Insect Cell Protocol for LRRK1 and LRRK2 Expression
By onProtocol for expressing LRRK1 and LRRK2 in insect cells.
Cell culture, transfection and imaging
By onThis protocol details the general preparation of cells for imaging and also for imaging experiments involving cellular hypotonic shock and cytosolic Ca2+ changes as they were performed in https://doi.org/10.1083/jcb.202010004.
Halo-LC3B processing assay to assess autophagy
By onThis protocol details Halo-LC3B processing assay to assess autophagy.
A unifying model for the role of the ATG8 system in autophagy
By onThe core ATG8 system is one of the most-studied autophagy components. Here, authors reconcile prior observations of the core ATG8 system into a unifying model.
Tet-off FLTau (HEK293T)
By onAssociated with the following preprint: Saha et al., 2022 (published on biorxiv on Feb 19th, 2022) https://www.biorxiv.org/content/10.1101/2022.02.18.481043v1.full
Astrocyte extraction from brain organoids V.2
By onProtocol for astrocyte extraction from brain organoids.
Therapeutic deep brain stimulation disrupts movement-related subthalamic nucleus activity in Parkinsonian mice
By onSubthalamic nucleus deep brain stimulation (STN DBS) relieves many motor symptoms of Parkinson's Disease (PD), but its underlying therapeutic mechanisms remain unclear. Since its advent, three major theories have been proposed: (1) DBS inhibits the STN and basal ganglia output; (2) DBS antidromically activates motor cortex; and (3) DBS disrupts firing dynamics within the STN. Previously, stimulation-related electrical artifacts limited mechanistic investigations using electrophysiology. We used electrical artifact-free GCaMP fiber photometry to investigate activity in basal ganglia nuclei during STN DBS in parkinsonian mice. To test whether the observed changes in activity were sufficient to relieve motor symptoms, we then combined electrophysiological recording with targeted optical DBS protocols. Our findings suggest that STN DBS exerts its therapeutic effect through the disruption of movement-related STN activity, rather than inhibition or antidromic activation. These results provide insight into optimizing PD treatments and establish an approach for investigating DBS in other neuropsychiatric conditions.
Standard operating procedure for the isolation of genetically engineered hPSCs clones in a high-throughput way
By onThis collection describes a standard procedure for isolating single human pluripotent stem cell (hPSC) clones in a high-throughput way.
Model building, validation, and visualization
By onProtocol describing Model building, validation, and visualization
Proteomics analysis of autophagy cargos reveals distinct adaptations in PINK1 and LRRK2 models of Parkinson disease
By onAutophagy is crucial for neuronal health. Studies on PD mouse models show adaptive pathways are activated to maintain brain balance, despite reduced autophagic flux in PINK1-/- mice and impaired autophagosomal function in LRRK2G2019S mice.
Wire Hang Test
By onThis behavioral test is designed to assess early motor deficits associated with muscle and grip strength.
Bone Marrow Derived Macrophage (BMDM) differentiation and maintenance
By onThis protocol describes the isolation of bone marrow derived macrophages.
Structural basis for ATG9A recruitment to the ULK1 complex in mitophagy initiation
By onHere, the authors examine the structural interaction between ATG9A and components of the ULK1 complex to better understand the process of the PINK1- and Parkin- dependent mitophagy pathway implicated in Parkinson's disease.
Immunocytochemical analysis
By onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Reconstitution of LRRK2 membrane recruitment onto planar lipid bilayers
By onA method to monitor the recruitment of purified LRRK2 kinase onto planar lipid bilayers containing lipid-anchored Rab10 protein using Total Internal Reflection Fluorescence (TIRF) Microscopy.
Large-scale biophysically detailed model of somatosensory thalamocortical circuits in NetPyNE
By onThe work provides a widely accessible, data-driven and biophysically-detailed model of the somatosensory thalamocortical circuits that can be utilized as a community tool for researchers to study neural dynamics, function and disease.
Primary data associated with doi: 10.1042/BCJ20220308 (PKC isoforms activate LRRK1 kinase by phosphorylating conserved residues (Ser1064, Ser1074 and Thr1075) within the CORB GTPase domain)
By onRaw immunoblotting data; immunofluorescence microscopy images; tabular data for quantification; autoradiograph films; scans of Coomassie-stained protein gels; Prism files with statistical analysis.
