Neuroscience. Publications List Edition

Transcription

1 Neuroscience Publications List 3 Edition

2 What is High Content Screening (HCS)? High content screening (HCS), also known as high content analysis, image cytometry, quantitative cell analysis or automated cell analysis, is an automated method that is used to identify substances that alter the phenotype of a cell in a desired manner. This technology is primarily used in biological research and drug discovery and combines fluorescent microscopy, automated cell calculations, and phenotyping using image processing algorithms and informatics tools for the user to make decisions about a treatment. Thermo Scientific High Content Products The portfolio of High Content Analysis products includes assay development and screening tools like the Thermo Scientific ArrayScan XTI High Content Analysis (HCA) Reader and the Thermo Scientific CellInsight NXT High Content Screening (HCS) Platform. Multiple tools are available for assay development on the ArrayScan XTI HCA Reader like the Thermo Scientific X large field-of-view, high resolution camera; Live Cell Chamber; and the new Confocal Module, while our software products like Thermo Scientific HCS Studio Software enable users to develop and make decisions about our assays. With the Thermo Scientific HCS Class and the diverse portfolio of reagents and consumables, we enable scientists to increase their efficiency with their platform while generating more knowledge about the cell. More knowledge evolved into products More out-of-the-box reagents validated for high content More flexibility in instrumentation to address new assay needs Validated SOlutions Information more Knowledge More knowledge about cellular information More measurements and data about cells and their response More information than other cell-based assays Contextual Relevance More knowledge about cells in their context More information in the context of a living cell More tools to characterize complex biologies More knowledge in how to execute HCA More technical resources focused on high content More experience in using, developing, and executing on high content assays Experienced Execution Scientific Validity Scientific validity through literature More peer-reviewed publications in the most relevant and respected journals Automated solutions to increase throughput For more information on Thermo Scientific High Content Products, and a full list of applications by application area, go to thermoscientific.com/highcontent

3 Monitoring Morphology and Synapse Formation in Primary Neurons for Neurotoxicity Assessments and Drug Screening Suk J. Hong and Richik N. Ghosh Thermo Fisher Scientific Pittsburgh, PA USA Application Notes C-AN_NT tamate [µm nate [µm ynaptic intensity synaptic intensity Abstract Synapse formation during nervous system development and degeneration in the pathogenesis of human neurological diseases are highly regulated processes. Subtle changes in the environment of the complex neuronal network may cause either breakdown or creation of synaptic connections. Drug discovery screening for neurological diseases and compound neurotoxicity evaluation would benefit from robust, automated, quantitative in vitro assays that monitor neuronal function. We hypothesized that () toxic insults to the nervous system will cause neuronal synapses to deteriorate in the early phase of neurotoxicity, eventually leading to neurite degeneration and neuronal cell death if the damage is severe; and () an in vitro functional assay for synapse formation and neuronal morphology could be used to monitor and identify such neurotoxic events. We thus developed an automated, functional, high-content screening imaging assay to track and quantify the dynamic changes in neurite morphology and synapses. This assay identifies primary neuronal cells by a neuron-specific and detects synapses on the spines of neurites with pre- and postsynaptic s. The multiplexed targets, including a nuclear, are simultaneously detected with four fluorescent colors, and the fluorescent images of the labeled