Immunofluorescence staining for postmortem mouse brain tissue
By savannah onThis is a basic protocol for staining mouse brain tissues using immunofluorescence and immunohistochemistry techniques.
Genotyping mice from ear clips
By savannah onThis protocol describes the genotyping procedure from ear clip samples. This includes a general PCR protocol for primers with annealing temperatures in the range 55-70 degrees C. For other primers, the thermal cycling should be adjusted.
Endosome isolation
By savannah onSubcellular fractionation to isolate early and late endosomes (EEs and LEs) by performing a series of centrifugation steps.
Create mask for axonal quantification analysis with FIJI
By savannah onThis protocol describes how to create a quantified mask from an image of fluorescence-tagged axonal projections using FIJI/ImageJ software.
Catalepsy test (Bar test)
By savannah onThe catalepsy test (bar test) was developed to test motor coordination and motor impairments.
Adhesive Removal Test to assess sensorimotor deficits in parkinsonian mice
By savannah 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).
Acute striatal or midbrain fiber photometry in head-fixed mice
By savannah onWhile this protocol focuses recordings from striatum with GCaMP, it can be easily modified to record from other brain regions and with other fluorescent reporters.
Vibrational stabilization of complex network systems
By savannah onPublished: Many natural and man-made network systems need to maintain certain patterns, such as working at equilibria or limit cycles, to function properly. The authors provide some numerical results that demonstrate the validity of our theoretical findings.
Parkinson’s genes orchestrate pyroptosis through selective trafficking of mtDNA to leaky lysosomes
By savannah onPreprint: Breach of lysosomes allows mtDNA to access cytosol, requiring multiple Parkinson's disease-related proteins and Gasdermin pores, identified in the screen. These data place mitochondria-to-lysosome transport as a driver of pyroptosis and link multiple PD proteins along a common pathway.
Genetic meta-analysis of levodopa induced dyskinesia in Parkinson’s disease
By savannah onPublished: Based on a functional annotation analysis on chromosome 1, we determined that changes in DNAJB4 gene expression, close to LRP8, are an additional potential cause of increased susceptibility to LiD. Baseline anxiety status was significantly associated with LiD.
Mitochondrial degradation: Mitophagy and beyond
By savannah onPublished: The molecular signals driving varied pathways are discussed, including the cellular and physiological contexts under which the different degradation pathways are engaged.
A RAB7A phosphoswitch coordinates Rubicon Homology protein regulation of Parkin-dependent mitophagy
By savannah onPublished: Structural and functional data support a model in which the TBK1-dependent phosphorylation of RAB7A serves as a switch, promoting mitophagy by relieving Rubicon inhibition and favoring Pacer activation. View original preprint.
Therapeutic potential of PTB inhibition through converting glial cells to neurons in the brain
By savannah onPublished: Multiple lines of evidence suggest that Ptbp1 depletion can convert a selective subpopulation of glial cells into neurons and, via this and other mechanisms, reverse deficits in a Parkinson's disease model, emphasizing the importance of future efforts in exploring this therapeutic strategy.
Endosomal escape of RNA therapeutics: How do we solve this rate-limiting problem?
By savannah onPublished: Over 15 FDA approved drugs, numerous ongoing clinical trials, RNA therapeutics, such as small interfering RNAs (siRNAs) and antisense oligonucleotides (ASOs), have shown potential to treat human disease. RNA therapeutics can be used to treat widespread human disease, the rate-limiting delivery problem of endosomal escape must be solved in a nontoxic manner.
Feed-forward metabotropic signaling by Cav1 Ca2+ channels supports pacemaking in pedunculopontine cholinergic neurons
By savannah onPublished: Like a handful of other neuronal types in the brain, cholinergic neurons (CNs) in the pedunculopontine nucleus (PPN) are lost in the course of Parkinson’s disease (PD). PPN CNs have a distinctive physiological phenotype that shares some, but not all, of the features of other neurons that are selectively vulnerable in PD. View original preprint.
Functional characterization of ATP13A2 variants associated with distinct neurodegenerative disorders
By savannah onPreprint: Loss-of-function variants in ATP13A2 are causative for Kufor-Rakeb syndrome (KRS, a recessive juvenile-onset parkinsonism with dementia) and also identified in early-onset PD (EOPD) and hereditary spastic paraplegia (HSP). The ALS and MSA variants presented a subtle functional defect, questioning whether these heterozygous variants are pathogenic and ATP13A2 dysfunction may cause MSA or ALS.
Manual Tracking/measure_ECM.ijm
By savannah onA script to generate circular ROIs in each frame of a timelapse, using a previously generated track as reference. Used to measure fluorescence intensity (i.e., extracellular matrix content) along a microglia track. To be used as a continuation of 2_tracking.ijm.
Manual Tracking/tracking.ijm
By savannah onA script to store in ImageJ’s ROImanager the tracks generated using Manual Tracking plugin, for later use (drawing tracks, frame-by-frame analysis). To be used as a continuation of 1_image_prep.ijm.
Manual Tracking/image_prep.ijm
By savannah onA script to register 2-channel timelapse images using HyperStackReg, using only 1 of the channels to compute transformation. The script also enhances contrast and prepares the image for manual tracking.
Free-floating mouse brain immunohistochemistry
By savannah onThis protocol enables immunohistochemical staining of murine tissue with superior penetration of the tissue by the reagents due to the free-floating approach. Watch an interview about this protocol with Jonathan Breiter, MSc.
