Basal Ganglia Neurons in Healthy and Parkinsonian Primates Generate Recurring Sequences of Spikes
By onData consists of interspike intervals from basal ganglia neurons in awake monkeys, pre- and post-parkinsonian treatment with MPTP. Recordings were during spontaneous spiking, without task involvement.
Spatial transcriptomics reveals molecular dysfunction associated with Lewy pathology
By onThe results identify neurons vulnerable to Lewy pathology in the PD cortex and identify a conserved signature of molecular dysfunction in both mice and humans.
pCAG-MBP-ATG9-D830A/E831A
By onPlasmid for expression of human ATG9-D830A/E831A mutant in mammalian cells
AAV viral DNA and whole RNA recovery for AAV pool experiments in rhesus macaque
By onThis protocol outlines procedures to extract viral DNA and whole RNA from rhesus macaque tissue that had been treated with AAV in vivo
Lentivirus plasmid: pLV[Exp]-U6>KAT8_P1_Seq1-hPGK>mApple
By onPlasmid: Plasmid vector encoding a sgRNA sequence which targets the human KAT8 promoter for CRISPRi, under a U6 promoter and a mApple fluorescent reporter. Generated by Vectorbuilder in the pLV backbone.
x4 GBA Plasmids
By onThe below plasmids are deposited and available via Addgene:https://www.addgene.org/Anthony_Schapira/ . These have been used in publication: 10.1093/hmg/ddac233 188580 WT GBA pcDNA3.1 GBA (Homo sapiens) 188581 E326K GBA pcDNA3.1 GBA (Homo sapiens) 188582 L444P GBA pcDNA3.1 GBA (Homo sapiens) 188583 N370S GBA pcDNA3.1 GBA (Homo sapiens)
Catalepsy test (Bar test)
By onThe catalepsy test (bar test) was developed to test motor coordination and motor impairments.
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.
Expression and purification of mCherry-OPTN
By onThis protocol describes the purification of mCherry-OPTN.
pCAG- WIPI2dR125E- cs-TEV -STREP
By onPlasmid: Mammalian expression of human WIPI2d R125E with C-terminal Strep
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).
Structural basis for membrane recruitment of ATG16L1 by WIPI2 in Autophagy
By onThe authors showed through structural determination how ATG16L1 and WIPI2 interact and compared the other WIPI proteins showing the variety of mechanisms of membrane recruitment by WIPI proteins.
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.
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.
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
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.
