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Current Protocols in Neuroscience最新文献

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Manipulating Gene Expression in Projection-Specific Neuronal Populations Using Combinatorial Viral Approaches 利用组合病毒方法操纵投射特异性神经元群体中的基因表达
Current Protocols in Neuroscience Pub Date : 2018-03-28 DOI: 10.1002/0471142301.ns0435s65
Bryan B. Gore, Marta E. Soden, Larry S. Zweifel
{"title":"Manipulating Gene Expression in Projection-Specific Neuronal Populations Using Combinatorial Viral Approaches","authors":"Bryan B. Gore,&nbsp;Marta E. Soden,&nbsp;Larry S. Zweifel","doi":"10.1002/0471142301.ns0435s65","DOIUrl":"10.1002/0471142301.ns0435s65","url":null,"abstract":"<p>The mammalian brain contains tremendous structural and genetic complexity that is vital for its function. The elucidation of gene expression profiles in the brain, coupled with the development of large-scale connectivity maps and emerging viral vector–based approaches for target-selective gene manipulation, now allows for detailed dissection of gene-circuit interfaces. This protocol details how to perform combinatorial viral injections to manipulate gene expression in subsets of neurons interconnecting two brain regions. This method uses stereotaxic injection of a retrograde transducing CAV2-Cre virus into one brain region, combined with injection of a locally transducing Cre-dependent AAV virus into another brain region. This technique is widely applicable to the genetic dissection of neural circuitry, as it enables selective expression of candidate genes, dominant-negatives, fluorescent reporters, or genetic tools within heterogeneous populations of neurons, based upon their projection targets. <i>Curr. Protoc. Neurosci</i>. 65:4.35.1-4.35.20. © 2013 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"65 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/0471142301.ns0435s65","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32841201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 60
A Guide to Creating and Testing New INTRSECT Constructs 创建和测试新的相交结构的指南
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.30
Lief E. Fenno, Joanna Mattis, Charu Ramakrishnan, Karl Deisseroth
{"title":"A Guide to Creating and Testing New INTRSECT Constructs","authors":"Lief E. Fenno,&nbsp;Joanna Mattis,&nbsp;Charu Ramakrishnan,&nbsp;Karl Deisseroth","doi":"10.1002/cpns.30","DOIUrl":"10.1002/cpns.30","url":null,"abstract":"<p>As the power of genetically encoded interventional and observational tools for neuroscience expands, the boundaries of experimental design are increasingly defined by limits in selectively expressing these tools in relevant cell types. Single-recombinase-dependent expression systems have been widely used as a means to restrict gene expression based on single features by combining recombinase-dependent viruses with recombinase-expressing transgenic animals. This protocol details how to create INTRSECT constructs and use multiple recombinases to achieve targeting of a desired gene to subsets of neurons that are defined by multiple genetic and/or topological features. This method includes the design and utilization of both viruses and transgenic animals: these tools are inherently flexible and modular and may be used in different combinations to achieve the desired gene expression pattern. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.30","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9339441","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 24
Generation and Characterization of Functional Human Hypothalamic Neurons 功能性人类下丘脑神经元的生成与表征
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.40
Peter Kirwan, Magdalena Jura, Florian T. Merkle
{"title":"Generation and Characterization of Functional Human Hypothalamic Neurons","authors":"Peter Kirwan,&nbsp;Magdalena Jura,&nbsp;Florian T. Merkle","doi":"10.1002/cpns.40","DOIUrl":"10.1002/cpns.40","url":null,"abstract":"<p>Neurons in the hypothalamus orchestrate homeostatic physiological processes and behaviors essential for life. Defects in the function of hypothalamic neurons cause a spectrum of human diseases, including obesity, infertility, growth defects, sleep disorders, social disorders, and stress disorders. These diseases have been studied in animal models such as mice, but the rarity and relative inaccessibility of mouse hypothalamic neurons and species-specific differences between mice and humans highlight the need for human cellular models of hypothalamic diseases. We and others have developed methods to differentiate human pluripotent stem cells (hPSCs) into hypothalamic neurons and related cell types, such as astrocytes. This protocol builds on published studies by providing detailed step-by-step instructions for neuronal differentiation, quality control, long-term neuronal maintenance, and the functional interrogation of hypothalamic cells by calcium imaging. Together, these protocols should enable any group with appropriate facilities to generate and study human hypothalamic cells. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"81 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.40","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39983414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 20
Animal Model of Chronic Migraine-Associated Pain 慢性偏头痛相关疼痛的动物模型
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.33
