National Institute of Neurological Disorders and Stroke 4 articles published in JoVE 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. 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. Kelly3,4, 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. Neuroscience Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons Seth Villarreal1, Sung Hoon Lee1,2, 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 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.
