Mouse Stereotaxic Surgery
By onThis protocol describes the steps for performing stereotaxic surgery in mice. It is applicable to intracranial injections (e.g. virus, drug) and placement of implants (e.g. optical fibers, electrode arrays) into targeted regions of mouse brains.
Microsomal membrane isolation from cell culture
By onMicrosomal membrane isolation from cell culture
Validation of Genotyping Method for L444P Mice Ear-Clips.
By on|| Team Schapira || Authors Revi Shahar Golan, David ChauAbstractAim: the genotyping is used to identify if mice are heterozygote (hetero) or Wild-Type (WT), and the aim of the work is to validate the digestion method, and PCR program, the PCR primers, and the interpretation of the results. Associated with publication: doi: 10.1093/brain/awx221
Untargeted lipidomics analysis for Golgi immunopurification (Golgi-IP)
By onThis protocol provides details for analyzing GolgiIP lipidomics samples using liquid chromatography mass spectrometry (LC-MS) for nonpolar lipid profiling.
Expression and purification MBP-ATG9 Constructs
By onExpression and purification from HEK cells of ATG13, ATG101 and FOLDON-ATG9A proteins.
CRISPR/Cas9-Based Functional Genomics in Human Induced Pluripotent Stem Cell–Derived Models: Can “the Stars Align” for Neurodegenerative Diseases?
By onThe article discusses the use of CRISPR/Cas9 in studying human diseases using stem cell models, highlighting its potential for advancing functional genomics research.
The annotation of GBA1 has been concealed by its protein-coding pseudogene GBAP1
By onThe authors identify novel transcripts from both GBA1 and GBAP1, including protein-coding transcripts that are translated in vitro and detected in proteomic data, but that lack GCase activity.
PGK EGFP-LC3B
By onPlasmid for mammalian expression of fluorescently-tagged LC3B, marker for autophagic vesicles.
Expression and purification Twin-STREP-FLAG tagged ATG13:ATG101 constructs
By onExpression and purification Twin-STREP-FLAG tagged ATG13:ATG101 constructs
Confirming circRNA expression by qPCR
By onThis protocol delineates a qPCR method to confirm the expression of circRNAs extracted from brain samples.
Glucosylceramidase Beta (GBA) Genotyping
By onThis protocol details the steps for GBA genotyping. This protocol has been adapted from the PRoBaND Clinical Consortium (incorporating methods described by Neuman et al., 2009 and Stone et al., 2000) and has been used for all publications for PRoBaND / Tracking Parkinson's describing clinical data and outcomes with respect to GBA status
Standard Operating Procedure: Mouse Stereotaxic Intracranial Injection Surgery
By onSOP outlines procedures for mouse stereotaxic intracranial injection surgery.
RBG Motif Bridge-Like Lipid Transport Proteins: Structure, Functions, and Open Questions
By onThe life of eukaryotic cells requires the transport of lipids between membranes, which are separated by the aqueous environment of the cytosol. Vesicle-mediated traffic along the secretory and endocytic pathways and lipid transfer proteins (LTPs) cooperate in this transport. Until recently, known LTPs were shown to carry one or a few lipids at a time and were thought to mediate transport by shuttle-like mechanisms. Over the last few years, a new family of LTPs has been discovered that is defined by a repeating β-groove (RBG) rod-like structure with a hydrophobic channel running along their entire length. This structure and the localization of these proteins at membrane contact sites suggest a bridge-like mechanism of lipid transport. Mutations in some of these proteins result in neurodegenerative diseases. Here we review the known properties and well-established or putative physiological roles of these proteins, and we highlight the many questions that remain open about their functions.
Sex-specific microglial responses to glucocerebrosidase inhibition
By onMorpho-dynamic analysis occurring in primary cells derived from female and male mice in response to proinflammatory stimulations and glucocerebrosidase inhibition.
SoRA microscopy protocol for imaging oligomers in human brain tissue
By onThis protocol gives a step-by-step guide to imaging oligomers in human brain tissue using a spinning disk confocal microscope.
Flow cytometry-based measurement of trafficking of Golgi to the lysosome via autophagy
By onA protocol was developed to measure Golgi trafficking to lysosomes through autophagy in HEK293 cells using Keima-YIPF3/4 reporters with flow cytometry.