Lysosome proteolysis analysis with DQ-BSA
By onProtocol to analyze lysosomal proteolysis in astrocytes generated from iPSCs.
Immunohistochemistry of Human Brain Striatum: ciliation of cholinergic neurons and astrocytes
By onThe authors present a method to identify the same brain region in human postmortem tissue to evaluate ciliary status. The striatum is identified morphologically and also by immunofluorescence microscopy using anti-DARPP-32 antibodies.
GFP-YIPF3 Immunoprecipitation
By onThe protocol outlines the immunoprecipitation procedure for GFP-YIPF3 protein.
Analysis of glycosphingolipids from human cerebrospinal fluid
By onThe method uses the fluorescent compound anthranilic acid (2-AA) to label oligosaccharides prior to analysis using normal-phase high-performance liquid chromatography. The labeling procedure is rapid, selective, and easy to perform.
Gene editing of YIPF4, FIP200, and ATG7 in HEK293 cells
By onProtocol outlines creation of YIPF4, FIP200, and ATG7 knockout cell lines in HEK293 cells via CRISPR-Cas9 method.
pFastBac_Dual_GST-TEV-EGFP-TBK1
By onPlasmid: Plasmid for the expression and purification of GST-TEV-EGFP-TBK1 in Spodoptera frugiperda cells (Sf9). Internal reference: SMC1631
From structure to ætiology: a new window on the biology of leucine-rich repeat kinase 2 and Parkinson’s disease
By onThis review summarizes LRRK2 structure both in a historical and current context, highlighting new insights into the structure of LRRK2 and complexes it forms.
pU6-pegRNA-LRRK2-G2019S-3d
By onIt can be used to introduce LRRK2-G2019S mutation using prime editing.
pET30a(+)-PE2
By onPlasmid: This resource can be used to express and purify PE2 protein for RNP delivery.
Determination of edits in CRISPR-edited cell lines by sequencing
By onThis protocol details the procedure of determination of edits in CRISPR-edited cell lines by sequencing.
Formalin Fixed Paraffin Embedded (FFPE) Tissue Preparation for Digital Pathology Quantification Using QuPath
By onQuPath is an open-source digital pathology analysis software. Here, it is used to quantify various misfolded proteins across anatomical regions.
Subcellular and regional localization of mRNA translation in midbrain dopamine neurons
By onLocal translation within excitatory and inhibitory neurons is known to be involved in neuronal development and synaptic plasticity. Despite the extensive dendritic and axonal arborizations of monoaminergic neurons, the subcellular localization of protein synthesis has not been well-characterized in these populations. Here, we investigated mRNA localization in midbrain dopaminergic (mDA) neurons, cells with enormous axonal and dendritic projections, both of which can release dopamine (DA). Using highly-sensitive sequencing and imaging approaches in mDA axons, we found no evidence for axonal mRNA localization or translation. In contrast, we found that mDA neuronal dendritic projections into the substantia nigra reticulata (SNr) contain ribosomes and mRNAs encoding the DA synthesis, release, and reuptake machinery. Surprisingly, we found dendritic localization of mRNAs encoding synaptic vesicular release proteins in mDA neurons. Our results are consistent with a role for local translation in the regulation of DA transmission from dendrites, but not striatal axons. Finally, we defined a molecular signature of sparse mDA neurons in the SNr, including enrichment of an ER calcium pump previously undescribed in mDA neurons. RNA-sequencing of brain tissue, RiboTagged dopamine neurons, and synaptosomes using PLATE-seq 3'-tag sequencing or Clontech SMARTer full-length RNA library preparation This dataset is a part of "Subcellular and regional localization of mRNA translation in midbrain dopamine neurons" (Hobson et. al, 2022).
Neuronal Presentation of Antigen and Its Possible Role in Parkinson’s Disease
By onPatients with Parkinson's disease and synucleinopathies show autoimmune features, with T cells recognizing alpha-synuclein. Studies explore T cell-mediated neuronal death in PD and other disorders.
Expansion microscopy with R1441C LRRK2 MEF cells: visualization of Myc-RILPL1 and TMEM55B
By onExpansion microscopy is a super-resolution imaging technique that increases the physical distance between fluorophores from fixed cells on coverslips. Here, the authors use expansion techniques to confocal image TMEM55B and RILPL1.
Harvesting and irradiation of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By onThis protocol describes the process of harvesting and irradiating mouse embryonic fibroblasts (MEFs) to use as feeder cells for human pluripotent stem cell (hPSC) culture.