neurons and synapses are acquired by an automated imaging instrument. The phenotypic features of neuronal morphology and the synapse are automatically identified and quantified on-the-fly by specialized image-analysis software. Such features are potential indicators for neuronal development, differentiation and neurotoxicity, and we could quantify changes in these features under different conditions and for different drug treatments. By monitoring changes in these features, we could also quantitatively evaluate compounds involved in developmental neurotoxicity. In summary, this assay facilitates automation and streamlining of a laborious process in drug discovery screening and compound neurotoxicity assessments; it enables quantitative comparisons between compounds Glutamate [µm in neuronal morphology and function, particularly in neurite and synapse associated events. Kainate [µm O [µm H O [µm ynaptic intensity synaptic intensity ncl [µm ynaptic intensity synaptic intensity vesicl spots vesicl spots ZnCl [µm U6 [µm 6 [µm vesicl spots Introduction Neurons in central and peripheral nervous systems function to transmit electric signals from one location to the other to keep the brain and the body functioning properly. One of the critical structures in the neuron to maintain their proper functional network is synapse, which is the junction between a nerve cell and the cell that receives an impulse from the neuron. The molecular network between these synapses controls not just synaptic signal transmission and synaptic plasticity but also regulates neuronal growth, differentiation and death. The microstructure of synaptic junctions has been extensively studied to understand the relationship between synaptic activity and neuropathophysiology, as well as the molecular mechanism involved in synaptogenesis and the regulation of synapse. Once synaptic function is disrupted by natural or man-made neurotoxic substances, it could lead to long-lasting and often irreversible neuronal damage. Synaptic damage has often been recognized as the first sign of neurodegeneration in many different pathological conditions, including traumatic nerve injury, ischemic stroke, and many neurodegenerative disorders such as Motor Neuron Disease, Alzheimer s, Parkinson s and Huntington s diseases. Many synaptic proteins play an important role in the progression of neurodegenerative diseases. For example, Amyloid beta precursor protein and Presenilin, alpha-synuclein, Huntingtin, Ataxin-, Frataxin and Prion protein are all involved in presynaptic or post-synaptic structure of the neuron and play a role in synaptic damage and neurodegeneration. To measure the synaptic changes that occur in synaptogenesis or synaptic damage, we needed to develop a reliable, accurate, and efficient method to measure accurate synaptic loss, neurite changes and neuronal death. Here we introduce a new way of measuring synaptic function utilizing the power of automated, quantitative, high-content cell-based imaging and analysis. The Thermo Scientific Synaptogenesis HCS Reader HCS Assay Reagents combined with the Thermo Scientific ArrayScan High Content Screening (HCS) Reader and Neuronal Profiling BioApplication enables the quantitation of neuronal morphology and synapses in vitro. On-the-fly automated image analysis and quantitation accompanying the automated image acquisition is done by the Neuronal Profiling BioApplication, which is an automated image analysis software module on the ArrayScan VTI HCS Reader. Using this technology and assay method, we could identify synaptic changes over time and measure synaptic and neurite parameters in an automated manner. Synaptogenesis Assay Target Candidates : EntityTargeted: Candidates for Target: (best assay target screened in bold) Dye & Color: Nucleus Cell Body / Mask MAP-, DyL detected in four different colors. 3 Spot Number per Length 3 3 PSD9, drebrin, spinophilin/neur DyL9 synapsin, syntaxin, DyL69 Table : Potential synaptogenesis HCS assay targets can be DIV DIV 7 6

