AAVS1 Knock-in
By onThis protocol describes the standard procedure to knock-in constructs to the AAVS1 safe harbor locus in hPSCs.
Neuromelanin accumulation drives endogenous synucleinopathy in non-human primates
By onA new NHP PD model was developed by inducing neuromelanin (NM) accumulation in dopaminergic neurons, leading to synucleinopathy and neuron degeneration similar to human PD neuropathology. Lowering NM levels may offer PD therapeutic strategies.
Crystal structure ATG9 HDIR in complex with the ATG13:ATG101 HORMA dimer
By onCrystal structure ATG9 HDIR in complex with the ATG13:ATG101 HORMA dimer (Method: X-RAY DIFFRACTION Resolution: 2.41 Å R-Value Free: 0.251 R-Value Work: 0.204 R-Value Observed: 0.205)
LRRK2 regulates the activation of the unfolded protein response and antigen presentation in macrophages during inflammation
By onAbsence of PINK1 leads to MitAP over-activation engaging autoimmune mechanisms. This pathway is induced by TLR4, cGAS-STING, and UPR activation in response to inflammatory signals. LRRK2 and STING regulate transition from innate to adaptive immunity
Genome-wide determinants of mortality and motor progression in Parkinson’s disease
By onThe authors examined the impact of gene variants on mortality and cognitive impairment in PD. Only the non-Gaucher disease causing GBA PD risk variant E326K, of the known PD risk variants, was associated with progression in PD.
Protocol for hippocampal neuronal cultures
By onThis protocol details the procedue for preparation of neuronal culturs from mice hippocampi as it was performed in Guillén-Samander et al. 2021 article "VPS13D bridges the ER to mitochondria and peroxisomes via Miro" but can also be used to prepare culturs of cortical neurons.
Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism – ASAP protocol collection
By onCollection of protocols of Deleidi Lab used in the publication: "Glucocerebrosidase is imported into mitochondria and preserves complex I integrity and energy metabolism."
Regulation of mitophagy by the NSL complex underlies genetic risk for Parkinson’s disease at 16q11.2 and MAPT H1 loci
By onImpaired mitophagy is linked to familial PD. New study identifies KAT8 and KANSL1 as regulators of PINK1-dependent mitophagy, suggesting their role in idiopathic Parkinson’s. KANSL1 may be crucial in the disease, offering potential drug targets.
Central and Peripheral Inflammation: Connecting the Immune Responses of Parkinson’s Disease
By onAuthors highlight the important work being done that implicates central and peripheral inflammation in playing a role in PD.
Biochemical detection of aggregated Tau
By onThis protocol is associated with the following preprint, published on February 19th 2022: The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds Itika Saha, Patricia Yuste-Checa, Miguel Da Silva Padilha, Qiang Guo, Roman Körner, Hauke Holthusen, Victoria A. Trinkaus, Irina Dudanova, Rubén Fernández-Busnadiego, Wolfgang Baumeister, David W. Sanders, Saurabh Gautam, Marc I. Diamond, F. Ulrich Hartl, Mark S. Hipp bioRxiv 2022.02.18.481043; doi: https://doi.org/10.1101/2022.02.18.481043
Neuropathological Features of Gaucher Disease and Gaucher Disease with Parkinsonism
By onDeficient acid β-glucocerebrosidase activity due to biallelic mutations in GBA1 results in Gaucher disease (GD). Patients with this lysosomal storage disorder exhibit a wide range of associated manifestations, spanning from virtually asymptomatic adults to infants with severe neurodegeneration. While type 1 GD (GD1) is considered non-neuronopathic, a small subset of patients develop parkinsonian features. Variants in GBA1 are also an important risk factor for several common Lewy body disorders (LBDs). Neuropathological examinations of patients with GD, including those who developed LBDs, are rare. GD primarily affects macrophages, and perivascular infiltration of Gaucher macrophages is the most common neuropathologic finding. However, the frequency of these clusters and the affected anatomical region varies. GD affects astrocytes, and, in neuronopathic GD, neurons in cerebral cortical layers 3 and 5, layer 4b of the calcarine cortex, and hippocampal regions CA2–4. In addition, several reports describe selective degeneration of the cerebellar dentate nucleus in chronic neuronopathic GD. GD1 is characterized by astrogliosis without prominent neuronal loss. In GD-LBD, widespread Lewy body pathology is seen, often involving hippocampal regions CA2–4. Additional neuropathological examinations in GD are sorely needed to clarify disease-specific patterns and elucidate causative mechanisms relevant to GD, and potentially to more common neurodegenerative diseases.
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.
Light microscopy immunoperoxidase staining
By onProtocol for Light microscopy immunoperoxidase staining.
Preparation of LRRK1 RCKW cryo-EM grids
By onProtocol used to create LRRK1 RCKW grids for cryo-EM used in Snead, Matyszewski, Dickey et al.
Protocol Collection for “PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons and HeLa cell systems”
By onCollection of protocols associated with preprint (https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1.full) Protocols: Microscopy-based evaluation of Parkin translocation and mitophagy in FBXO7-/- cell lines Microscopy-based pUb-coverage measurements of mitochondria in iNeurons Microscopy-based measurements of p62 recruitment in HeLa Microscopy-based mtDNA turnover measurements in HeLa and iNeurons Microscopy-based mitochondrial morphology measurements in iNeurons
Protocols for “Synaptotagmin-1-dependent phasic axonal dopamine release is dispensable for basic motor behaviors in mice”
By onThis is a collection of protocols for the manuscript "Synaptotagmin-1-dependent phasic axonal dopamine release is dispensable for basic motor behaviors in mice"
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.
