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National Institute of Neurological Disorders and Stroke

9 ARTICLES PUBLISHED IN JoVE

Biology

Methods for Patch Clamp Capacitance Recordings from the Calyx

Kenneth Paradiso¹, Wei Wu¹, Ling-Gang Wu¹

¹National Institute of Neurological Disorders and Stroke, National Institute of Health

We demonstrate the basic techniques for presynaptic patch clamp recording at the calyx of Held, a mammalian central nervous system nerve terminal.

Neuroscience

Ex vivo Culturing of Whole, Developing Drosophila Brains

Ranjini Prithviraj^1,2, Svetlana Trunova^1,2, Edward Giniger^1,2

¹National Institute of Neurological Disorders and Stroke, ²National Human Genome Research Institute, National Institutes of Health, Bethesda, MD

This article describes a method by which one can mimic in vivo development of the Drosophila mushroom body in an ex vivo culture system.

Neuroscience

Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning

Nicholas J Luciano¹, Pascal Sati¹, Govind Nair¹, Joseph R Guy¹, Seung-Kwon Ha¹, Martina Absinta¹, Wen-Yang Chiang², Emily C Leibovitch³, Steven Jacobson³, Afonso C Silva², Daniel S. Reich¹

¹Translational Neuroradiology Section, National Institute of Neurological Disorders and Stroke, ²Cerebral Microcirculation Section, National Institute of Neurological Disorders and Stroke, ³Viral Immunology Section, National Institute of Neurological Disorders and Stroke

The overall goal of this protocol is to accurately align magnetic resonance imaging (MRI) image volumes with histology sections via the creation of customized 3D-printed brain holders and slicer boxes.

Neuroscience

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons

Seth Villarreal¹, Sung Hoon Lee^1,2, Ling-Gang Wu¹

¹National Institute of Neurological Disorders and Stroke, ²College of Pharmacy, Chung-ang University

Synaptic vesicle endocytosis is detected by light microscopy of pHluorin fused with synaptic vesicle protein and by electron microscopy of vesicle uptake.

Neuroscience

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Kaya J.E. Matson¹, Anupama Sathyamurthy¹, Kory R. Johnson², Michael C. Kelly^3,4, Matthew W. Kelley³, Ariel J. Levine¹

¹Spinal Circuits and Plasticity Unit, National Institute of Neurological Disorders and Stroke, ²Bioinformatics Section, Information Technology Program, National Institute of Neurological Disorders and Stroke, ³Laboratory of Cochlear Development, National Institute on Deafness and Other Communication Disorders, ⁴Single Cell Analysis Facility, Frederick National Laboratory

Here, we present a protocol to rapidly isolate high-quality nuclei from the fresh or frozen tissue for downstream massively parallel RNA sequencing. We include detergent-mechanical and hypotonic-mechanical tissue disruption and cell lysis options, both of which can be used for isolation of nuclei.

Bioengineering

3D Analysis of Multi-cellular Responses to Chemoattractant Gradients

Tae-Yun Kang¹, David Ellison², Sung Hoon Lee¹, Andrew J. Ewald^2,3, Andre Levchenko¹

¹Department of Biomedical Engineering and Yale Systems Biology Institute, Yale University, ²Department of Biomedical Engineering, Johns Hopkins University, ³Center for Cell Dynamics and Department of Cell Biology, Johns Hopkins University

We describe a method to construct devices for 3D culture and experimentation with cells and multicellular organoids. This device allows analysis of cellular responses to soluble signals in 3D microenvironments with defined chemoattractant gradients. Organoids are better than single cells at detection of weak noisy inputs.

Medicine

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Fuliang Luo¹, Xin Wang¹, Xiaokang Luo¹, Bo Li¹, Desheng Zhu², Hansong Sun³, Yue Tang¹

¹Beijing Key Laboratory of Pre-clinical Research and Evaluation for Cardiovascular Implant Materials, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, ²Laboratory Animal Center, Peking University, ³Center of Cardiac Surgery for Adults, State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Centre for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

Medicine

Technical Modification of the Terminal Ureter During Total Transperitoneal Laparoscopic Nephroureterectomy for Upper Urinary Tract Urothelial Carcinoma

Chunlong Fu^*1,2, Yaqun Zhang^*1, Xin Wang¹, Shengcai Zhu¹, Huimin Hou¹, Shengjie Liu¹, Cheng Pang¹, Xinda Song¹, Jia Chen¹, Jianye Wang¹, Ming Liu^1,2

¹(Urology) Beijing Hospital, National Center of Gerontology, The Joint Unit of the Chinese Academy of Medical Sciences, ²Graduate School of Peking Union Medical College, Beijing Hospital

Here, we present a protocol to increase the surgical field of view and reduce the difficulty of total transperitoneal laparoscopic nephroureterectomy surgery by precutting the umbilical ligament before treating the terminal ureter.

Neuroscience

Confocal Microscopy to Measure Three Modes of Fusion Pore Dynamics in Adrenal Chromaffin Cells

Sue Han^*1, Xin Wang^*1, Nicholas Cordero¹, Ling-Gang Wu¹

¹National Institute of Neurological Disorders and Stroke

This protocol describes a confocal imaging technique to detect three fusion modes in bovine adrenal chromaffin cells. These fusion modes include 1) close-fusion (also called kiss-and-run), involving fusion pore opening and closure, 2) stay-fusion, involving fusion pore opening and maintaining the opened pore, and 3) shrink-fusion, involving fused vesicle shrinkage.