate that the level of recruitment of lipid rafts to the C-SMAC is relatively small and suggests that rearrangement of lipid rafts from the peripheral zone of the synapse into the C-SMAC can account for this accumulation. We also assessed the effects of CD28 deficiency on lipid raft recruitment to the immunological synapse. The accumulation of lipid occurred independently of the CD28/B7 system and was not measurably altered by CD28.
Immunological synapses are organized cell-cell junctions between T lymphocytes and APCs composed of an adhesion ring, the peripheral supramolecular activation cluster (pSMAC), and a central T cell receptor cluster, the central supramolecular activation cluster (cSMAC). In CD8+ cytotoxic T lymphocytes, the immunological synapse is thought to facilitate specific killing by confining cytotoxic agents to the synaptic cleft. We have investigated the interaction of human CTLs and helper T cells with supported planar bilayers containing ICAM-1. This artificial substrate provides identical ligands to CD4+ and CD8+ T cells, allowing a quantitative comparison. We found that cytotoxic T lymphocytes form a ring junction similar to a pSMAC in response to high surface densities of ICAM-1 in the planar bilayer. MICA, a ligand for NKG2D, facilitated the ring junction formation at lower surface densities of ICAM-1. ICAM-1 and MICA are upregulated in tissues by inflammation- and stress-associated signaling, ...
Immunological synapses are initiated by signaling in discrete T cell antigen receptor microclusters and are important for the differentiation and effector functions of T cells. Synapse formation involves the orchestrated movement of microclusters toward the center of the contact area with the antigen-presenting cell. Microcluster movement is associated with centripetal actin flow, but the function of motor proteins is unknown. Here we show that myosin IIA was necessary for complete assembly and movement of T cell antigen receptor microclusters. In the absence of myosin IIA or its ATPase activity, T cell signaling was interrupted downstream of the kinase Lck and the synapse was destabilized. Thus, T cell antigen receptor signaling and the subsequent formation of immunological synapses are active processes dependent on myosin IIA.
Our data also support a role for the immunological synapse in the discrimination of potential antigenic ligands by the TCR. The TCR exhibits an exquisite specificity for its antigen as demonstrated by the significantly different biological outcomes induced by ligands that differ by only a single amino acid. Examination of TCR interaction with MHC-peptide by a panel of altered peptide ligands showed a direct relationship between the process of immunological synapse formation and T cell proliferation. Furthermore, all densities of agonist MHC-peptide complexes that induced T cell proliferation also triggered a central cluster with greater than or equal to 60 molecules per square micrometer of accumulated MHC-peptide complexes. As the number of accumulated MHC-peptide complexes was "locked in" in the final stage of immunological synapse formation, the T cell could thus set a specific threshold to determine whether to commit to the program of T cell activation.. Given the discrete temporal stages of ...
Professor Dustin has a B.A. from Boston University (Biology, 1984) and trained with Timothy A. Spring for his Ph.D. (Harvard University, 1990). After post-doctoral training with Stuart Kornfeld at Washington University (1993) he joined the Department of Pathology as an Assistant Professor. There, he led a collaborative team in uncovering the dynamics of the T cell immunological synapse. He moved to the Skirball Institute of Biomolecular Medicine at New York University in 2001, opening studies on the immunological synapse in vivo in settings of vaccination, sterile injury, infection and cancer. He moved to the Kennedy Institute of Rheumatology at the University of Oxford in 2013 with the support of a Wellcome Trust Principal Research Fellowship. The lab is currently focusing on a high throughput approach for immunological synapse characterization and the physiological and clinical relevance of T cell receptor enriched extracellular vesicles generated in the center of the immunological synapse. ...
