Current Protocols in Neuroscience最新文献_第5页

Current Protocols in Neuroscience Pub Date : 2019-03-28 DOI: 10.1002/cpns.60

{"title":"Issue Information TOC","authors":"","doi":"10.1002/cpns.60","DOIUrl":"10.1002/cpns.60","url":null,"abstract":"Cover: In Erben and Buonanno (https://doi.org/10.1002/cpns63), the image shows examples for chromogenic BaseScope detection (Basic Protocol 2) and ISH-IHC combination (Basic Protocol 3). (A) Hippocampal section of an adult WT mice, labeled using BaseScope for ErbB4 with a single probe pair targeting the exon boundary exon 1/exon 2. The section was counterstained with hematoxylin and signal detected with light microscopy. Arrowheads indicate ErbB4+ GABAergic interneurons. (B) Detection of one of the four ErbB4 splice variants (JMb; cyan) by BaseScope in GABAergic interneurons (GAD-GFP; green; arrowheads) in a section of an adult GAD-GFP mouse (kindly provided by Dr. Yuchio Yanagawa). GFP signal was amplified after the ISH assay with an anti-GFP antibody (NeuroMab; N86/8; RRID: AB_10671444). (C) ISH for ErbB4 (C1; cyan) and Th (C3; magenta) was combined with IHC with an antibody against DAT (green; Santa Cruz, sc-32258; RRID: AB_627400) in a sagittal FFPE section from an adultWT mouse; depicted substantia nigra compacta (SNc) on the left and dopaminergic medial forebrain bundle. (C′) Magnification of ErbB4-expressing dopaminergic neurons (arrowheads) in the SNc. (D) In primary mesencephalic cultures (DIV8; prepared as in Skirzewski et al., 2018), DAT (green) immunostaining was performed post-hoc to RNAscope for ErbB4 (C1; cyan) and Th (C3; magenta). Scale bars 100 µm in B, 20 µm in other panels. Abbreviations: ISH-IHC, in situ hybridization-immunohistochemistry; GFP, green fluorescent protein; FFPE, formalin-fixed paraffin-embedded; WT, wild type.\u0000\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture>\u0000 </div>\u0000 </figure>","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.60","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72550266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Recombinant Viral Vectors as Neuroscience Tools 重组病毒载体作为神经科学工具

Current Protocols in Neuroscience Pub Date : 2019-03-22 DOI: 10.1002/cpns.67

Shih-Heng Chen, Juhee Haam, Mitzie Walker, Erica Scappini, John Naughton, Negin P. Martin

引用次数: 23

Production of Viral Constructs for Neuroanatomy, Calcium Imaging, and Optogenetics 用于神经解剖学、钙成像和光遗传学的病毒构建体的生产

Current Protocols in Neuroscience Pub Date : 2019-03-18 DOI: 10.1002/cpns.66

Shih-Heng Chen, Juhee Haam, Mitzie Walker, Erica Scappini, John Naughton, Negin P. Martin

{"title":"Production of Viral Constructs for Neuroanatomy, Calcium Imaging, and Optogenetics","authors":"Shih-Heng Chen, Juhee Haam, Mitzie Walker, Erica Scappini, John Naughton, Negin P. Martin","doi":"10.1002/cpns.66","DOIUrl":"10.1002/cpns.66","url":null,"abstract":"Advances in design and use of light-sensitive and light-emitting sensors have facilitated observation, measurement, and control of neuronal activities. Viruses are effective vectors for delivery of these valuable research tools to mammalian brains. Recombinant viruses are optimized to mediate regulatable, long-term, and cell-specific gene expression. Here, we describe production methods for three of the most commonly used types of recombinant viruses in neurobiology research: adeno-associated virus (AAV), retrovirus/lentivirus, and glycoprotein-deleted rabies virus. These viral constructs are frequently used for calcium imaging or to deliver neural tracers and optogenetic tools. Popular constructs are readily obtained commercially; however, customized virus production through commercial sources is time consuming and costly. This article aims to provide readers with detailed technical information for rapid production and validation of high-quality viral particles in a laboratory setting while highlighting advantages and limitations of each viral type. © 2019 by John Wiley & Sons, Inc.","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.66","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37066214","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: 12

In Vivo Electroporation and Time-Lapse Imaging of the Rostral Migratory Stream in Developing Rodent Brain 发育中的啮齿动物大脑吻侧迁移流的体内电穿孔和延时成像

Current Protocols in Neuroscience Pub Date : 2019-03-12 DOI: 10.1002/cpns.65

Zhihui Huang, Ying Wang

引用次数: 2

Analysis of Neuro-Neuronal Synapses Using Embryonic Chick Ciliary Ganglion via Single-Axon Tracing, Electrophysiology, and Optogenetic Techniques 利用单轴突示踪、电生理和光遗传技术分析胚胎鸡睫状神经节的神经突触

