Assessing autophagy using the HaloTag-LC3B cleavage assay
By onAssay to detect HaloTag-LC3B during starvation autophagy based on https://doi.org/10.7554/eLife.78923.
Generation of stable cell lines using retroviral system
By onThis protocol details the procedure for the generation of stable cell lines using retroviral system.
Raw band intensity values for ATG3 WT or Mutants in vitro LC3 lipidation
By onRaw band intensity values used for the quantification of in vitro LC3 lipidation results (Fig4C) in 10.1101/2023.07.17.549391
Three-step docking by WIPI2, ATG16L1 and ATG3 delivers LC3 to the phagophore: Molecular dynamics simulation dat
By onAtomistic molecular dynamics simulation data set accompanying manuscript 10.1101/2023.07.17.549391.
RASP: Optimal single fluorescent puncta detection in complex cellular backgrounds
By onRASP, a bioimaging-segmentation method, outperforms existing methods by removing false positives + detecting features across various spatial scales. RASP enables precise analysis of cellular and tissue environments, down to single protein levels.
Raw band intensity values for Halo-LC3B
By onSource values referred to raw band intensity used for the quantification of percentage autophagy levels.
VPS13B is localized at the cis-trans Golgi complex interface and is a functional partner of FAM177A1
By onMutations in VPS13B, a member of a protein family implicated in bulk lipid transport between adjacent membranes, cause Cohen syndrome. VPS13B is known to be concentrated in the Golgi complex, but its precise location within this organelle and thus the site(s) where it achieves lipid transport remains unclear. Here we show that VPS13B is localized at the interface between cis and trans Golgi sub-compartments and that Golgi complex re-formation after Brefeldin A (BFA) induced disruption is delayed in VPS13B KO cells. This delay is phenocopied by loss of FAM177A1, a Golgi complex protein of unknown function reported to be a VPS13B interactor and whose mutations also result in a developmental disorder. In zebrafish, the vps13b orthologue, not previously annotated in this organism, genetically interacts with fam177a1. Collectively, these findings raise the possibility that bulk lipid transport by VPS13B may play a role in expanding Golgi membranes and that VPS13B may be assisted in this function by FAM177A1.
napari-polcam
By onA plugin for napari is available to handle and display polarization camera data efficiently.
Lysosomal TBK1 responds to amino acid availability to relieve Rab7-dependent mTORC1 inhibition
By onThis study reveals that TBK1 is recruited to lysosomes when amino acids are abundant where it phosphorylates Rab7 and thus relieves Rab7-dependent suppression of mTORC1 signaling from lysosomes.
A STING-CASM-GABARAP Pathway Activates LRRK2 at Lysosomes
By onMutations that increase LRRK2 kinase activity have been linked to Parkinson’s disease and Crohn’s disease. LRRK2 is also activated by lysosome damage evoked by chemical and pathogenic stimuli. However, the endogenous cellular mechanisms that control LRRK2 kinase activity are not well understood. In this study, we identify signaling through Stimulator of Interferon Genes (STING) as an upstream activator of LRRK2. This LRRK2 activation occurs via the Conjugation of ATG8 to Single Membranes (CASM) pathway. We furthermore establish that multiple chemical stimuli that perturb lysosomal homeostasis also converge on CASM to activate LRRK2. Although CASM mediates the lipidation of multiple ATG8 protein family members, LRRK2 lysosome recruitment and kinase activation is highly dependent on an interaction with the GABARAP member of this family. Collectively these results define a pathway that integrates multiple stimuli at lysosomes to control the kinase activity of LRRK2. Aberrant activation this pathway may be of relevance in both Parkinson’s and Crohn’s diseases.
Mutations in GPNMB associated with Amyloid cutis dyschromica alter intracellular trafficking and processing of GPNMB
By onThis work highlights previously undescribed cellular characteristics of GPNMB missense mutations implicated in ACD and helps to better inform the clinically observed phenotypes, as well as underscore GPNMB’s role at melanosomes.
ICC confocal images: Protein aggregation and calcium dysregulation are the earliest hallmarks of familial Parkinson’s disease in human midbrain dopaminergic neurons
By onUsing hiPSCs, researchers identified early pathophysiological events, including oligomeric aggregate formation, impaired calcium signaling, and mitochondrial dysfunction, ultimately resulting in abnormal neuronal activity and cell death in PD.
Development and characterization of a non-human primate model of disseminated synucleinopathy
By onThe performance and biodistribution of the retrogradely-spreading AAV9-SynA53T vector was evaluated in the NHP brain. Deliveries of viral suspensions in the left putamen gave rise to a disseminated synucleinopathy in a circuit-specific basis.
Neuromelanin accumulation drives endogenous synucleinopathy in non-human primates
By onEvidence shows that intracellular aggregation of endogenous alpha-synuclein is triggered by NMel accumulation. Any therapeutic approach to decrease NMel levels may provide appealing choices for the successful implementation of novel PD therapeutics.
Synchronous Measurements of Extracellular Action Potentials and Neurochemical Activity with Carbon Fiber Electrodes in Nonhuman Primates
By onThe authors developed methods for synchronous measures of neuron spikes and dopamine signals in the monkey striatum. These methods will help advance our understanding of the interactions between neuromodulator signaling and neuronal activity.
Quantification of p62-positive inclusions
By onQuantification of p62-positive inclusions in the rodent brain