4 Automated Image Acquisition Automated Plate Delivery ArrayScan VTI HCS Reader Automated Image Acquisition ortical DIV s Whole n (red) physin Real Time Quantitative Image Analysis (BioApplications) Real Time Quantitative Image Analysis (BioApplications) on Raw Image an HCS use cortical rons DIV omics Whole Stain (red) Automated Data Decisions Management aptophysin Analysis & Visualization en) ged on 3 : ayscan HCS Pre-Synaptic Marker, Synaptophysin, Whole campal der DIV Cell Stain andnucleus MAP- Cell Body / en) Raw Image Marker Syna Neuronal Marker MAP- Real Time Quantitative Image Analysis (BioApplications) ArrayScan VTI HCS Reader Neuronal Marker, MAP- (white) Trace from Cell Body (Blue) Automated Data WCS Management PSD9, drebrin, synapsin, Candidates for Target: & Visualization (bestanalysis assay target screened in bold) Discovery ToolBox vhcs Figure : Mouse cortical neurons ( DIV) were stained for synaptophysin (red) and MAP- (green) (left panel), and analyzed (right panel) with the ArrayScan VTI HCS Reader and the Neuronal Profiling v3. BioApplication. Automated Image Acquisition Automated Plate Delivery Marker, PSD-9 (magenta) Co-localization (green) of PSD-9 and Synapt MAP-, (Thermo Scientific Store) Mouse cortical spinophilin/neur syntaxin, neurons DIV Automated, Simultaneous Measurement of Automated Vesicles, Structure Automated Image Acquisition Plate Delivery Cellomics Whole ArrayScan VTI HCS Reader and s Cell Stain (red) Real Time Quantitative Image Analysis (BioApplications) Aβ- Toxicity on Primary Hippocampal Neuron (DIV) DyL9WCS DyL69 DyL Automated Data Real Time Quantitative Image Synaptophysin Decisions Automated Image Acquisition Management (BioApplications) Thermo 3Analysis (green) Scientific HCSExplorer Mouse cortical vesicles intensity (Thermo Scientific Store) spots Imaged on intensity Width neurons DIV Analysis & Visualization DIV 3 ArrayScan HCS DIV vhcs Discovery ToolBox Cellomics Whole 3 Marker Reader Cell Body / Cell Stain (red) Mask Synaptophysin 6 3 Aβ - µm Aβ - Marker, PSD-9 (magenta) of PSD-9 and Synapt µm Co-localization (green) µm Aβ Neuronal Marker, MAP- (white) Trace Aβ from Cell µmbody (Blue) Maximum 9 % 7 (green) Imaged on ArrayScan HCS synapsin, Reader 7 : Markerdrebrin, PSD9, 6 Spot Number per Length MAP-, Dye & Color: A. ucleus A. A. Analyzed Image Branch point (white) Localized Synaptophysin (purple) Neuronal trace (blue) nnel Raw Image vhcs Discovery ToolBox Mask Branch point (white) Localized Synaptophysin (purple) Neuronal trace (blue) Marker Synaptophysin (red) Neuronal Marker MAP- (green) (Thermo Scientific Store) EntityTargeted: Analyzed Image Marker Synaptophysin (red) Neuronal Marker MAP- (green) Aβ - µm ysin (red) ArrayScan er Thermo Scientific HCSExplorer Real Time Quantitative Image Analysis (BioApplications) Automated Image Acquisition Automated Plate Delivery ArrayScan VTI HCS Reader Appl i cati o n No tes C- AN_ NT Automated Measurement of Vesicles and s Using The Thermo Scientific Neuronal Profiling V3. The Thermo Scientific HCS Platform Seamless Integration of all the Steps in Analysis Automated Data 3 6 DyL69 7 DIV 6 DIV Aβ - µm DyL69 synapsin, syntaxin, DyL9 Marker, PSD-9 (magenta) Co-localization (green) of PSD-9 and Synapt Aβ - µm Figure : Mouse cortical neurons were cultured for DIV or DIV and stained for PSD- 9 and MAP-, imaged and analyzed. Only postsynaptic spots stained with numberincreases in Intensity compared tobranch Point PSD-9Neuron antibody DIV neurons DIV Count Total Length spots show neurons (Student s t-test, p<). no Width significant change. er Length 3 Aβ- Toxicity on Primary Hippocampal Neuron Figure : A. Synaptophysin is a good presynaptic. and Whole Cell Stain is used to detect the nuclei and the fine structure of neurites, respectively (Mouse cortical neuron, DIV). DIV DyL DyL69 DIV B. Synaptophysin and MAP- staining for presynaptic vesicle and 3 DIV neuron, DIV). neurite detection (Rat hippocampal DIV 3 9 syntaxin, intens inten Synaptophysin (red) Imaged on the ArrayScan VTI HCS Reader 3 Dye & Color: MAP-, PSD9, drebrin, synapsin, spinophilin/neur syntaxin, Rat Hippocampal Neurons DIV Map- (green) DyL