The specialized junction between a T lymphocyte and an antigen-presenting cell, the immunological synapse, consists of a central cluster of T cell receptors surrounded by a ring of adhesion molecules. Immunological synapse formation is now shown to be an active and dynamic mechanism that allows T cells to distinguish potential antigenic ligands. Initially, T cell receptor ligands were engaged in an outermost ring of the nascent synapse. Transport of these complexes into the central cluster was dependent on T cell receptor-ligand interaction kinetics. Finally, formation of a stable central cluster at the heart of the synapse was a determinative event for T cell proliferation. ...
Immunological synapses arise at contact zones between antigen-presenting cells and T cells. Synapses have distinct organization and are composed of T cell receptors interacting with major histocompatibility complex-peptide entities surrounded by various potentiating coreceptors and signaling molecules that migrate into the synapse through cytoskeleton-dependent processes. Krummel et al. have extended initial studies in lipid bilayers to explore the architecture and the extraordinary dynamics of immunological synapse formation by real-time video microscopy of whole cells containing green fluorescent protein chimeras.. Krummel, M.F., Sjaastad. M.D., Wülfing, C., and David, M.M. (2000) Differential clustering of CD4 and CD3ζ during T cell recognition. Science 289: 1349-1352. [Abstract] [Full Text]. ...
We show here that the CD4 coreceptor is a regulatory molecule that helps to govern both recruitment of the TCR to lipid rafts as well as formation of the immunological synapse. In particular, we show that raft-associated CD4 is critical in regulating TCR/PKCθ clustering at the synapse site. We demonstrate that CD4-deficient Th1 cells are impaired in their ability to cluster the TCR and PKCθ at the site of T/APC contact, indicating that CD4 also plays a role in immunological synapse formation, particularly in response to low-affinity peptide stimulation. Kinetic studies indicate that this inability to cluster the TCR and PKCθ in the absence of CD4 does not simply represent a kinetic delay.. These data enhance our understanding of the ways in which CD4 regulates the formation of the macromolecular TCR complex. We have previously demonstrated a role for CD4 in regulating the association of CD45 and the TCR. The present study implicates CD4 in mediating additional levels of TCR macromolecular ...
Cytolytic cells of the immune system destroy pathogen-infected cells by polarised exocytosis of secretory lysosomes containing the pore-forming protein perforin. Precise delivery of this lethal hit is essential to ensuring that only the target cell is destroyed. In cytotoxic T lymphocytes (CTLs), this is accomplished by an unusual movement of the centrosome to contact the plasma membrane at the centre of the immunological synapse formed between killer and target cells. Secretory lysosomes are directed towards the centrosome along microtubules and delivered precisely to the point of target cell recognition within the immunological synapse, identified by the centrosome. We asked whether this mechanism of directing secretory lysosome release is unique to CTL or whether natural killer (NK) and invariant NKT (iNKT) cytolytic cells of the innate immune system use a similar mechanism to focus perforin-bearing lysosome release. NK cells were conjugated with B-cell targets lacking major histocompatibility
Video articles in JoVE about bilayer include Biomembrane Fabrication by the Solvent-assisted Lipid Bilayer (SALB) Method, Automated Lipid Bilayer Membrane Formation Using a Polydimethylsiloxane Thin Film, Super-resolution Imaging of the Natural Killer Cell Immunological Synapse on a Glass-supported Planar Lipid Bilayer, Lipid Bilayer Vesicle Generation Using Microfluidic Jetting, Gramicidin-based Fluorescence Assay; for Determining Small Molecules Potential for Modifying Lipid Bilayer Properties, Ligand Nano-cluster Arrays in a Supported Lipid Bilayer, Single Molecule Methods for Monitoring Changes in Bilayer Elastic Properties, SNARE-mediated Fusion of Single Proteoliposomes with Tethered Supported Bilayers in a Microfluidic Flow Cell Monitored by Polarized TIRF Microscopy, Reconstitution of a Kv Channel into Lipid Membranes for Structural and Functional Studies, Multifunctional, Micropipette-based Method for Incorporation And Stimulation of Bacterial Mechanosensitive Ion Channels in
T cells are activated when their T-cell antigen receptors (TCRs) are triggered by interactions with antigen peptide-major histocompatibility complex (pMHC) proteins on antigen-presenting cell (APC) surfaces. T cells exhibit exquisite selectivity and sensitivity, and the physical basis for these attributes within the TCR signaling system has attracted much interest. A significant aspect of TCR triggering is the spatial assembly of receptors on multiple length scales. At the scale of the cell itself, TCRs are visibly transported towards the center of the T-cell-APC interface, and the resulting spatial patterns of TCRs and other molecules are called the immunological synapse (Monks et al., 1998; Grakoui et al., 1999). Physically interfering with these transport processes can induce detectable changes in T cell signaling, such as phosphorylation of immunoreceptor tyrosine-based activation motifs on the TCR complex and intracellular calcium flux (Mossman et al., 2005). On shorter time and length ...
