Chemical degradation and determination of pheomelanin and eumelanin markers
By Emma Sherrell onThis is a protocol involving chemical oxidation and reduction methods followed by high-performance liquid chromatography (HPLC) detection of markers for pheomelanin and eumelanin.
Proteome preparation and analysis
By Julia Julia Leonard onProtocol for proteomic analysis from mouse cecal samples.
3′ mRNA-sequencing
By Julia Julia Leonard onProtocol for 3' mRNA sequencing of mouse gastrointestinal tract tissues.
Whole gut transit time, fecal water content, and fecal output
By Julia Julia Leonard onProtocol for mouse gastrointestinal function assays (whole gut transit time, fecal water content, and fecal output).
Characterization of splay characteristics of carbon fiber electrode threads (CFETs) in agar brain phantom
By Emma Sherrell onMethods to measure deflection characteristics of carbon fiber electrodes in agar brain phantoms.
Colonic migrating motor complexes
By Julia Julia Leonard onProtocol for ex vivo assay of colonic migrating motor complexes (CMMCs) in mouse colons.
Central and peripheral innate and adaptive immunity in Parkinson’s disease
By Emma Sherrell onParkinson’s disease (PD) is a multisystem disorder with characteristics of a chronic inflammatory disease. To develop effective immunomodulatory interventions to combat PD, researchers need to think innovatively about the implications of orchestrated central and peripheral innate and adaptive immune responses that occur as the disease begins and progresses.
CD8 cell density In substantia nigra and cerebral peduncle image analysis
By Emma Sherrell 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.
Pathways controlling neurotoxicity and proteostasis in mitochondrial complex I deficiency
By Julia Julia Leonard onNeuromuscular disorders caused by dysfunction of the mitochondrial respiratory chain are common, severe, and untreatable. The authors recovered a number of mitochondrial genes, including electron transport chain components, in a large forward genetic screen for mutations causing age-related neurodegeneration in the context of proteostasis dysfunction.
Cell viability assessment
By Emma Sherrell onThis is the protocol for assessing cell viability using a MTS cell proliferation assay kit.
Evaluating GPNMB ACD mutants by Western Blotting and immunofluorescence
By Julia Julia Leonard onThis protocol describes the evaluation of cellular processing of GPNMB mutants by Western Blotting and Immunofluorescent imaging in a HEK293 overexpression model.
Mitochondrial antigen presentation in RAW macrophages
By Emma Sherrell onThis protocol details methods for 3-day Mitochondrial Antigen Presentation Assay in a murine macrophage cell line (RAW) that expresses a glycoprotein B (gB) of herpes simplex virus 1 (HSV1) targeted to the mitochondrial matrix (mito-gB). Watch an interview about this protocol with Ahmed Fahmy, PhD.
Loss of primary cilia and dopaminergic neuroprotection in pathogenic LRRK2-driven and idiopathic Parkinson’s disease
By Julia Julia Leonard onPublished: Activating LRRK2 mutations causes Parkinson's disease. Previously, work showed that cholinergic interneurons and astrocytes lose primary cilia in LRRK2 mutant mice. Here, the data strongly suggest that loss of cilia in specific striatal cell types decreases neuroprotection for dopamine neurons in mice and human Parkinson's disease. View original preprint.
Neural differentiation on EM grids – iNeurons sample preparation for cryo-ET and CLEM
By Emma Sherrell onA protocol for differentiating AAVS1-TRE3G-NGN2 hESCs to iNeurons directly on EM grids for cryo-ET and cryo-CLEM. The protocol includes: (1) EM grids coating with Matrigel or Poly-Ornithine and Laminin; (2) differentiation and seeding of iNeurons on EM grids; (3) lentiviral transduction for transient expression of fluorescently-labeled proteins; and (4) plunge freezing. Watch an interview about this protocol with Cristina Capitanio, MSc.
Primary data associated with the manuscript 10.1126/science.adi9926 (“Rab29-dependent asymmetrical activation of leucine-rich repeat kinase 2”)
By Julia Julia Leonard onPrimary data associated with the manuscript "Rab29-dependent asymmetrical activation of leucine-rich repeat kinase 2" (Hanwen Zhu, Francesca Tonelli, Martin Turk, Alan Prescott, Dario R. Alessi, Ji Sun). These include immunoblotting data and their quantitation, representative confocal microscopy images, and quantitation and statistical analysis of confocal microscopy images.
Peripheral MC1R activation modulates immune responses and confers neuroprotection in a mouse model of Parkinson’s disease
By Emma Sherrell onDataset corresponding to the article, "Peripheral MC1R activation modulates immune responses and confers neuroprotection in a mouse model of Parkinson's disease."
Nigrostriatal organotypic cultures to study neuromelanin accumulation in dopaminergic circuits
By Julia Julia Leonard onIn this protocol, the authors describe the preparation and maintenance of rat organotypic cultures from parasagittal brain slices.
FASTQ alignment, gene counts, and differential expression analysis and functional annotation of DEGs
By Julia Julia Leonard onFASTQ alignment, gene counts, and differential expression analysis and functional annotation of DEGs.
Patient PBMC differentiation to macrophages
By Julia Julia Leonard onPatient PBMC differentiation to macrophages.