DIV Neuronal Marker, MAP- (white) Trace from Cell Body (Blue) DyL 6 DIV 3 Spot Number per Spot Length Number per Length Cell Body / Mask 3 MAP- 7 7 Spot Number per Length spots (green spot). Co-localized spots (green)represent the Rat Hippocampal Target: spinophilin/neur synapses. neurons DIV location of potential (best assay target Cell Body / (Thermo Scientific Store) Nucleus screened in bold) Map (green) Mask EntityTargeted: Synaptophysin (red) DyL DyL9 DyL69 Imaged on ArrayScan Aβ - µm Aβ µm µm Aβ -DyL9 HCS Reader Candidates for - DyL PunctatedDye PSD-9 Stain Increases by MAP-, PSD9, drebrin, & Color: Target: spinophilin/neur Maturation of Neurons (best assay target screened in bold) Automated Data Management Dye & Color: MAP- Analysis & Visualization vhcs Discovery ToolBox ucleus annel Spot Number per Length Decisions spinophilin/neur syntaxin, Management Hippocampal Thermo Scientific HCSExplorer (Thermo Scientific Store) Analysis & Visualization ons DIV vhcs Discovery ToolBox Cell Body / Nucleus (green) Mouse corticalentitytargeted: Raw Image Analyzed Image Mask neurons DIV Marker Synaptophysin (red) Branch point (white) B. Neuronal Marker, MAP- (w Cell Stain ptophysin (red) Thermo Scientific Whole (red)marker Neuronal Marker MAP- (green) Localized Synaptophysin (purple) Trace Aβ Toxicity on Primary Hippocampal Neuron Automated Data trace (blue) 3 from Cell Bod Decisions -Neuronal DyL DyL9 DyL69 Management : Synaptophysin (green) ed on ArrayScan Thermo Scientific HCSExplorer (Thermo Scientific Store) Rat Hippocampal & Visualization Figure 3: Rat hippocampal neurons ( DIV) were Analysis stained for VTI Reader Imaged on the ArrayScan Candidates forhcs Reader neurons DIV vhcs Discovery ToolBox Body / MAP-, PSD9, drebrin, synapsin, PSD-9 and MAP-, imaged andcell analyzed. Nucleus Target: Map (green) intens EntityTargeted: Mask spinophilin/neur syntaxin, Left panel: detection with MAP- staining. B. Synaptophysin (red) Count inten Total Length Width (best assay target B. DIV Marker Right panel: spot detection with PSD-9 3 Imaged on ArrayScan 3 screened indivbold) spots)for and co-localization with synaptophysin : HCS Reader staining (magenta Candidates 3 MAP-, PSD9, drebrin, synapsin, 9 3

5 resynaptic Marker Rat Hippocampal neurons DIV Cell Body / Map (green) Nucleus and Synapse EntityTargeted: Changes Mask Neurotoxicity Synaptophysin (red) Response Imaged ArrayScan Against Drug Treatments A Glutamate B Kainate C H O D Zinc E U6 HCS Reader Glutamate [µm Width intensity intensity vesicl spots [µm Glutamate [µm Glutamate [µm Glutamate Glutamate [µm Width intensity intensity vesicl spots Kainate [µm Kainate [µm Kainate [µm Kainate [µm Kainate [µm : Candidates for Target: (best assay target screened in bold) Dye & Color: Width Width Width MAP-, DyL 3 PSD9, drebrin, spinophilin/neur DyL9 intensity intensity intensity intensity intensity intensity synapsin, syntaxin, DyL69 vesicl spots H O [µm H O [µm H O [µm H O [µm H O [µm vesicl spots ZnCl [µm ZnCl [µm ZnCl [µm ZnCl [µm ZnCl [µm vesicl spots U6 [µm U6 [µm U6 [µm U6 [µm U6 [µm Spot Number per Length 3 3 Figure : Mouse, rat cortical or hippocampal primary neurons were cultured for DIV, and the dose dependent responses of drugs towards various properties of these neurons were investigated. (A) Glutamate with mm glycine in HBSS was treated for 3 min, washed and replaced with culture media. After hr incubation, neurons were fixed, stained and analyzed. (B) Kainate, (C) H O, (D) Zinc, (E) U6 were treated for hrs in culture media. (Student s t-test, p<, p<, p<). DIV DIV and Synapse Changes as Neurotoxicity Neuronal Marker, MAP- (white) Marker, PSD-9 (magenta) Trace from Cell Body (Blue) Co-localization (green) of PSD-9 and Synapt Response Against Aβ - Aggregates 6 Aβ - µm 6 3 Aβ - Toxicity on Primary Hippocampal Neuron (DIV) Width 6 Aβ - µm Aβ - µm 6 3 intensity intensity Aβ - µm Aβ - 6 HCS Reader vesicles spots Figure 6: Rat hippocampal primary neurons were cultured for DIV. Dose dependent responses of Aβ - aggregates were investigated. mm Aβ - was incubated at 37 C in media for 3 days