Neuronal hyperactivity-induced oxidant stress promotes in vivo α-synuclein brain spreading
By onInterneuronal transfer and brain spreading of pathogenic proteins are features of neurodegenerative diseases. Pathophysiological conditions and mechanisms affecting this spreading remain poorly understood. This study investigated the relationship between neuronal activity and interneuronal transfer of α-synuclein, a Parkinson-associated protein, and elucidated mechanisms underlying this relationship. In a mouse model of α-synuclein brain spreading, hyperactivity augmented and hypoactivity attenuated protein transfer. Important features of neuronal hyperactivity reported here were an exacerbation of oxidative and nitrative reactions, pronounced accumulation of nitrated α-synuclein, and increased protein aggregation. Data also pointed to mitochondria as key targets and likely sources of reactive oxygen and nitrogen species within hyperactive neurons. Rescue experiments designed to counteract the increased burden of reactive oxygen species reversed hyperactivity-induced α-synuclein nitration, aggregation, and interneuronal transfer, providing first evidence of a causal link between these pathological effects of neuronal stimulation and indicating a mechanistic role of oxidant stress in hyperactivity-induced α-synuclein spreading.
Molecular cloning of SHIP164 plasmids for expression in mammalian cells
By onThis protocol is to help with the molecular cloning of SHIP164 and the sequences of other proteins.
H9 ES AAVS1-NGN2 CCPG1-/-; PiggyBac-Keima-REEP5
By onES cells were modified using CRISPR/Cas9 to lack CCPG1 and express Keima-REEP5 ER-phagy flux reporter. NEUROG2 construct was introduced in AAVS1 locus. Cells are embryonic stem cells derived from a human female blastocyst stage.
Preparing MEF-cultured hPSCs for nucleofection
By onThis protocol describes the procedure or preparing MEF-cultured human pluripotent stem cells (hPSCs) for the delivery of plasmids, mRNA, or ribonucleoprotein (RNP) using nucleofection.
The remote assessment of parkinsonism supporting the ongoing development of interventions in Gaucher disease
By onMutations in GBA which are causative of Gaucher disease in their biallelic form, are the most common genetic risk factor for Parkinson's disease (PD). The diagnosis of PD relies upon clinically defined motor features which appear after irreversible neurodegeneration. Prodromal symptoms of PD may provide a means to predict latent pathology, years before the onset of motor features. Previous work has reported prodromal features of PD in GBA mutation carriers, however this has been insufficiently sensitive to identify those that will develop PD. The Remote Assessment of Parkinsonism Supporting Ongoing Development of Interventions in Gaucher Disease (RAPSODI GD) study assesses a large cohort of GBA mutation carriers, to aid development of procedures for earlier diagnosis of PD.
LRRK2-G2019S Synergizes with Ageing and Low-Grade Inflammation to Promote Gut and Peripheral Immune Cell Activation that Precede Nigrostriatal Degeneration
By onBackground Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most frequent cause of familial Parkinson’s disease (PD). The incomplete penetrance of LRRK2 mutations suggest that additional hits are required for disease onset. We hypothesized that chronic low-grade inflammation interacts with LRRK2 G2019S, the most frequent PD-associated mutation, to activate peripheral and central immune reactions and drive age-dependent neurodegeneration. Methods and Results We exposed wild-type and LRRK2 G2019S mice to a low chronic dose of lipopolysaccharide, and we performed a longitudinal analysis of central and peripheral immune reactions and neurodegeneration. Low-dose inflammation triggered nigrostriatal degeneration, macrophage/monocyte brain infiltration, and astro-/microgliosis. LRRK2 G2019S mice showed an early dysregulation of peripheral cytokines, increased CD4+ T-cell infiltration and α-synuclein aggregation in the colon. Interestingly, peripheral immune activation and colonic α-synuclein aggregation precede astro-/microgliosis and neurodegeneration. Conclusions Our study suggests an early role of the peripheral immune system and the gut in LRRK2 PD and provides a novel model to study early therapeutic immune targets and biomarkers.
Reconstitution of cargo-induced LC3 lipidation in mammalian selective autophagy
By onSelective autophagy is essential for maintaining cellular homeostasis. Using in vitro reconstitution, the authors explored the details of mitophagy initiation from autophagy receptor engagement through LC3 lipidation.
Microscopy-based bead protein-protein interaction assay
By onAn assay to study protein-protein interaction
Dopaminergic axons track somatic signaling in behaving mice
By onHere, authors used genetic strategies to isolate key dopaminergic neuron subtypes and monitor their axonal and somatic signaling patterns in behaving mice.
Live imaging to investigate mitophagy kinetics and NEMO recruitment in HeLa-M cells
By onProtocol for live imaging to investigate mitophagy kinetics and NEMO recruitment in HeLa-M cells.
Gene editing of YIPF4 in hESCs V3
By onThis protocol describes the creation of YIPF4 knockout cell lines in H9 hESC cells using CRISPR-Cas9. Associated with preprint: https://doi.org/10.1101/2022.12.06.519342
Lentivirus production
By onThis protocol is associated with the following preprint, published on February 19, 2022: “The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds.”
Striatal dopamine measurement through HPLC
By onProtocol for striatal dopamine measurement in mouse brain
Primary data associated with the manuscript “Whole proteome copy number dataset in primary mouse cortical neurons” (doi: 10.1016/j.dib.2023.109336)
By onRaw data reported in Supplementary table in "Whole Proteome Copy Number Dataset in Primary Mouse Cortical Neurons."
