Thawing, Passaging and Freezing of hPSCs on MEFs
By onThis collection contains protocols which describe the standard procedure of culturing human pluripotent stem cells (hPSCs) on inactivated mouse embryonic fibroblasts (MEFs).
RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of synaptic vesicle precursors
By onGain-of-function mutations in LRRK2 gene lead to by increasing phosphorylation of RAB GTPases. Over-phosphorylation of RAB3A disrupts synaptic vesicle transport in human neurons, altering synaptic protein distribution and likely contributing to PD.
The GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in human cell lines
By onThe GBA variant E326K is associated with alpha-synuclein aggregation and lipid droplet accumulation in fibroblasts.
CD8 Cell Density In Substantia Nigra And Cerebral Peduncle Image Analysi
By onQuPath is a bioimage analysis software designed for digital pathology and whole slide image analysis. This protocol describes how to measure CD8 density in the substantia nigra and cerebral peduncle using haematoxylin and DAB-stained brain sections.
DOPA pheomelanin is increased in nigral neuromelanin of Parkinson’s disease
By onNeuromelanin in the substantia nigra may be a key factor contributing to dopaminergic neuron vulnerability in Parkinson’s disease. Here, the authors investigated the relative composition and specific roles of pheomelanin and eumelanin in PD.
A prebiotic diet modulates microglia response and motor deficits in α-synuclein overexpressing mice
By onSingle-cell RNA-seq analysis of microglia from the substantia nigra and striatum uncovers increased pro-inflammatory signaling and reduced homeostatic responses in ASO mice compared to wild-type counterparts on standard diets. However, prebiotic feeding reverses pathogenic microglial states in ASO mice and promotes expansion of protective disease-associated macrophage (DAM) subsets of microglia. Notably, depletion of microglia using a CSF1R inhibitor eliminates the beneficial effects of prebiotics by restoring motor deficits to ASO mice despite feeding a prebiotic diet. These studies uncover a novel microglia-dependent interaction between diet and motor symptoms in mice, findings that may have implications for neuroinflammation and PD.
Preparing mRNA for nucleofection of hPSCs
By onThis protocol describes the standard procedure for preparing mRNA to be delivered into human pluripotent stem cells (hPSCs) using nucleofection.
Microscopy-based pUb-coverage measurements of mitochondria in iNeurons
By onAssociated with preprint:
Lentivirus plasmids for sgRNA: pLV[Exp]-U6>NT-Seq1-hPGK>mApple
By onPlasmid: Plasmid vector encoding a non-targeting sgRNA sequence under a U6 promoter and a mApple fluorescent reporter. Generated by Vectorbuilder in the pLV backbone
H9 ES AAVS1-NGN2 FLAG-EEA1
By onES cells were modified to create iNeurons expressing EEA1 with a 3xFLAG tag. NEUROG2 was introduced at the AAVS1 locus using CRISPR/Cas9. The cells are derived from human embryonic stem cells at the blastocyst stage.
H9 ES AAVS1-NGN2 FAM134C/A-/-
By onES cells modified using CRISPR/Cas9 to lack FAM134C & A ER-phagy receptor. NEUROG2 construct introduced in AAVS1 locus. Derived from human embryonic stem cells at blastocyst stage.
LiD genetic determinants study under CPH regression models
By onCode to perform the study of LiD genetic determinants under CPH regression models and functional annotation analyses.
Brain Slice Preparation for electrophysiology recording
By onThis protocol describes brain slice preparation for electrophysiology recording.
Datasets and Key Resources Table used in Do, Quyen B. et al. (2023) “Early striatal hyperexcitability in an in vitro human striatal microcircuit model carrying the Parkinson’s GBA-N370S mutation”
By onThe files found here are associated to Do, Q. et al (2023) study and contain: (a) all source and supplementary data, (b) Key Resources Table, and (c) list of primers.
CRISPR/Cas9 generation of knock-out iPSCs
By onThis protocol describes RNP-based CRISPR/Cas9 gene-editing to generate knock-out iPSCs.
Early deficits in an in vitro striatal microcircuit model carrying the Parkinson’s GBA-N370S mutation
By onThe results highlight the unique utility of modeling striatal neurons in a modular and highly physiological circuit, which is essential to reveal mechanistic insights of the loss of electrical functional integrity in the striata of GBA1 PD patients.