Laura S. Moye, Amynah A. A. Pradhan
{"title":"Animal Model of Chronic Migraine-Associated Pain","authors":"Laura S. Moye,&nbsp;Amynah A. A. Pradhan","doi":"10.1002/cpns.33","DOIUrl":"10.1002/cpns.33","url":null,"abstract":"<p>Migraine is a debilitating condition that affects hundreds of millions of people worldwide. A subset of these patients experience chronic migraine, resulting in long-term disability and a severely lowered quality of life. The development of novel migraine therapies has been slow, partially due to the small number of predictive animal models. We have recently developed a novel model of chronic migraine-associated pain, using the known human migraine trigger, nitroglycerin. Injection of nitroglycerin evokes an acute mechanical hyperalgesia, which is sensitive to the acute migraine therapy sumatriptan. In addition, chronic administration of nitroglycerin produces a progressive and sustained decrease in basal mechanical responses, and this hypersensitivity is blocked by migraine preventatives such as topiramate. This mouse model of chronic migraine can be used to study the mechanisms underlying progression of migraine from an episodic to a chronic disorder, and for identifying and screening novel acute and preventive migraine therapies. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.33","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35143524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 33
Time-Lapse Imaging and Cell Tracking of Migrating Cells in Slices and Flattened Telencephalic Vesicles 切片和扁平脑端小泡中迁移细胞的延时成像和细胞跟踪
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.24
Verónica Murcia-Belmonte, Giovanna Expósito, Eloísa Herrera
{"title":"Time-Lapse Imaging and Cell Tracking of Migrating Cells in Slices and Flattened Telencephalic Vesicles","authors":"Verónica Murcia-Belmonte,&nbsp;Giovanna Expósito,&nbsp;Eloísa Herrera","doi":"10.1002/cpns.24","DOIUrl":"10.1002/cpns.24","url":null,"abstract":"<p>Neuronal migration is a vital process needed for subsequent assembly and function of neural circuitry during embryonic development. The vast majority of neural progenitors are generated far from their final destination and need to migrate considerable distances to reach their specific cortical layer. Innovations in cell culture techniques and fluorescence microscopy now facilitate the direct visualization of cell movements during cortical development. Here, a detailed protocol to record and analyze a particular type of early migrating neurons, the Cajal Retzius Cells, during the development of the telencephalic vesicles in mammals is described. This method applied to other reporter mouse lines or to electroporated mouse embryos can be also used to analyze the migration of different types of moving neurons during cortical development. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"79 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.24","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34904457","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Cannabinoid-Induced Tetrad in Mice 大麻素诱导小鼠四分体
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.31
Mathilde Metna-Laurent, Miguel Mondésir, Agnès Grel, Monique Vallée, Pier-Vincenzo Piazza
{"title":"Cannabinoid-Induced Tetrad in Mice","authors":"Mathilde Metna-Laurent,&nbsp;Miguel Mondésir,&nbsp;Agnès Grel,&nbsp;Monique Vallée,&nbsp;Pier-Vincenzo Piazza","doi":"10.1002/cpns.31","DOIUrl":"10.1002/cpns.31","url":null,"abstract":"<p>Cannabinoid-induced tetrad is a preclinical model commonly used to evaluate if a pharmacological compound is an agonist of the central type-1 cannabinoid (CB1) receptor in rodents. The tetrad is characterized by hypolocomotion, hypothermia, catalepsy, and analgesia, four phenotypes that are induced by acute administration of CB1 agonists exemplified by the prototypic cannabinoid delta-9-tetrahydrocannabinol (THC). This unit describes a standard protocol in mice to induce tetrad phenotypes with THC as reference cannabinoid. We provide typical results obtained with this procedure showing a dose effect of THC in different mouse strains. The effect of the CB1 antagonist rimonabant is also shown. This tetrad protocol is well adapted to reveal new compounds acting on CB1 receptors in vivo. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.31","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35144602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 54
WGA-Alexa Conjugates for Axonal Tracing 用于轴突跟踪的WGA-Alexa共轭物
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.28
Sabrina L. Levy, Joshua J. White, Elizabeth P. Lackey, Lindsey Schwartz, Roy V. Sillitoe
