Sample Collection and Measurement of Serum Neurofilament Light (NfL)
By onThis protocol details the steps for the preparation of serum from human blood samples and the measurement of NfL concentration using the NF-Light Advantage kit on the HD-X Analyzer (Quanterix, Billerica, MA).
Expression and purification GST-tagged ATG13:ATG101 constructs
By onExpression and purification of GST-tagged ATG13/ATG101 constructs.
pCAG-TSF-ATG13 (363-517)
By onPlasmid: Mammalian expression of TwinStrep-Flag tagged ATG13 (363-517).
Pseudogenes limit the identification of novel common transcripts generated by their parent genes
By onGenomic sequences with high sequence similarity cause short sequencing reads to align to multiple locations, thus complicating genomic analyses. The authors investigated the impact of pseudogenes on transcriptomic analyses.
AAV injection in the nodose ganglia of mouse
By onThe gut-brain axis links the visceral organs to the medulla oblongata via the vagus nerve. Accessing to the afferent vagal pathway is important to dissect the role of cell populations in the bidirectional communication between brain and body. The jugular-nodose ganglion (JNG) complex contains the cell bodies of many heterogenous neural subpopulations responsible for sensing the physiological and pathological conditions of the thoracic and abdominal organs. However, the study of these ganglia is challenging in small animals due to size and location. Hence, in this protocol we describe a practical surgical approach to the vagal trunk, and the JNG complex in mice.
Incentivized Vigor Task
By onCode for an fMRI task in which human participants use a joystick to make speeded reaches to cued targets. This task examines incentive effects on movement, part of the Socal Kinesia and Incentivization for Parkinson's Disease (SKIP) dataset.
pCAG-mcherry- WIPI2dC93E-cs- TEV -STREP
By onPlasmid for mammalian expression of human WIPI2d C93E with N-terminal mCherry and C-terminal Strep.
Single-cell dissociation of brain organoids
By onThis protocol details single cell dissociation of brain organoids. This protocol is part of a Collection of protocols (dx.doi.org/10.17504/protocols.io.8epv593dng1b/v1) for the paper "Glucocerebrosidase, a Parkinson´s disease-associated protein, is imported into mitochondria and regulates complex I assembly and function" (https://doi.org/10.21203/rs.3.rs-1521848/v1)
RNA extraction and quantitative PCR to assay inflammatory gene expression
By onProtocol for RT-qPCR to test levels of NF-kB response genes in a cell model transiently over-expressing Parkin. We found upregulation of key pro-inflammatory genes normalized to housekeeping gene, Gapdh.
Open Field Test to assess motor coordination in a mouse parkinsonian model
By onThe Open Field task is a simple sensorimotor test used to determine general activity levels, gross locomotor activity, and exploration habits in rodent models of CNS disorders. Open Field Test
Detection of Tau ubiquitylation
By onThis protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman Körner, Hauke Holthusen, Victoria A. Trinkaus, Irina Dudanova, Rubén Fernández-Busnadiego, Wolfgang Baumeister, David W. Sanders, Saurabh Gautam, Marc I. Diamond, F. Ulrich Hartl, Mark S. Hipp bioRxiv 2022.02.18.481043; doi: https://doi.org/10.1101/2022.02.18.481043
Rapalog-induced chemical dimerization experiments
By onThis protocol describes how to conduct Rapalog-induced chemical dimerization experiments.
cDNA clones for expression of MitoTag (OMP25) in mammalian cells
By onPlasmids for expression of OMP25 (MitoTag) in human and mouse cells.
Bulk NGS/allele quantification – Highly efficient generation of isogenic pluripotent stem cell models using prime editing
By onThis file contains sequencing results for amplicons covering specific regions of interest related to prime editing of hPSCs
pCAG-MBP-Foldon-ATG9 (830-839)
By onPlasmid: To make ATG9 C-terminal tail trimer for expression in mammalian cells.
immunofluorescent staining with anti-GFP and anti-CD63 antibodies
By onimmunofluorescent staining with anti-GFP and anti-CD63 antibodies
Fixing hippo neurons to assess endogenous NEMO during oxidative stress
By onProtocol describing the procedure for fixing Hippocampal rat neurons to assess endogenous NEMO during oxidative stress.
