APEX-autophagy cargo receptor proteomics with and without lysosomal damage
By onProteomics data associated with DOI: 10.7554/eLife.72328
RAB3 phosphorylation by pathogenic LRRK2 impairs trafficking of synaptic vesicle precursors
By onThe authors’ results reveal a mechanism by which pathogenic hyperactive LRRK2 may contribute to the altered synaptic homeostasis associated with characteristic non-motor and cognitive manifestations of PD.
Catalepsy test (Bar test)
By onThe catalepsy test (bar test) was developed to test motor coordination and motor impairments.
Brain Repair by Cell Replacement via In Situ Neuronal Reprogramming
By onNeurodegenerative diseases, characterized by progressive neural loss, have been some of the most challenging medical problems in aging societies. Treatment strategies such as symptom management have little impact on disease progression, while intervention with specific disease mechanisms may only slow down disease progression. One therapeutic strategy that has the potential to reverse the disease phenotype is to replenish neurons and rebuild the pathway lost to degeneration. Although it is generally believed that the central nervous system has lost the capability to regenerate, increasing evidence indicates that the brain is more plastic than previously thought, containing perhaps the biggest repertoire of cells with latent neurogenic programs in the body. This review focuses on key advances in generating new neurons through in situ neuronal reprogramming, which is tied to fundamental questions regarding adult neurogenesis, cell source, and mechanisms for neuronal reprogramming, as well as the ability of new neurons to integrate into the existing circuitry.
Association between the LRP1B and APOE loci and the development of Parkinson’s disease dementia
By onGenetic analysis of 3,964 PD cases revealed APOE-ϵ4 allele and new loci as risk factors for PD dementia progression, implicating amyloid pathway in PDD development and potential for amyloid-targeting therapy.
Analysis of glycosphingolipids from human plasma
By onThis is an updated protocol with the focus on achieving sensitive and reproducible quantitation of glycosphingolipids from human plasma samples
Plasmid: pLV[Exp]-EF1A>V5/hKAT8[NM_032188.3]:IRES:Puro
By onPlasmid: Human KAT8 gene with N-terminal V5 tag cloned into the pLV plasmid backbone under a EF1a promoter and including IRES puromycin selection cassette by Vectorbuilder.
Cell line construction and maintenance for Lyso-IP and Endo-IP analysis of amyloid precursor protein processing
By onLyso-IP is a method that allows for the isolation of lysosomes for proteomics and metabolomics (dx.doi.org/10.17504/protocols.io.bybjpskn; dx.doi.org/10.17504/protocols.io.bx9hpr36). We have developed an analogous approach for purification of early/sorting endosomes (Endo-IP). In addition, we have found that endolysosomal purification via Lyso-IP and Endo-IP can be coupled with a quantitative proteomics workflow to obtain snapshots of Amyloid Precursor Protein (APP) processing to its Aβ products (Park et al. in submission). Here, we describe methods for cell line construction and maintenance of 293 cells with TMEM192-3xHA and 3xFLAG-EEA1, which are used for lysosome and endosome purification, respectively, with the addition of patient mutations to APP promotes processing. Cells with endogenously tagged TMEM192 and stably expressing FLAG-EEA1 are referred to as 293EL cells, for Endo-IP and Lyso-IP. These cells were also prepared in a form that has a deletion of the APP gene (293EL;APP-/-) and the same cells reconstituted with a lentivirus stably expressing APPSw;T700N to allow functional analysis of APP processing.
pCAG-OSF-ATG13 (2-197)-W50D
By onPlasmid for expression of human ATG13 HORMA W50D mutant in mammalian cells.
Adhesive Removal Test to assess sensorimotor deficits in parkinsonian mice
By onThis behavior is used to assess fine motor movements in a mouse Parkinsonian model. It checks for correct paw and mouth sensitivity (time-to-contact) and correct dexterity (time-to-remove).
Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex
By onAuthors determine cryo-EM structure of human ULK1C core and its complex with PI3KC3-C1. ULK1C core undergoes a rearrangement from 2:1:1 to 2:2:2 stoichiometry, suggesting a structural mechanism for autophagy initiation.
GFP-TBK1: expression and purification
By onThis protocol describes how to express and purify human TBK1 tagged N-terminally with eGFP.
The Hsc70 disaggregation machinery removes monomer units directly from α-synuclein fibril ends
By onUsing microfluidic diffusional sizing, the authors show that the molecular chaperone family Hsp70 (specifically Hsc70, DnaJB, and Apg2) can completely dissolve alpha-synuclein aggregation and revert it back to its monomeric state.
Post mortem human substantial nigra TH staining
By onThis protocol is standardized for postmortem (frozen) SN tissue from pathologically diagnosed PD patients and control individuals for Immunofluorescence staining.
A549 PPM1H-BromoTag (CVCL_C8YV)
By onCell Line: PPM1H-BromoTag CRISPR/CAS9 A549 knock-in cell line generated from parental A549 cells (ATCC A549 CCl-185) by introducing a BromoTag linked to the C-terminus of PPM1H using CRISPR/Cas9 gene-editing technology.