{"title":"WGA-Alexa Conjugates for Axonal Tracing","authors":"Sabrina L. Levy,&nbsp;Joshua J. White,&nbsp;Elizabeth P. Lackey,&nbsp;Lindsey Schwartz,&nbsp;Roy V. Sillitoe","doi":"10.1002/cpns.28","DOIUrl":"10.1002/cpns.28","url":null,"abstract":"<p>Anatomical labeling approaches are essential for understanding brain organization. Among these approaches are various methods of performing tract tracing. However, a major hurdle to overcome when marking neurons in vivo is visibility. Poor visibility makes it challenging to image a desired neuronal pathway so that it can be easily differentiated from a closely neighboring pathway. As a result, it becomes impossible to analyze individual projections or their connections. The tracer that is chosen for a given purpose has a major influence on the quality of the tracing. Here, we describe the wheat germ agglutinin (WGA) tracer conjugated to Alexa fluorophores for reliable high-resolution tracing of central nervous system projections. Using the mouse cerebellum as a model system, we implement WGA-Alexa tracing for marking and mapping neural circuits that control motor function. We also show its utility for marking localized regions of the cerebellum after performing single-unit extracellular recordings in vivo. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"79 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.28","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34904458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 15
Multisite Electrophysiology Recordings in Mice to Study Cross-Regional Communication During Anxiety 小鼠多位点电生理记录研究焦虑时的跨区域交流
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.32
Alexander Z. Harris, Danielle Golder, Ekaterina Likhtik
{"title":"Multisite Electrophysiology Recordings in Mice to Study Cross-Regional Communication During Anxiety","authors":"Alexander Z. Harris,&nbsp;Danielle Golder,&nbsp;Ekaterina Likhtik","doi":"10.1002/cpns.32","DOIUrl":"10.1002/cpns.32","url":null,"abstract":"<p>Recording neural activity in awake, freely moving mice is a powerful and flexible technique for dissecting the neural circuit mechanisms underlying pathological behavior. This unit describes protocols for designing a drive and recording single neurons and local field potentials during anxiety-related paradigms. We also include protocols for integrating pharmacologic and optogenetic means for circuit manipulations, which, when combined with electrophysiological recordings, demonstrate input-specific and cell-specific contributions to circuit-wide activity. We discuss the planning, execution, and troubleshooting of physiology experiments during anxiety-like behavior. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.32","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35143525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 11
Fluorescence Microscopy: A Concise Guide to Current Imaging Methods 荧光显微镜:当前成像方法的简明指南
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.29
Christian A. Combs, Hari Shroff
{"title":"Fluorescence Microscopy: A Concise Guide to Current Imaging Methods","authors":"Christian A. Combs,&nbsp;Hari Shroff","doi":"10.1002/cpns.29","DOIUrl":"10.1002/cpns.29","url":null,"abstract":"<p>The field of fluorescence microscopy is rapidly growing and offers ever more imaging capabilities for biologists. Over the past decade, many new technologies and techniques have been developed that allow for combinations of deeper, faster, and higher resolution imaging. These have included the commercialization of many super-resolution and light sheet fluorescence microscopy techniques. For the non-expert, it can be difficult to match the best imaging techniques to biological questions. Picking the most appropriate imaging modality requires a basic understanding of the underlying physics governing each of them, as well as information comparing potentially competing imaging properties in the context of the sample to be imaged. To address these issues, we provide here concise descriptions of a wide range of commercially available imaging techniques from wide-field to super-resolution microscopy, and provide a tabular guide to aid in comparisons among them. In this manner we provide a concise guide to understanding and matching the correct imaging modality to meet research needs. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"79 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.29","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34904456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 158
Controlled Cortical Impact in the Rat 大鼠的控制性皮质冲击
Current Protocols in Neuroscience Pub Date : 2018-02-13 DOI: 10.1002/cpns.37
Dana D. Dean, Joseph A. Frank, L. Christine Turtzo
{"title":"Controlled Cortical Impact in the Rat","authors":"Dana D. Dean,&nbsp;Joseph A. Frank,&nbsp;L. Christine Turtzo","doi":"10.1002/cpns.37","DOIUrl":"10.1002/cpns.37","url":null,"abstract":"<p>Traumatic brain injury (TBI) is a major cause of death and disability world-wide. Following initial injury, TBI patients can face long-term disability in the form of cognitive, physical, and psychological deficits, depending on the severity and location of injury. This results in an economic burden in the United States estimated to be $60 billion due to health-care costs and loss of productivity. TBI is a significant area of active research interest for both military and civilian medicine. Numerous pre-clinical animal models of TBI are used to characterize the anatomical and physiological pathways involved and to evaluate therapeutic interventions. Due to its flexibility and scalability, controlled cortical impact (CCI) is one of the most commonly used preclinical TBI models. This unit provides a basic CCI protocol performed in the rat. © 2017 by John Wiley &amp; Sons, Inc.</p>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":"81 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.37","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35537292","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 9
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