Visualizing chaperone-mediated multistep assembly of the human 20S proteasome
By Emma Sherrell onDedicated assembly factors orchestrate the stepwise production of molecular machines, including the 28-subunit proteasome core particle (CP) that mediates protein degradation. This article presents cryo-EM reconstructions of seven recombinant human subcomplexes that visualize all five chaperones and the three active site propeptides across a wide swath of the assembly pathway.
Tau aggregation monitored by thioflavin-T (ThT) fluorescence in a plate reader
By Emma Sherrell onThis protocol details how to efficiently monitor Tau aggregation by thioflavin-T fluorescence in a plate reader.
Solid phase binding assay – Clusterin binding to very low-density lipoprotein receptor (VLDLR)
By Emma Sherrell onThis protocol details how to monitor Clusterin binding to the very low-density lipoprotein receptor (VLDLR) by enzyme-linked immunosorbent assays (ELISA) adapted from Leeb et al., 2014.
Amyloid beta (Aβ) aggregates N-terminal labeling
By Emma Sherrell onThis protocol details how to efficiently label protein aggregates at the N-terminal using Amyloid beta as example.
Harvesting and culturing of peritoneal macrophages and ex vivo stimulation
By Julia Julia Leonard onHarvesting and culturing of peritoneal macrophages and ex vivo stimulation with IFNy.
α-Synuclein aggregation monitored by thioflavin-T (ThT) fluorescence in a plate reader
By Emma Sherrell onThis protocol details how to efficiently monitor α-Synuclein aggregation by thioflavin-T fluorescence in a plate reader.
GFP-Clu-tails purification from Escherichia coli cells
By Emma Sherrell onThis protocol details how to purify the fusion protein GFP-Clu-tails from Escherichia coli.
MBP-Clu-tail purification from Escherichia coli cells
By Emma Sherrell onThis protocol details how to efficiently purify the fusion protein Maltose binding protein (MBP)-Clu-tail (204-238) from Escherichia coli.
Rat organotypic cultures for AAV-mediated vital labeling of the extracellular matrix V.2
By Julia Julia Leonard onIn this protocol, the authors describe the preparation and maintenance of rat organotypic cultures (cortico-striatal) for AAV infection and imaging of the extracellular matrix.
Polyglucosan body density in the aged mouse hippocampus is controlled by a novel modifier locus on chromosome 1
By Emma Sherrell onAging can be associated with the accumulation of polyglucosan bodies (PGBs). While PGBs have a detrimental effect on cognition, the underlying mechanism and clinical relevance of age-related PGB accumulation remain unknown. Here, the authors investigated the genetic basis and functional impact of age-related PGB accumulation in mice.
Transcriptional programs mediating neuronal toxicity and altered glial-neuronal signaling in a Drosophila knock-in tauopathy model
By Emma Sherrell onMutations in the gene encoding tau cause autosomal dominant forms of frontotemporal dementia. Prior models of frontotemporal dementia replicate features of the disease but do not recreate the genetic context. This article presents a genetic model of tauopathy, which recapitulates the genetic context and phenotypic features of the disease.
A human development-based protocol for the differentiation of human ESCs into midbrain dopaminergic neurons
By Julia Julia Leonard onProtocol for the differentiation of human embryonic stem cells into midbrain dopaminergic neurons. This protocol recapitulates the activation of developmental pathways known to be important for mDA neuron development and mimics key sequences of events taking place in human midbrain development.
Neuronal dysfunction and gene modulation by non-coding RNA in Parkinson’s disease and synucleinopathies
By Julia Julia Leonard onEmerging evidence suggests that non-coding RNAs (ncRNAs) contribute to molecular events underlying progressive neuronal degeneration. This review discusses recent literature focused on the role of RNA-based mechanisms involved in different aspects of neuronal pathology in Parkinson’s disease and synucleinopathy models.
Purification of recombinant low-density lipoprotein receptor-related protein associated protein 1 (LRPAP1, RAP) from Escherichia coli
By Emma Sherrell onThis protocol details how to efficiently purify low-density lipoprotein (LDL) receptor-related protein associated protein 1 (LRPAP1 or RAP) from Escherichia coli.
Generation of the iPSC line FINi002-A from a male Parkinson’s disease patient carrying compound heterozygous mutations in the PRKN gene
By Julia Julia Leonard onThis article describes the generation of the iPSC line FINi002-A from a male Parkinson's disease patient carrying compound heterozygous mutations in the PRKN gene.
Alpha-synuclein overexpression can drive microbiome dysbiosis in mice
By Emma Sherrell onGrowing evidence indicates that persons living with Parkinson's disease (PD) have a unique composition of indigenous gut microbes. Here, the authors used a transgenic mouse strain, which overexpress wild-type human α-syn to test how the gut microbiome composition responds in this model of PD pathology during aging.
STC-1 passaging and PD associated microbe stimulation protocol
By Emma Sherrell onSTC-1 passaging and PD-associated microbe stimulation protocol.
SOP for RT (Reverse Transcription) Promega kit
By Emma Sherrell onSOP for RT (Reverse Transcription) Promega kit.
Richert/Heterogeneous_SNNs: v1.0.0: Scripts to replicate the results of Gast et al. PNAS 2024
By Julia Julia Leonard onThis release includes configuration files and scripts to replicate the main results presented in Gast et al. PNAS 2024, including Python scripts for the simulation, bifurcation analysis, and training of networks of Izhikevich neurons with heterogeneous spike thresholds.
SOP for immune cells isolation mouse adult brain with CD45 beads
By Emma Sherrell onSOP for immune cells isolation mouse adult brain with CD45 beads.