Cell culture, transfection, and imaging
By onThis protocol describes general procedures for culturing HeLa cells, transient transfection, and imaging using an Andor Dragonfly spinning disk confocal system.
PTEN-induced kinase 1 (PINK1) and Parkin: Unlocking a mitochondrial quality control pathway linked to Parkinson’s disease
By onThis review focuses on understanding the PINK1/Parkin-mediated mitochondrial quality control pathway through the lens of abherrant immune activation as a driver of dopaminerigic neuron loss following the loss of PINK and Parkin.
Differentiation of human medium spiny neurons (MSNs) from induced pluripotent stem cells (iPSCs)
By onThis protocol generates human medium spiny neurons (MSNs) from induced human pluripotent stem cells (iPSCs).
Live-imaging of axonal cargoes in human iPSC-derived neurons or mouse primary neurons
By onThe authors describe the procedure and equipment used for live imaging of axonal cargoes.
HEK293::TMEM192-3xHA YIPF4-/-
By onHEK293 cell line with TMEM192 gene C-terminally tagged with 3xHA epitope using CRISPR/Cas9. YIPF4 knockout also done. Derived from fetal kidney, human origin, adenovirus transformed.
The lipid flippase ATP10B enables cellular lipid uptake under stress conditions
By onATP10B mutations are linked to Parkinson's and Lewy body disease. ATP10B acts as a lipid transporter in late endo-/lysosomes, enhancing phosphatidylcholine uptake in cells under stress conditions like rotenone treatment.
Nuclei Isolation and Sorting from Frozen Human Temporal Cortex V2
By onProtocol focuses on isolating and sorting nuclei from frozen human temporal cortex.
GI Transit Assay
By onThis test is used to check for possible signs of constipation and alteration in gut transit.
Pathogenic LRRK2 control of primary cilia and Hedgehog signaling in neurons and astrocytes of mouse brain
By onPathogenic mutations in LRRK2 can cause loss of primary cilia in neurons. The authors show that cilia loss is seen very early in mice harboring the most common LRRK2 mutation.
Behavioral Testing – Open Field and Dyskinesia Scoring
By onThis protocol describes behavioral testing to assess motor deficits in mice. It includes open field locomotor testing and scoring of levodopa-induced dyskinesia.
Perfusion-fixation procedure for adult rhesus monkeys
By onThe protocol details the perfusion-fixation procedure for rhesus monkeys
Primary data associated with the manuscript “Parkinson’s VPS35[D620N] mutation induces LRRK2 mediated lysosomal association of RILPL1 and TMEM55B” (doi: 10.1101/2023.06.07.544051)
By onConfocal microscopy images, CellProfiler Excel files of Pearson's coefficients between TMEM55B/RILPL1 or pRab10/RILPL1 in R1441C MEF or VPS35 MEF cells as seen in doi: 10.1101/2023.06.07.544051.
HeLa ::TMEM192-HA CALCOCO1-/-
By onHeLa ::TMEM192-HA CALCOCO1-/- PubMed=37757899; Population: African American. Characteristics: Using CRISPR/Cas9 TMEM192 was C-terminally tagged on both alleles with a 3xHA epitope (from parent cell line). Knockout cell: Method=CRISPR/Cas9; HGNC; 29306; CALCOCO1. Transformant: NCBI_TaxID; 333761; Human papillomavirus type 18 (HPV18). Derived from site: In situ; Uterus, cervix; UBERON=UBERON_0000002. NCIt; C27677; Human papillomavirus-related cervical adenocarcinoma NCBI_TaxID=9606; ! Homo sapiens (Human) RRID:CVCL_D1KR ! HeLa::TMEM192-3xHA Female 30Y6M Cancer cell line
Seeding nucleofected hPSCs in 96-well plates using limited dilution
By onThis protocol describes a standard procedure for seeding nucleofected human pluripotent stem cells in 96-well plates using limited dilution. This protocol follows nucleofection of hPSCs as described in detail in the collection "Nucleofection (Amaxa) and electroporation (Biorad) of hPSCs;" dx.doi.org/10.17504/protocols.io.b4qnqvve
Investigation_of_mitophagy_in_Hippo_neurons
By onWe developed a method for assessing mitochondrial clearance in primary hippocampal neurons.