Early downstream responses of T lymphocytes following T cell antigen receptor (TCR) activation are mediated by protein complexes that assemble in domains of the plasma membrane. Using stable isotope labeling with amino acids in cell culture and mass spectrometry, we quantitatively related the proteome of αCD3 immunoisolated native TCR signaling plasma membrane domains to that of control plasma membrane fragments not engaged in TCR signaling. Proteins were sorted according to their relative enrichment in isolated TCR signaling plasma membrane domains, identifying a complex protein network that is anchored in the vicinity of activated TCR. These networks harbor widespread mediators of plasma membrane-proximal T cell activities, including propagation, balancing, and attenuation of TCR signaling, immune synapse formation, as well as cytoskeletal arrangements relative to TCR activation clusters. These results highlight the unique potential of systematic characterizations of plasma membrane-proximal T cell
Immunological synapses are organized cell-cell junctions between T lymphocytes and APCs composed of an adhesion ring, the peripheral supramolecular activation cluster (pSMAC), and a central T cell receptor cluster, the ...
TY - JOUR. T1 - The perforin-dependent immunological synapse allows T-cell activation-dependent tumor targeting by MLV vector particles. AU - Kottke, T.. AU - Qiao, J.. AU - Diaz, R. M.. AU - Ahmed, A.. AU - Vroman, B.. AU - Thompson, J.. AU - Sanchez-Perez, L.. AU - Vile, Richard Geoffrey. PY - 2006/8. Y1 - 2006/8. N2 - We have reported that retroviral particles adhered to the surface of antigen-specific T cells can be carried to metastases following adoptive transfer in vivo, a process we have called viral hitch hiking. Following antigen-driven T-cell accumulation at tumors, viral particles productively infect tumor cells via envelope/receptor dependent interactions (hand on of virus from the T cell to the tumor cell). We describe here a second envelope/receptor independent pathway of viral hand on from T cells, dependent on T-cell activation. We show that the endosomolytic property of perforin promotes release of viral particles from endosomes into which they are co-delivered along with ...
Author(s): Krummel, Matthew F.; Cahalan, Michael D. | Abstract: The immunological synapse (IS) as a concept has evolved from a static view of the junction between T cells and their antigen-presenting cell partners. The entire process of IS formation and extinction is now known to entail a dynamic reorganization of membrane domains and proteins within and adjacent to those domains. The entire process is also intricately tied to the motility machinery-both as that machinery directs
The great majority of mammalian genes yield multiple transcripts arising from differential mRNA processing, but in very few instances have alternative forms been assigned distinct functional properties. We have cloned and characterized a new isoform of the accessory molecule CD6 that lacks the CD166 binding domain and is expressed in rat and human primary cells. The novel isoform, CD6Deltad3, results from exon 5 skipping and consequently lacks the third scavenger receptor cysteine-rich (SRCR) domain of CD6. Differential expression of the SRCR domain 3 resulted in a remarkable functional difference: whereas full-length CD6 targeted to the immunological synapse, CD6Deltad3 was unable to localize at the T cell:APC interface during Ag presentation. Analysis of expression of CD6 variants showed that, while being more frequent in coexpression with full-length CD6, the CD6Deltad3 isoform constituted the sole species in a small percentage of T cells. In the rat thymus, CD6Deltad3 is less represented in double
Here, we described the dynamics of distinct assemblies of the functionally connected TCR-proximal signaling components SLP-76, LAT, and ZAP-70 at the IS. The adaptor protein LAT is an integral plasma membrane protein that links antigen recognition by the TCR with several key downstream signaling components (1, 31). High-resolution imaging studies have shown that antigen receptors and LAT are organized in distinct domains at the plasma membrane, which are juxtaposed, but remain separate, upon antigenic stimulation (32, 33). LAT functionality therefore involves the spatiotemporal regulation of protein assemblies. Moreover, a substantial fraction of LAT accumulates within intracellular vesicles (3, 21, 22). We found that the dynamics of LAT in subsynaptic vesicles was strikingly governed by the architecture of the IS. Specifically, the movement of LAT-containing vesicles was confined to tracks between ZAP-70-containing microclusters that represented TCR activation sites. Also, LAT found in ...
