Whole-cell patch-clamping of cultured human neurons
By onThis protocol describes procedures of the whole-cell patch clamping of human neurons derived from induced pluripotent stem cells and cultured in adherent monolayer.
Erythroid Differentiation Dependent Interaction of VPS13A with XK at the Plasma Membrane of K562 Cells
By onMutations in VPS13A and XK lead to Chorea Acanthocytosis and McLeod syndrome, causing neurodegeneration and abnormal red blood cells. VPS13A binds XK at the ER and plasma membrane contacts in differentiated erythroblasts.
Dopamine transporter and synaptic vesicle sorting defects underlie auxilin-associated Parkinson’s disease
By onAuxilin participates in the uncoating of clathrin-coated vesicles (CCVs), thereby facilitating synaptic vesicle (SV) regeneration at presynaptic sites. Auxilin (DNAJC6/PARK19) loss-of-function mutations cause early-onset Parkinson's disease (PD). Here, we utilized auxilin knockout (KO) mice to elucidate the mechanisms through which auxilin deficiency and clathrin-uncoating deficits lead to PD. Auxilin KO mice display cardinal features of PD, including progressive motor deficits, α-synuclein pathology, nigral dopaminergic loss, and neuroinflammation. Significantly, treatment with L-DOPA ameliorated motor deficits. Unbiased proteomic and neurochemical analyses of auxilin KO brains indicated dopamine dyshomeostasis. We validated these findings by demonstrating slower dopamine reuptake kinetics in vivo, an effect associated with dopamine transporter misrouting into axonal membrane deformities in the dorsal striatum. Defective SV protein sorting and elevated synaptic autophagy also contribute to ineffective dopamine sequestration and compartmentalization, ultimately leading to neurodegeneration. This study provides insights into how presynaptic endocytosis deficits lead to dopaminergic vulnerability and pathogenesis of PD.
Endogenous coimmunoprecipitation
By onProtocol used for immunoprecipitation of HSP60 and LONP1 in HEK cells to show the interaction with V5-Flag-tagged WT-GCase
Orchestration of selective autophagy by cargo receptors
By onThe authors review recent insights into the mechanisms of action of cargo receptors in selective autophagy by focusing on the roles of sequestosome-like cargo receptors in the degradation of misfolded, ubiquitinated proteins and damaged mitochondria.
Genome-wide screen reveals Rab12 GTPase as a critical activator of Parkinson’s disease-linked LRRK2 kinase
By onThe data support a model in which Rab12 binding to a new site in the LRRK2 Armadillo domain activates LRRK2 kinase for Rab phosphorylation
Gene editing of YIPF4, CALCOCO1, and ATG7 in HeLa cells
By onThe text outlines the process of generating YIPF4, FIP200, and ATG7 knockout cell lines in HeLa cells through CRISPR-Cas9 technology.
Microscopy-based evaluation of Parkin translocation and mitophagy in FBXO7-/- cell lines
By onAssociated with preprint:
Impact of 100 LRRK2 variants linked to Parkinson’s Disease on kinase activity and microtubule binding
By onThis study systematically investigates 100 LRRK2 variants linked to PD, and provides rationale for variant carrier inclusion/exclusion in ongoing and future LRRK2 inhibitor clinical trials.
Loss of tau expression attenuates neurodegeneration associated with α-synucleinopathy
By onThe authors examine the role of tau in the onset and progression of αS pathology using a transgenic mouse model of α-synucleinopathy lacking mouse tau expression.
Genome-wide Analysis of Motor Progression in Parkinson Disease
By onThe genetic basis of Parkinson disease (PD) motor progression is largely unknown. Previous studies of the genetics of PD progression have included small cohorts and shown a limited overlap with genetic PD risk factors from case-control studies. Here, we have studied genomic variation associated with PD motor severity and early-stage progression in large longitudinal cohorts to help to define the biology of PD progression and potential new drug targets.
pU6-pegRNA-HEK3-CTTins-px330-scaffold
By onPlasmid: This resource can be used to introduce HEK3 CTT insertion using prime editing. It serves as a good positive control for prime editing.
Transferrin uptake assay to measure Clathrin-mediated endocytosis in HIPCS derived neurons
By onThis protocol is used to measure Transferrin uptake as a way to investigate Clathrin-mediated endocytosis in HIPCS derived neurons, or other cell types.
Structure of human ULK1 complex core (2:2:2 stoichiometry) of the ATG13(450-517) mutant
By onStructure of human ULK1 complex core (2:2:2 stoichiometry) of the ATG13(450-517) mutant
Duplicating 96-well plate-cultured hPSCs clones
By onThis protocol describes a standard procedure for duplicating 96-well plate-cultured human pluripotent stem cells (hPSCs).
Primary data associated with doi: 10.1101/2022.04.25.489459 (A Feed-forward Pathway Drives LRRK2 kinase Membrane Recruitment and Apparent Activation)
By onRaw immunoblotting data; tabular data for quantifications; immunofluorescence images.
Code for making forest plots for top GWAS loci
By onThis repository contains the script used to create forest plots for top loci .
