ATG9A-vesicle isolation protocol
By savannah onThis protocol details the ATG9-vesicle isolation procedure from Hap1 cells.
Immunofluorescence and confocal microscopy
By savannah onThis protocol details the process of immunofluorescence and confocal microscopy.
pLKO.1 mTagBFP2 rat PGK1 shRNA
By savannah onExpresses mTagBFP2 along with an shRNA against rat PGK1.
Protocol for quantification of the nigrostriatal system in non-human primate brain sections
By Emma Sherrell onThe intraputaminal delivery of AAV9-SynA53T resulted in a widespread synucleinopathy throughout the cerebral cortex and substantia nigra in the non-human primate brain.
DFP-induced mitophagy experiments
By savannah onThis protocol details how to induce mitophagy with DFP.
Protocol for ventriculography-assisted stereotaxic surgery in non-human primates
By Emma Sherrell onA step-by-step procedure describing ventriculography-assisted stereotaxic surgery in non-human primates is provided here, including (i) animal pre-surgical preparation, (ii) stereotaxic surgery, and (iii) post-surgical care. This protocol is intended to be used for intraparenchymal deliveries of viral suspensions.
Reconstitution of BNIP3/NIX-mediated autophagy reveals two pathways and hierarchical flexibility of the initiation machinery
By savannah onDuring selective autophagy transmembrane, cargo receptors can trigger autophagy by recruiting different complexes and utilizing multiple pathways. This flexibility in autophagy initiation has important therapeutic implications.
Protocol for non-human primate necropsy
By Emma Sherrell onHere, the authors describe the standard procedure conducted for performing a non-human primate necropsy, followed by removal of the brain and spinal cord from the skull and backbone, respectively.
Cortico-amygdala synaptic structural abnormalities produced by templated aggregation of α-synuclein
By savannah onPD and DLB involve α-syn inclusions in the amygdala affecting cognition and emotions. Cortico-amygdala synapses with α-syn aggregates show increased volume of synapses, potentially contributing to behavioral impairments.
Protocol for immunoperoxidase detection of alpha-synuclein in non-human primate brain samples
By Emma Sherrell onHere, a step-by-step protocol is provided summarizing the immunoperoxidase detection of neurons expressing alpha-synuclein in the macaque brain. The enclosed image illustrates layer V pyramidal neurons showing immunoreactivity for alpha-synuclein at the level of the cerebral cortex (insular gyrus).
α-Synuclein protein preparation (Large scale)
By savannah onProtocol for large-scale preparation of recombinant α-synuclein protein.
Protocol for neuromelanin accumulation drives endogenous synucleinopathy in non-human primates
By Emma Sherrell onCollection of protocols used in this publication.
Culturing 3T3 cells for validating GEARBOCS constructs
By savannah onThis protocol is for culturing Cas9 NIH/3T3 cells for the validation of GEARBOCS 2.0 and similar constructs for use in in vivo assays.
Development and characterization of a non-human primate model of disseminated synucleinopathy
By Emma Sherrell onThe intraputaminal delivery of AAV9-SynA53T resulted in a widespread synucleinopathy throughout the cerebral cortex and substantia nigra in the non-human primate brain.
R scripts associated with SpatialBrain.org
By savannah onAll original R code written to analyze results shown in SpatialBrain.org and Kilfeather & Koo et al. (2024), "Single-cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, aging, and disease."
GEARBOCS: An adeno associated virus tool for in vivo gene editing in astrocytes
By Emma Sherrell onGEARBOCS is a non-invasive method for precise gene editing in astrocytes using CRISPR/Cas9. It allows knockout, tagging, and reporter knock-in strategies, revealing the roles of Sparcl1 and Vamp2 in synapse maintenance in the mouse visual cortex.
Immunohistochemistry for tyrosine hydroxylase (TH), a marker of DA neurons, in mouse brain sections
By savannah onThis protocol describes immunohistochemistry steps for TH staining, a marker of dopaminergic neurons. It was used to confirm the localization of eGFP-tagged ribosomes.
pLKO.1 mTagBFP2 rat DJ-1 shRNA
By savannah onExpresses mTagBFP2 along with an shRNA against rat DJ-1.