The immunological synapse between T cells and the antigen-presenting dendritic cells acts as a locus for T cell activation. Now, Roberto Maldonado, Laurie Glimcher (Harvard School of Public Health, Boston, MA), and colleagues find that this synapse also helps the T cells decide between two different activated, differentiated fates based on the extent of colocalization of receptors at the synapse.. The end products of this decision are the bacteria-fighting Th1 cells and the parasite-fighting Th2 cells. Activation of the interferon-γ receptor (IFNGR) or interleukin 4 receptor (IL-4R) is known to favor Th1 production or Th2 production, respectively.. Now, Glimchers group shows that the IFNGR but not IL-4R colocalizes with the T cell receptor (TCR) at the immunological synapse. The extent of this colocalization is greatest in mice that tend to generate more Th1 cells. IL-4, which favors production of Th2 cells, inhibits the colocalization.. Turning this colocalization correlation into causation ...
Here, we reported the identification of a PIP2-derived signaling amplification loop for the initiation of B cell activation (fig. S7). Specifically, we observed that there is a highly dynamic spatial-temporal change of PIP2 within the immunological synapse during B cell activation: PIP2 is efficiently depleted inside the BCR microclusters but is regenerated outside the BCR microclusters. Both events are important for the sustained initiation of B cell activation. Mechanistically, the hydrolysis of PIP2 inside the BCR microclusters induced a positive feedback mechanism for its synthesis outside the BCR microclusters.. The positive feedback nature of the PIP2-derived amplification loop is achieved by the unique Brownian mobility of PIP2 metabolic products. DAG, the product of PIP2 hydrolysis within the BCR microclusters, exhibits high Brownian mobility, which ensures its efficient interaction with DGKζ outside the BCR microclusters. PA, converted from DAG by DGKζ, drastically facilitates the ...
In this study, we have adopted a proteomic approach to characterize new ligands of the TCR‐signaling ITAM motifs. In this way, we identified the GPCR‐interacting protein β‐Arr1 as a novel direct ligand of the TCR that is recruited to unphosphorylated ITAMs in a manner dependent on TCR triggering. While β‐Arr1 was previously described as a cytoplasmic effector of both GPCRs and RTKs (Hupfeld & Olefsky, 2007), this is the first demonstration of β‐Arr1 binding to the TCR, a multi‐subunit receptor without intrinsic enzymatic activity. We found that β‐Arr1 recruitment to the TCR was induced by TCR triggering. However, in contrast to the recruitment of other TCR signaling effectors, such as the tyrosine kinase ZAP‐70 or the adaptor protein Nck (Gil et al, 2002), that is mediated by changes in the TCR itself, recruitment of β‐Arr1 to the TCR was mediated by modifying β‐Arr1. The mechanism by which TCR triggering induces the recruitment of β‐Arr1 to non‐triggered TCRs ...
