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.
pET30a(+)-PE2
By onPlasmid: This resource can be used to express and purify PE2 protein for RNP delivery.
pU6-pegRNA-LRRK2-G2019S-3d
By onIt can be used to introduce LRRK2-G2019S mutation using prime editing.
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.
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.
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.
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.
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.
Brain slice physiology and optogenetics
By onThe protocol outlines standard external solutions for electrophysiology experiments.
Sst+ GPi output neurons provide direct feedback to key nodes of the basal ganglia and drive behavioral flexibility
By onHere, the authors propose that GPi Sst+ neurons drive behavioral flexibility through a direct feedback signal to balance the activity of key nodes in the basal ganglia.
Native gel lipid binding assay
By onThis protocol details how to perform a native gel lipid binding assay to evaluate the number of lipids bound per molecule of 3xFLAG-SHIP164Δ901-1099
Single-cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, aging, and disease
By onThis study used two complementary approaches in a Parkinson's disease mouse model and revealed insights into dopaminergic neuron characteristics, aging markers, and potential disease genes like CASR.
Co-immunoprecipitation using GFP-trap
By onProtocol describing the procedure to perform co-immunoprecipitation experiments in HEK293 cells using GFP-trap beads
Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy
By onA protocol where Golgi, isolated from cells by GolgiTAG immunoprecipitation (IP) can be prepared and imaged using TEM. This protocol can also be used to image any organelles isolated using various organelle-IP protocols that are available.
pCMV-pOTC-3H-GFP
By onExpression of GFP fused with pOTC-3H, a leader peptide from mitochondrial matrix enzyme ornithine transcarbamylase and a thermostable three helix-bundle
H9 ES AAVS1-NGN2 FAM134B-/-; PiggyBac-Keima-RAMP4
By onES cells modified to lack FAM134B and express Keima-RAMP4 ER-phagy reporter using CRISPR/Cas9. NEUROG2 construct added to AAVS1 locus. Transfected with SERP1 and mKeima. Derived from human embryonic stem cells at blastocyst stage.