Detecting rhythmic spiking through the power spectra of point process model residuals
By savannah onSingle-unit recording is the gold standard for characterizing neurophysiological activity in the basal ganglia and interconnected structures. The work highlighted in this manuscript improves our ability to characterize oscillatory activity and detect pathological changes at the level of individual neurons.
Astrocytic LRRK2 controls synaptic connectivity through ERM phosphorylation
By savannah onThis work reveals a causal link between the astrocytic and synaptic dysfunction induced by the PD-linked mutation, LRRK2 G2019S, providing a non-cell autonomous mechanism for rescuing aberrant cortical wiring
Testing AAVrg-hSyn-mCherry in the spinal cord of Rhesus Macaques
By Emma Sherrell onViral Vector: Two rhesus macaque animals were tested for AAVrg-hSyn-mCherry in the spinal cord. The authors observed abundant mCherry-positive neurons in expected targets (i.e., motor cortical regions known to project to the spinal cord).
Preparation of soluble and insoluble mitochondrial protein fractions for mass spectrometry analysis
By Emma Sherrell onPreparation of soluble and insoluble mitochondrial protein fractions from HeLa cells for mass spectrometry analysis.
Sex distribution of GBA1 variant carriers with dementia with Lewy Bodies and Parkinson’s disease
By savannah onSex distribution of GBA1 variant carriers with dementia with Lewy Bodies and Parkinson's disease.
Primary data associated with the manuscript 10.1073/pnas.2315171120 (“Localization of PPM1H phosphatase tunes Parkinson’s disease-linked LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis”)
By Emma Sherrell onPrimary data associated with the manuscript 10.1073/pnas.2315171120 ("Localization of PPM1H phosphatase tunes Parkinson’s disease-linked LRRK2 kinase-mediated Rab GTPase phosphorylation and ciliogenesis").
Transfection by electroporation of GFP-LRRK2 and Immunofluorescent imaging of MEFs VPS35 (D620N) mutants stably expressing LysoTag
By Emma Sherrell onThe authors investigated the colocalization of GFP-LRRK2 with a lysosomal localized TMEM192-3xHA.
Generation of A549 PPM1H BromoTag CRISPR/CAS9 knock-in cell line
By Emma Sherrell onThis protocol details the generation of A549 PPM1H BromoTag CRISPR/CAS9 knock-in cell line.
Video tutorials for Curtain and Curtain-PTM
By Emma Sherrell onVideo tutorials to assist users of Curtain and Curtain-PTM proteomic data analysis web tools (described in https://www.biorxiv.org/content/10.1101/2023.07.25.550405v1).
Immunoblots
By Emma Sherrell onThe steps of this protocol are similar for different sources of protein such as cell lysate, cytosol, or membrane samples, and EV samples. However, some specific changes depend on the protein of interest, endogenous alpha-synuclein, or type of cells, hiPSC dopamine neurons.
shRNA knockdown
By Emma Sherrell onThis protocol describes a standard procedure to generate a stable cell line for a DNAJC5 knockdown using shRNA.
Differentiation of SH-SY5Y cells
By Emma Sherrell onThis protocol describes a standard procedure to differentiate SH-SY5Y cells into dopaminergic neurons using retinoic acid.
Mitochondria purification
By Emma Sherrell onThis protocol describes how to purify mitochondria from cell culture.
Co-immunoprecipitation
By Emma Sherrell onThis protocol describes a common procedure to perform co-immunoprecipitation.
Medium fractionation and EV preparation
By Emma Sherrell onThis protocol describes the characterization of the extracellular alpha-synuclein and DNAJC5.
CRISPR/Cas9 genome editing
By Emma Sherrell onThis protocol describes the generation of DNAJC5 KO using CRISPR/Cas9 edition.
In vitro depalmitoylation assay
By Emma Sherrell onThis protocol describes the in vitro depalmitoylation assay of DNAJC5.
Membrane and cytosol fractionation
By Emma Sherrell onThis protocol describes membrane and cytosol fractionation of cells expressing different DNAJC5 isoforms.
LRRK2 kinase inhibition reverses G2019S mutation-dependent effects on tau pathology spread
By Emma Sherrell onPreprint: Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson’s disease (PD). This work supports a protective role of LRRK2 kinase inhibition in G2019S carriers and provides a rational workflow for systematic evaluation of brain-wide phenotypes in therapeutic development.
A topographical atlas of αSyn dosage and cell-type expression in the mouse brain and periphery
By Emma Sherrell onPreprint: Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide and presents pathologically with Lewy pathology and dopaminergic neuron loss. This atlas provides much-needed insight into the cellular topography of αSyn, and provides a quantitative map to test assumptions about the role of αSyn in network vulnerability in PD and other αSynucleinopathies.