The authors regret that in the original version of their paper, the middle blot for Rac2 in Fig. 2 C was incorrect. The legend remains correct, and the corrected figure appears below. The online HTML and PDF versions of this article have been corrected. The error only remains in the print version.. ...
Lymphocyte dynamics in lymph nodes are governed by environmental GO signals and STOP signals that interact with intrinsic polarity networks in the cell. Chemoki...
The general goal of this laboratory is to elucidate the mechanism of activation, differentiation and function, and its regulation of T cells which play central roles in the regulation of immune responses and homeostasis. These include the regulation of immunological synapse, T cell receptor-induced signal transduction, co-stimulation regulation and signaling in T cell development. Particularly, the mechanism to induce autoimmune diseases and inflammations caused by aberrant regulation of T cell function and activation will be analyzed to aim to overcome the diseases.. ...
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PubMed Central Canada (PMC Canada) provides free access to a stable and permanent online digital archive of full-text, peer-reviewed health and life sciences research publications. It builds on PubMed Central (PMC), the U.S. National Institutes of Health (NIH) free digital archive of biomedical and life sciences journal literature and is a member of the broader PMC International (PMCI) network of e-repositories.
A unique feature of Hodgkin lymphoma (HL) is the presence of CD4+ T cells that surround, protect and promote survival of tumor cells. The adhesion molecules involved in this so-called T cell rosetting are important components of the immunological synapse (IS). However, it is unknown whether this synapse is fully assembled and leads to T cell activation by enabling interaction between the T cell receptor (TCR) and human leukocyte antigen class II (HLA-II). We established a novel rosetting model by co-culturing HLA-II matched PBMCs with HL cell lines and show IS formation with activation of rosetting T cells. HLA-II downregulation by CIITA-knockout did not affect the extent of rosetting, but almost completely abrogated T cell activation. Intriguingly, the level of CD58 expression correlated with the extent of rosette formation and CD58-knockout or CD2 blockade reduced both rosette formation and T cell activation. Extension of our findings to primary HL tissue by immunohistochemistry and proximity ...
Antigen recognition by CD4+ T cells leads to large-scale spatial and temporal molecular redistributions, forming the immunological synapse. We have previously shown that upon dissociation, T cells capture large membrane fragments from antigen-presenting cells directly from the immunological synapse. The mechanism and biological significance of this process, termed trogocytosis, is still unclear. In this thesis I examined the impact that trogocytosis has on the individual T cell after capturing molecules from the antigen presenting cell. I employed murine fibroblast cell lines expressing an I-Ek molecule loaded with a covalently attached antigenic peptide (moth cytochrome C 88-103) and with or without a GFP-tagged cytoplasmic tail as antigen presenting cells for T cells from a peptide-specific TCR transgenic mouse. Using a combination of high-resolution microscopy and flow cytometry, In this thesis I showed that the trogocytosed material is retained on the surface of the T cell and is associated with
Raindrop; Synapse; Developers. synapse x cracked, synapse x serial key, synapse x cracked 2019, synapse x cracked 2020, synapse x serial key generator, synapse x password, synapse x download latest version, synapse x trolling, synapse x register serial key, synapse x discord, synapse x андроид, synapse x авакин лайф, synapse x lumber tycoon, synapse x. Hello all, Ive written a decompiler for Lua 5. This tool does include a great anti detect and anti ban system with built in Proxy and VPN support. Pre-compiled Lua libraries and executables are available at LuaBinaries. SYNAPSE X LUAU DECOMPILER IS HERE! [HOW TO USE] - Duration: 4:09. ProtoSmasher has always been developed with security in mind, making sure youre always undetected and banproof. Synapse Design is the leading SOC design services company, offering design consulting and services in digital, analog/mixed-signal design and software development. Have Protosmasher automatically add a shortcut in windows start menu. ...