to induce oligomerization. Neurons were incubated with the Aβ - oligomers for hrs, and then fixed, stained, and analyzed. Aβ - toxicity leads to synapse loss. (Student s t-test, p<, p<, p<). Summary Multiparameter Synaptogenesis Assay simultaneously identifies and quantifies neurites, pre- and post-synaptic structures and synapse in an automated manner. Neurotoxicity from neurotoxic substances is accurately detected. Substances only affecting synapse can be detected. Assay works for acute or chronic neurodegenerative disease cell models. µm 6 3 Spot Number per Length 3 3 DIV DIV Application Notes C-AN_NT thermoscientific.com/highcontent 3 Thermo Fisher Scientific Inc. All rights reserved. Alexa Fluor is a trademark of Molecular Probes, Inc. Packard is a trademark of Perkin Elmer Inc. X-Light is a trademark of Crisel Electrooptical Systems & Technology s.r.l. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details. USA info.cellomics@thermofisher.com Asia info.cellomics.asia@thermofisher.com Europe +3 ()3 7 info.cellomics.eu@thermofisher.com C-AN_NT

6 Neuroscience Publications List Bajenaru, M.L. et al. β Integrin-Focal Adhesion Kinase (Fak) Signaling Modulates Retinal Ganglion Cells Survival. Invest. Ophthalmol. Vis. Sci. 3, 3- (). Bao, R. et al. Targeting Heat Shock Protein 9 with Cudc-3 Overcomes Erlotinib Resistance in Non-Small Cell Lung Cancer. Mol. Cancer Ther., (9). Bauer, P.O. et al. Inhibition of Rho Kinases Enhances the Degradation of Mutant Huntingtin. J. Biol. Chem., (9). Blackmore, M.G. et al. High Content Screening of Cortical Neurons Identifies Novel Regulators of Axon Growth. Mol. Cell. Neurosci., 3- (). Blackmore, M.G. et al. Krüppel-Like Factor 7 Engineered for Transcriptional Activation Promotes Axon Regeneration in the Adult Corticospinal Tract. Proc. Natl. Acad. Sci. U. S. A. 9, 77-7 (). Borikova, A.L. et al. Rho Kinase Inhibition Rescues the Endothelial Cell Cerebral Cavernous Malformation Phenotype. J. Biol. Chem., 7-76 (). Breier, J.M., Radio, N.M., Mundy, W.R. & Shafer, T.J. Development of a High-Throughput Screening Assay for Chemical Effects on Proliferation and Viability of Immortalized Human Neural Progenitor Cells. Toxicol. Sci., 9-33 (). Brickelmaier, M. et al. Identification and Characterization of Mefloquine Efficacy against Jc Virus in Vitro. Antimicrob. Agents Chemother. 3, -9 (9). Buchser, W.J., Pardinas, J.R., Shi, Y., Bixby, J.L. & Lemmon, V.P. 96-Well Electroporation Method for Transfection of Mammalian Central Neurons. BioTechniques, 69-6 (6). Buchser, W.J., Slepak, T.I., Gutierrez-Arenas, O., Bixby, J.L. & Lemmon, V.P. Kinase/Phosphatase Overexpression Reveals Pathways Regulating Hippocampal Neuron Morphology. Mol. Syst. Biol. 6, 39 (). Chantong, B., Kratschmar, D.V., Nashev, L.G., Balazs, Z. & Odermatt, A. Mineralocorticoid and Glucocorticoid Receptors Differentially Regulate NF-κB Activity and Pro-Inflammatory Cytokine Production in Murine BV- Microglial Cells. J. Neuroinflammation 9, (). Clement, C.M., Dandepally, S.R., Williams, A.L. & Ibeanu, G.C. A Synthetic Analog of Verbenachalcone Potentiates Ngf-Induced Outgrowth and Enhances Cell Survival in Neuronal Cell Models. Neurosci. Lett. 9, 7-6 (9). Culbert, A.A. et al. Mapk-Activated Protein Kinase Deficiency in Microglia Inhibits Pro-Inflammatory Mediator Release and Resultant Neurotoxicity: Relevance to Neuroinflammation in a Transgenic Mouse Model of Alzheimer Disease. J. Biol. Chem., (6). Dahl, J.P. et al. Characterization of the Wave Knock-out Mouse: Implications for Cns Development. J. Neurosci. 3, (3). del Valle, K., Theus, M.H., Bethea, J.R., Liebl, D.J. & Ricard, J. Neural Progenitors Proliferation Is Inhibited by Ephb3 in the Developing Subventricular Zone. Int. J. Dev. Neurosci. 9, 9- (). Denis, J.A. et al. Mtor-Dependent Proliferation Defect in Human ES-Derived Neural Stem Cells Affected by Myotonic Dystrophy Type. J. Cell Sci. 6, (3). Doi, H. et al. Rna-Binding Protein Tls Is a Major Nuclear Aggregate-Interacting Protein in Huntingtin Exon with Expanded Polyglutamine-Expressing Cells. J. Biol. Chem. 3, 69-6 ().

