pCAG-OSF-ATG13 (2-197)-K15D
By onPlasmid for expression of human ATG13 HORMA K15D mutant in mammalian cells.
Determination of NM Concentration
By onThis is the protocol for determining neuromelanin concentration and data.
cDNA clones for expression of LysoTag (TMEM192) in mammalian cells
By onPlasmids for expression of TMEM192 (LysoTag) in human and mouse cells.
Immunohistochemistry for Carbon Fiber Thread Electrodes
By onMethods for immunofluorescent staining of brain tissue with indwelling electrodes.
A step forward for LRRK2 inhibitors in Parkinson’s disease
By onIn common with the majority of neurodegenerative diseases, there is an urgent and pressing need for novel disease modifying therapies for Parkinson’s disease (PD). Reporting the results of the first human trial for kinase inhibitors of Leucine Rich Repeat Kinase 2 (LRRK2), Jennings and co-workers presented an important advance along the drug development pathway for a target that has long been a priority for the Parkinson’s research community. The focus discusses several topics including: functional characterisation of LRRK2 and the impact of mutations and a key role for altered kinase function in disease; the human genetics of the LRRK2 locus; the outcome of a first-in-human clinical trial for LRRK2 kinase inhibitors by Denali therapeutics, LRRK2 kinase inhibition as a therapeutic strategy in humans; the strategy of using antisense oligonucleotide knockdown approach and the challenges faced by clinical trials – measuring outcomes in chronic, slowly progressing disorders with variable rates of progression.
pCAG- WIPI2dC93E-cs- TEV-STREP
By onPlasmid: Mammalian expression of human WIPI2d C93E with C-terminal Strep.
Immunofluorescent staining for neuronal marker MAP2
By onThis is the protocol for immunofluorescent staining for neuronal marker MAP2.
3D-correlative FIB-milling and Cryo-ET of Autophagic structures in Yeast Cells
By onThis protocol describes how to plunge-freeze yeast on EM grids and how to target autophagic structures by combining cryo confocal fluorescence data to FIB-milling and tomogram acquisition.
Expansion Microscopy
By onExpansion microscopy is a technique to visualize biological structures with higher spatial resolution than traditional microscopy methods.
Microscopy-based GSH bead protein-protein interaction assay
By onProtocol describing the Microscopy-based GSH bead protein-protein interaction assay
Unconventional Initiation of PINK1/Parkin Mitophagy by Optineurin
By onCargo sequestration is a fundamental step of selective autophagy in which cells generate a double membrane structure termed an autophagosome on the surface of cargoes. The authors uncover an unconventional path of PINK1/Parkin mitophagy initiation.
circRNA dataset
By onNew data set on circRNA in laser-captured neuron samples from 190 human brains of healthy controls, prodromal PD, and clinical PD (Dong et al., Nature Communications, in press). The RNA-seq raw FASTQ data and normalized expression matrix of all circRNAs in this study have been deposited in GEO under accession number GSE218203.
circRNA Custom Code
By onCustom code associated with (Dong et al., Nature Communications, in press) is publicly available at https://github.com/sterding/circRNA.
Mutations in Parkinsonism-linked endocytic proteins synaptojanin1 and auxilin have synergistic effects on dopaminergic axonal pathology
By onParkinson's disease (PD) is a neurodegenerative disorder characterized by defective dopaminergic (DAergic) input to the striatum. Mutations in two genes encoding synaptically enriched clathrin-uncoating factors, synaptojanin 1 (SJ1) and auxilin, have been implicated in atypical Parkinsonism. SJ1 knock-in (SJ1-KIRQ) mice carrying a disease-linked mutation display neurological manifestations reminiscent of Parkinsonism. Here we report that auxilin knockout (Aux-KO) mice display dystrophic changes of a subset of nigrostriatal DAergic terminals similar to those of SJ1-KIRQ mice. Furthermore, Aux-KO/SJ1-KIRQ double mutant mice have shorter lifespan and more severe synaptic defects than single mutant mice. These include increase in dystrophic striatal nerve terminals positive for DAergic markers and for the PD risk protein SV2C, as well as adaptive changes in striatal interneurons. The synergistic effect of the two mutations demonstrates a special lability of DAergic neurons to defects in clathrin uncoating, with implications for PD pathogenesis in at least some forms of this condition.
pBPK1520-SNCA-A53T-ng
By onIt can be used to introduce SNCA-A53T mutation using prime editing, PE3 approach.
Live-cell imaging for synaptic vesicle precursors in human iNeuron axons
By onThe authors describe procedure and equipment used for live-imaging of synaptic vesicle precursors.