Current Protocols in Neuroscience Pub Date : 2019-02-21 DOI: 10.1002/cpns.64

Ryo Egawa, Hiromu Yawo

引用次数: 1

Detection and Quantification of Multiple RNA Sequences Using Emerging Ultrasensitive Fluorescent In Situ Hybridization Techniques 使用新兴的超灵敏荧光原位杂交技术检测和定量多个RNA序列

Current Protocols in Neuroscience Pub Date : 2019-02-21 DOI: 10.1002/cpns.63

Larissa Erben, Andres Buonanno

{"title":"Detection and Quantification of Multiple RNA Sequences Using Emerging Ultrasensitive Fluorescent In Situ Hybridization Techniques","authors":"Larissa Erben, Andres Buonanno","doi":"10.1002/cpns.63","DOIUrl":"10.1002/cpns.63","url":null,"abstract":"Fluorescent detection of transcripts using RNAscope has quickly become a standard in situ hybridization (ISH) approach in neuroscience with over 400 publications since its introduction in 2012. RNAscope's sensitivity and specificity allow the simultaneously detection of up to three low abundance mRNAs in single cells (i.e., multiplexing) and, in contrast to other ISH techniques, RNAscope is performed in 1 day. BaseScope, a newer ultrasensitive platform, uses improved amplification chemistry of single oligonucleotide probe pairs (∼50 bases). This technique allows discrimination of single nucleotide polymorphisms or splice variants that differ by short exons. A present limitation of BaseScope is that expression analysis is limited to a single gene (i.e., single-plexing). This article outlines detailed protocols for both RNAscope and BaseScope in neuronal tissue. We discuss how to perform ISH experiments using either fresh-frozen or formalin-fixed paraffin-embedded sections, as well as dissociated cultured neurons. We also outline how to obtain quantitative data from hybridized tissue sections. © 2019 by John Wiley & Sons, Inc.","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2019-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.63","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37148390","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

Issue Information TOC 发布信息TOC

Current Protocols in Neuroscience Pub Date : 2019-01-03 DOI: 10.1002/cpns.59

引用次数: 0

High-Throughput Analysis of Behavior Under the Control of Optogenetics in Caenorhabditis elegans 秀丽隐杆线虫光遗传学控制下行为的高通量分析

Current Protocols in Neuroscience Pub Date : 2018-11-02 DOI: 10.1002/cpns.57

Alex J. Yu, Troy A. McDiarmid, Evan L. Ardiel, Catharine H. Rankin

引用次数: 8

Miniscope GRIN Lens System for Calcium Imaging of Neuronal Activity from Deep Brain Structures in Behaving Animals 微型GRIN透镜系统对行为动物脑深部结构的神经元活动进行钙成像

Current Protocols in Neuroscience Pub Date : 2018-10-13 DOI: 10.1002/cpns.56

Lifeng Zhang, Bo Liang, Giovanni Barbera, Sarah Hawes, Yan Zhang, Kyle Stump, Ira Baum, Yupeng Yang, Yun Li, Da-Ting Lin

{"title":"Miniscope GRIN Lens System for Calcium Imaging of Neuronal Activity from Deep Brain Structures in Behaving Animals","authors":"Lifeng Zhang, Bo Liang, Giovanni Barbera, Sarah Hawes, Yan Zhang, Kyle Stump, Ira Baum, Yupeng Yang, Yun Li, Da-Ting Lin","doi":"10.1002/cpns.56","DOIUrl":"10.1002/cpns.56","url":null,"abstract":"Visualizing neural activity from deep brain regions in freely behaving animals through miniature fluorescent microscope (miniscope) systems is becoming more important for understanding neural encoding mechanisms underlying cognitive functions. Here we present our custom-designed miniscope GRadient INdex (GRIN) lens system that enables simultaneously recording from hundreds of neurons for months. This article includes miniscope design, the surgical procedure for GRIN lens implantation, miniscope mounting on the head of a mouse, and data acquisition and analysis. First, a target brain region is labeled with virus expressing GCaMP6; second, a GRIN lens is implanted above the target brain region; third, following mouse surgical recovery, a miniscope is mounted on the head of the mouse above the GRIN lens; and finally, neural activity is recorded from the freely behaving mouse. This system can be applied to recording the same population of neurons longitudinally, enabling the elucidation of neural mechanisms underlying behavioral control. © 2018 by John Wiley & Sons, Inc.","PeriodicalId":40016,"journal":{"name":"Current Protocols in Neuroscience","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2018-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpns.56","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36571152","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: 63

Issue Information TOC 发布信息TOC

Current Protocols in Neuroscience Pub Date : 2018-10-08 DOI: 10.1002/cpns.58

引用次数: 0