7 Doumanis, J., Wada, K., Kino, Y., Moore, A.W. & Nukina, N. Rnai Screening in Drosophila Cells Identifies New Modifiers of Mutant Huntingtin Aggregation. PLoS One (9). Draghetti, C. et al. Functional Whole-Genome Analysis Identifies Polo-Like Kinase and Poliovirus Receptor as Essential for Neuronal Differentiation Upstream of the Negative Regulator αb-crystallin. J. Biol. Chem., (9). Drew, K.L., McGee, R.C., Wells, M.S. & Kelleher-Andersson, J.A. Growth and Differentiation of Adult Hippocampal Arctic Ground Squirrel Neural Stem Cells. J. Vis. Exp. (). Du, W., Hozumi, N., Sakamoto, M., Hata, J. & Yamada, T. Reconstitution of Schwannian Stroma in Neuroblastomas Using Human Bone Marrow Stromal Cells. Am. J. Pathol. 73, 3-6 (). Ehrnhoefer, D.E. et al. A Quantitative Method for the Specific Assessment of Caspase-6 Activity in Cell Culture. PLoS One 6 (). Emery, A.C. & Eiden, L.E. Signaling through the Neuropeptide Gpcr Pac Induces Neuritogenesis Via a Single Linear Camp- and Erk-Dependent Pathway Using a Novel Camp Sensor. FASEB J. 6, (). Emery, A.C., Eiden, M.V. & Eiden, L.E. A New Site and Mechanism of Action for the Widely Used Adenylate Cyclase Inhibitor SQ,36. Mol. Pharmacol. 3, 9- (3). Eva, R., Bouyoucef-Cherchalli, D., Patel, K., Cullen, P.J. & Banting, G. Ip3 3-Kinase Opposes Ngf Driven Outgrowth. PLoS One 7 (). Falsig, J., Porzgen, P., Lotharius, J. & Leist, M. Specific Modulation of Astrocyte Inflammation by Inhibition of Mixed Lineage Kinases with Cep-37. J. Immunol. 73, (). Fennell, M., Chan, H. & Wood, A. Multiparameter Measurement of Caspase 3 Activation and Apoptotic Cell Death in Nt Neuronal Precursor Cells Using High-Content Analysis. J. Biomol. Screen., 96-3 (6). Foley, N.H. et al. Micrornas a and b Are Potent Inducers of Neuroblastoma Cell Differentiation through Targeting of Nuclear Receptor Corepressor. Cell Death Differ., 9-9 (). Fowler, A. et al. A Nonradioactive High-Throughput/High-Content Assay for Measurement of the Human Serotonin Reuptake Transporter Function in Vitro. J. Biomol. Screen., 7-3 (6). Furukawa, Y., Kaneko, K., Yamanaka, K., O Halloran, T.V. & Nukina, N. Complete Loss of Post-Translational Modifications Triggers Fibrillar Aggregation of Sod in the Familial Form of Amyotrophic Lateral Sclerosis. J. Biol. Chem. 3, (). Gao, Y. et al. Inhibition of Y-Box Binding Protein- Slows the Growth of Glioblastoma Multiforme and Sensitizes to Temozolomide Independent O6-Methylguanine- Methyltransferase. Mol. Cancer Ther., (9). Gaublomme, D., Buyens, T. & Moons, L. Automated Analysis of Outgrowth in Mouse Retinal Explants. J. Biomol. Screen., (). Gaughwin, P., Ciesla, M., Yang, H., Lim, B. & Brundin, P. Stage-Specific Modulation of Cortical Neuronal Development by Mmu-Mir-3. Cereb. Cortex, 7-69 (). ge, Y. et al. Promoting Retinal Ganglion Cell Axonal Regeneration by Inhibition of Transforming Growth Factor Beta (Tgf{Beta}) Signaling. Invest. Ophthalmol. Vis. Sci., 67- (). Gies, E. et al. Niclosamide Prevents the Formation of Large Ubiquitin-Containing Aggregates Caused by Proteasome Inhibition. PLoS One ().