Cancer immune evasion is achieved through multiple layers of immune tolerance mechanisms including immune editing, recruitment of tolerogenic immune cells, and secretion of immunosuppressive cytokines. Recent success with immune checkpoint inhibitors in cancer immunotherapy suggests a dysfunctional immune synapse as a pivotal tolerogenic mechanism. Tumor cells express immune synapse proteins to suppress the immune system, which is often modulated by epigenetic mechanisms. When the methylation status of key immune synapse genes was interrogated, we observed disproportionately hypermethylated costimulatory genes and hypomethylation of immune checkpoint genes, which were negatively associated with functional T cell recruitment to the tumor microenvironment. Therefore, the methylation status of immune synapse genes reflects tumor immunogenicity and correlates with survival.. ...
In this review, the putative targets of anti-inflammatory n-3 PUFA, activated during early and late events of T-cell activation will be discussed. Studies have demonstrated that these fatty acids alter plasma membrane micro-organization (lipid rafts) at the immunological synapse, the site where T-cells and antigen-presenting cells (APC) form a physical contact for antigen initiated T-cell signaling. In addition, the production of diacylglycerol and the activation of different isoforms of protein kinase C (PKC), mitogen-activated protein kinase (MAPK), calcium signaling, and nuclear translocation/activation of transcriptional factors, can be modulated by n-3 PUFA ...
Probably involved in the organization of the actin cytoskeleton by acting downstream of CDC42, inducing actin filament assembly. Alters CDC42-induced cell shape changes. In activated T-cells, may play a role in CDC42-mediated F-actin accumulation at the immunological synapse. May play a role in early contractile events in phagocytosis in macrophages.
R. A. Maldonado, Soriano, M. A., L. Perdomo, C., Sigrist, K., Irvine, D. J., Decker, T., and Glimcher, L. H., "Control of T helper cell differentiation through cytokine receptor inclusion in the immunological synapse", Journal of Experimental Medicine, vol. 206, no. 4, pp. 877 - 892, 2009. ...
Part of the first shipment I ever received from Tom Bihn was the Synapse (now called the Synapse 19). I bought it alongside the Aeronaut (now called the Aeronaut 45) and the Co-Pilot. The Synapse has been over the last four years my go-to bag. While I take my Smart Alec to work every day because I need extra space in my bag, the Synapse is the bag I grab if Im going out for the day whether it be in the city or out in the country. The reason I always reach for the Synapse is the same reason people always tell me they wouldnt want a Synapse (19) - because its small. Ive bought a lot of big backpacks over the years, and I never want to carry them because theyre too big. Its the size of the Synapse that makes it so useful. Its never too big, and I always manage to fit more into it than I would imagine that I could.. The Synapse has a pretty spacious main compartment with a stretchy inside pocket thats good for keeping things from mushing up against your back. There are two side pockets. I ...
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Our friends at Razer have launched their Synapse 2.0 beta--and you can have a chance to join by signing up at www.razerzone.com/synapse2. Synapse 2.
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Synapses conduct impulses in our brains. The Cannondale Synapse has fast, electric performance. With a shocking price tag to match, the Synapse is a dream!
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STAMFORD, Conn., March 30, 2011 /PRNewswire/ -- Fujifilm Highlights Synapse® 3D at ACC. Advanced diagnostic applications provide clinical, workflow and...
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行動神経科学、行動神経内分泌学、分子生物学、その他SYNAPSE Lab. としての活動など. 2015.03.11 HPをリニューアルしました。. 研究室 ...
I realize that there are mixed views on this, so I hesitate to ask...but has anyone removed the TB label from a Synapse 19? Im not a huge fan of it a
Every time i try to launch i get a black screen and then this error. Tried setting compatibillty settings and run as admin - no difference. Anyone running the latest ATI Catalyst drivers having the same problem ...
行動神経科学、行動神経内分泌学、分子生物学、その他SYNAPSE Lab. としての活動など. 2015.03.11 HPをリニューアルしました。. 研究室 ...
Synapses are the basis of neural communication. Learn about the types of synapses, what they do, parts of a synapse and how it affects...