pCAG-MBP-Foldon-ATG9 (828-839)
By onPlasmid: To make ATG9 C-terminal tail trimer for expression in mammalian cells.
Primary neuron culture for live imaging of axonal cargoes
By onThis protocol describes the preparation and culture of mouse primary cortical neurons for live-imaging experiments. Neurons were transfected 16-24 hours before imaging using Lipofectamine 2000.
Neuromelanin staining (Fontana-Masson staining)+ TH-DAB staining on midbrain organoids
By onNeuromelanin staining (Fontana-Masson staining)+ TH-DAB staining on midbrain organoids
Primary neuronal cultures
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
Local genetic correlations exist among neurodegenerative and neuropsychiatric diseases
By onGenetic correlation between neurodegenerative and neuropsychiatric diseases was explored using local rg analysis. Unique relationships were found, suggesting shared genetic mechanisms and potential therapeutic targets in complex diseases.
Who is at Risk of Parkinson Disease? Refining the Preclinical Phase of GBA1 and LRRK2 Variant Carriers: a Clinical, Biochemical, and Imaging Approach
By onPurpose of Review Genetic variants in GBA1 and LRRK2 genes are the commonest genetic risk factor for Parkinson disease (PD); however, the preclinical profile of GBA1 and LRRK2 variant carriers who will develop PD is unclear. This review aims to highlight the more sensitive markers that can stratify PD risk in non-manifesting GBA1 and LRRK2 variant carriers. Recent Findings Several case–control and a few longitudinal studies evaluated clinical, biochemical, and neuroimaging markers within cohorts of non-manifesting carriers of GBA1 and LRRK2 variants. Summary Despite similar levels of penetrance of PD in GBA1 and LRRK2 variant carriers (10–30%), these individuals have distinct preclinical profiles. GBA1 variant carriers at higher risk of PD can present with prodromal symptoms suggestive of PD (hyposmia), display increased α-synuclein levels in peripheral blood mononuclear cells, and show dopamine transporter abnormalities. LRRK2 variant carriers at higher risk of PD might show subtle motor abnormalities, but no prodromal symptoms, higher exposure to some environmental factors (non-steroid anti-inflammatory drugs), and peripheral inflammatory profile. This information will help clinicians tailor appropriate screening tests and counseling and facilitate researchers in the development of predictive markers, disease-modifying treatments, and selection of healthy individuals who might benefit from preventive interventions.
pCAG-MBP-ATG9 (692-839)
By onPlasmid: To make monomeric ATG9 C-terminal tail for expression in mammalian cells.
The mass spectrometry proteomics data associated of TauRD-Y interactome from TauRD-Y and TauRD-Y* cells.
By onThe mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD023400. Associated with the following preprint:
Microscopy-based evaluation of mtKeima flux in hESC-derived Ctrl and FBXO7-/- iNeurons
By onProtocol for the microscopy-based evaluation of mtKeima flux in hESC-derived Ctrl and FBXO7-/- iNeurons PARK15/FBXO7 is dispensable for PINK1/Parkin mitophagy in iNeurons and HeLa cell systems: https://www.embopress.org/doi/full/10.15252/embr.202256399
Thawing of hPSCs grown on MEFs
By onThis protocol describes the standard procedure of thawing human pluripotent stem cells (hPSCs) on inactivated mouse embryonic fibroblasts (MEFs).
pGBdest-10xHis-TEV-ATG5_ATG7_ATG10_ATG12_ATG16L1_F32A-I35A-I36A_-GFP-TEV-StrepII
By onPlasmid: Plasmid for the Expression of the E3 complex with an FII (F32A, I35A, and I36A) mutation. SMC Internal reference: SMC1728
Hydrop enables droplet-based single-cell ATAC-seq and single-cell RNA-seq using dissolvable hydrogel beads
By onThe data available in this repository can be used to replicate all the figures in the authors’ manuscript using their data analysis tutorial available at https://github.com/aertslab/hydrop_data_analysis.
Stereology-mediated cell counting using StereoInvestigator
By onProtocol for stereologic count with StereoInvestigator.
Genotyping by next generation sequencing
By onThis collection describes a standard genotyping procedure using next generation sequencing (NGS) in the Hockemeyer lab.
Proteostasis and lysosomal quality control deficits in Alzheimer’s disease neurons
By onLysosomal quality control (LQC) pathways are notably impaired in both aging and AD, leading to neuronal vulnerability and cytotoxicity. Neurons show amyloid-β inclusions, and enhancing lysosomal function can help alleviate AD-related pathologies.
pLenti HsATP10B_D433N-T2A-His-flag-TMEM30A
By onPlasmid: Transfer plasmid for lentiviral vector production expressing Hs ATP10B D433N mutant and His/flag tagged Hs TMEM30A.
pCAG- WIPI2dK88E-cs- TEV-STREP
By onPlasmid: Mammalian expression of human WIPI2d K88E with C-terminal Strep.
ALS and FTD-associated missense mutations in TBK1 differentially disrupt mitophagy
By onSource data for manuscript https://doi.org/10.5281/zenodo.4670341
H9 ES AAVS1-NGN2 FAM134C/A/B-/-;TEX264-/-; PiggyBac-Keima-RAMP4
By onES cells were modified to create iNeurons lacking ER-phagy receptors FAM134C, A, B, TEX264, and expressing Keima-RAMP4 ER-phagy flux reporter. CRISPR/Cas9 was used to introduce NEUROG2 construct in AAVS1 locus and knockout RETREG and TEX264 genes.