8 Guan, J.S. et al. Hdac Negatively Regulates Memory Formation and Synaptic Plasticity. Nature 9, - (9). Hahn, A.T., Jones, J.T. & Meyer, T. Quantitative Analysis of Cell Cycle Phase Durations and PC Differentiation Using Fluorescent Biosensors. Cell Cycle, - (9). Hansen, C. et al. A-Synuclein Propagates from Mouse Brain to Grafted Dopaminergic Neurons and Seeds Aggregation in Cultured Human Cells. J. Clin. Invest., 7-7 (). Harrill, J.A. et al. Transcriptional Response of Rat Frontal Cortex Following Acute in Vivo Exposure to the Pyrethroid Insecticides Permethrin and Deltamethrin. BMC Genomics 9, 6 (). Henn, A., Kirner, S. & Leist, M. TLr Hypersensitivity of Astrocytes as Functional Consequence of Previous Inflammatory Episodes. J. Immunol. 6, (). Hu, Y. et al. Netrin- Promotes Glioblastoma Cell Proliferation through Integrin B Signaling. Neoplasia, 9-7 (). Hutson, T.H., Buchser, W.J., Bixby, J.L., Lemmon, V.P. & Moon, L.D.F. Optimization of a 96-Well Electroporation Assay for Postnatal Rat Cns Neurons Suitable for Cost Effective Medium-Throughput Screening of Genes That Promote Outgrowth. Front. Mol. Neurosci. (). Ilyin, S.E. et al. Integrated Expressional Analysis: Application to the Drug Discovery Process. Methods 37, - (). Jain, S., van Kesteren, R.E. & Heutink, P. High Content Screening in Neurodegenerative Diseases. J. Vis. Exp. (). Jepson, S. et al. Lingo-, a Transmembrane Signaling Protein, Inhibits Oligodendrocyte Differentiation and Myelination through Intercellular Self-Interactions. J. Biol. Chem. 7, -9 (). Jossin, Y. & Cooper, J.A. Reelin, Rap and N-Cadherin Orient the Migration of Multipolar Neurons in the Developing Neocortex. Nat. Neurosci., (). Kaltenbach, L.S. et al. Composite Primary Neuronal High-Content Screening Assay for Huntington s Disease Incorporating Non-Cell-Autonomous Interactions. J. Biomol. Screen., 6-9 (). Kerrison, J.B., Duh, E.J., Yu, Y., Otteson, D.C. & Zack, D.J. A System for Inducible Gene Expression in Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 6, (). Kino, Y. et al. Intracellular Localization and Splicing Regulation of Fus/Tls Are Variably Affected by Amyotrophic Lateral Sclerosis-Linked Mutations. Nucleic Acids Res. 39, 7-79 (). Kuai, L. et al. Aak Identified as an Inhibitor of Neuregulin-/Erbb-Dependent Neurotrophic Factor Signaling Using Integrative Chemical Genomics and Proteomics. Chem. Biol., 9-96 (). Kuai, L. et al. Chemical Genetics Identifies Small-Molecule Modulators of Neuritogenesis Involving Neuregulin-/ Erbb Signaling. ACS Chem. Neurosci., 3-3 (). Kunzevitzky, N.J. et al. Foxn Is Required for Retinal Ganglion Cell Distal Axon Patterning. Mol. Cell. Neurosci. 6, 73-7 (). Larsson, D.E., Hassan, S.B., Oberg, K. & Granberg, D. The Cytotoxic Effect of Emetine and Cgp-7A Studied with the Hollow Fiber Model and Arrayscan Assay in Neuroendocrine Tumors in Vitro. Anticancer Agents Med. Chem., ().

