Evaluating endocytic rate in cells
By onAssess endocytic rate in cells using tagged transferrin (Alexa647) and flow cytometry readout.
H9 ES AAVS1-NGN2 TEX264-/-; PiggyBac-Keima-REEP5
By onES cells modified with CRISPR/Cas9 to lack TEX264, express Keima-REEP5 ER-phagy flux reporter, NEUROG2, and mKeima fluorescent protein. Derived from human embryonic stem cells at the blastocyst stage.
H9 ES AAVS1-NGN2 FLAG-EEA1::TMEM192-HA
By onES cells were modified to produce iNeurons expressing FLAG-tagged EEA1 and TMEM192-HA. NEUROG2 construct was inserted into AAVS1 locus using CRISPR/Cas9. EEA1 was N-terminally tagged with a 3xFLAG tag.
Preparing feeder-free hPSCs for nucleofection
By onThis protocol describes the standard procedure preparing feeder-free human pluripotent stem cells (hPSCs) for nucleofection.
Regulatory imbalance between LRRK2 kinase, PPM1H phosphatase, and Arf6 GTPase disrupts the axonal transport of autophagosomes
By onThe authors found that LRRK2-hyperphosphorylated RABs disrupt the axonal transport of autophagosomes by perturbing the coordinated regulation of cytoplasmic dynein and kinesin motors.
CRISPR-Cas9 screen in NIH-3T3 cells to identify modulators of LRRK2 function
By onThis protocol describes a pooled, CRISPR Cas9 screen to identify modulators of LRRK2 activity.
Sample preparation for TMT-based total and phospho-proteomic analysis of cells and tissues
By onMass spectrometry-based proteomics and phosphoproteomics are highly sensitive and unbiased techniques to study the proteome and phosphoproteome at a global scale.
Western blot protocol for detecting ATP10B in mouse/rat brain
By onProtocol for detection of ATP10B in rat and mouse brain tissue by Western blotting.
Unconventional secretion of α-synuclein mediated by palmitoylated DNAJC5 oligomers
By onThe secretion of endogenous α-syn mediated by DNAJC5 is found in a human neuroblastoma cell line, The authors propose that DNAJC5 forms a palmitoylated oligomer to accommodate and export α-syn.
Flow cytometry-based measurement of mitophagic flux
By onAssociated with preprint: https://www.biorxiv.org/content/10.1101/2022.11.02.514817v1
Impaired dopamine release in Parkinson’s disease
By onEvidence indicates that impaired dopamine release can result from disruption to a diverse range of Parkinson’s disease-associated genetic and molecular disturbances and can be considered a potential pathophysiological hallmark of Parkinson’s disease.
SpatialBrain.org – a platform to view integrated data from Wade-Martins Laboratory of Molecular Neurodegeneration
By onAn open repository to access data and results from spatial and translatome expression profiling of basal dopaminergic neurons in mice (OVX and aging). More information can be viewed in published article Kilfeather & Khoo, et al. (2023) "Single-cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, aging, and disease" Spatial Transcriptomic Analyses: - Cell Type Markers - SN/VTA Markers in Dopaminergic Neurons - Cell Number Changes in Age TRAP Analyses: - Dopaminergic Markers - Dopaminergic Ageing - Alternative Splicing in Dopaminergic Neurons Coming soon: Integration with iPS-derived dopaminergic transcriptomic datasets.
Modelling human brain-wide pigmentation in rodents recapitulates age-related multisystem neurodegenerative deficits
By onAuthors describe an animal model of human-like neuromelanin pigmentation. The animals display age-related neuronal dysfunction and degeneration affecting numerous brain circuits and body tissues.
Python script for Golgi cytoscape analysis
By onCustom Python script used for the cytoscape network analysis reported in doi.org/10.1101/2022.11.22.517583 (Golgi-IP, a novel tool for multimodal analysis of Golgi molecular content).
POLCAM: Instant molecular orientation microscopy for the life sciences
By onPOLCAM is a simplified single-molecule orientation localization microscopy method, using a polarization camera, enabling fast easy implementation on fluorescence microscopes. Here, we apply it to alpha-synuclein fibrils.