9 Larsson, D.E., Wickstrom, M., Hassan, S., Oberg, K. & Granberg, D. The Cytotoxic Agents Nsc-9397, Brefeldin a, Bortezomib and Sanguinarine Induce Apoptosis in Neuroendocrine Tumors in Vitro. Anticancer Res. 3, 9-6 (). Le, M.T. et al. Microrna-b Is a Novel Negative Regulator of p3. Genes Dev. 3, 6-76 (9). Le, M.T.N. et al. Microrna-b Promotes Neuronal Differentiation in Human Cells by Repressing Multiple Targets. Mol. Cell. Biol. 9, 9-3 (9). Lerch, J.K. et al. Isoform Diversity and Regulation in Peripheral and Central Neurons Revealed through Rna-Seq. PLoS One 7 (). Leyva, M.J. et al. Identification and Evaluation of Novel Small Molecule Pan-Caspase Inhibitors in Huntington s Disease Models. Chem. Biol. 7, 9- (). Lie, M., Grover, M. & Whitlon, D.S. Accelerated Growth from Spiral Ganglion Neurons Exposed to the Rho Kinase Inhibitor H-. Neuroscience 69, -6 (). Lipinski, M.M. et al. Genome-Wide Analysis Reveals Mechanisms Modulating Autophagy in Normal Brain Aging and in Alzheimer s Disease. Proc. Natl. Acad. Sci. U. S. A. 7, 6-69 (). Liu, D. et al. Screening of Immunophilin Ligands by Quantitative Analysis of Expression and Outgrowth in Cultured Neurons and Cells. J. Neurosci. Methods 63, 3-3 (7). Liu, R. et al. Pinocembrin Protects against B-Amyloid-Induced Toxicity in Neurons through Inhibiting Receptor for Advanced Glycation End Products (Rage)-Independent Signaling Pathways and Regulating Mitochondrion- Mediated Apoptosis. BMC Med., (). Lodge, A.P., Langmead, C.J., Daniel, G., Anderson, G.W. & Werry, T.D. Performance of Mouse Neural Stem Cells as a Screening Reagent: Characterization of Pac Activity in Medium-Throughput Functional Assays. J. Biomol. Screen., 9-6 (). Lotharius, J. et al. Progressive Degeneration of Human Mesencephalic Neuron-Derived Cells Triggered by Dopamine-Dependent Oxidative Stress Is Dependent on the Mixed-Lineage Kinase Pathway. J. Neurosci., (). Low, J. et al. Knockdown of Cancer Testis Antigens Modulates Neural Stem Cell Marker Expression in Glioblastoma Tumor Stem Cells. J. Biomol. Screen., 3-39 (). MacGillavry, H.D. et al. Nfil3 and Camp Response Element-Binding Protein Form a Transcriptional Feedforward Loop That Controls Neuronal Regeneration-Associated Gene Expression. J. Neurosci. 9, - (9). Mallon, R. et al. Antitumor Efficacy Profile of Pki-, a Dual Phosphatidylinositol 3-Kinase/Mammalian Target of Rapamycin Inhibitor. Mol. Cancer Ther. 9, (). Matsui, H. et al. Pink and Parkin Complementarily Protect Dopaminergic Neurons in Vertebrates. Hum. Mol. Genet., 3-3 (3). McNeish, J. et al. High-Throughput Screening in Embryonic Stem Cell-Derived Neurons Identifies Potentiators of α-amino-3-hydroxyl--methyl--isoxazolepropionate-type Glutamate Receptors. J. Biol. Chem., (). Moore, D.L. et al. Klf Family Members Regulate Intrinsic Axon Regeneration Ability. Science 36, 9-3 (9).

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