Elevated plus maze test for mice
By Emma Sherrell onThe elevated plus maze is a classical test to screen anxiety-like behaviors in laboratory animals.
Chemogenetic modulation of catecholaminergic neurons
By Emma Sherrell onThis protocol outlines a method for modulating noradrenergic and dopaminergic neuronal activity. Developed for longitudinal studies spanning up to four weeks, the protocol aims to assess neuromelanin's activity-dependent accumulation through daily agonist administration.
Optogenetic modulation of dopaminergic neurons
By Emma Sherrell onThis protocol describes an optogenetic activation of neuromelanine-laden dopaminergic neurons in the SNc-VTA.
Chronic optrode recordings from the Locus coeruleus (Optrode construction, implantation, and recording)
By Emma Sherrell onThis protocol covers chronic optrode construction, implantation, and recording in the Locus coeruleus (LC).
Protocol for Data Independent Acquisition – Mass spectrometry analysis – a DIA-based Organelle Proteomics
By savannah onThis protocol details a Data Independent Acquisition (DIA)-based mass spectrometry (MS) data acquisition method for proteomic profiling of the Golgi, including a description of how to construct the nano Liquid chromatography and DIA MS methods and a Data Dependent Acquisition (DDA) strategy to generate deep spectral libraries for searching the DIA data.
Neuromelanin-positive neuron density in substantia nigra image analysis
By Julia Julia Leonard onThe protocol covers the steps to measure neuromelanin-positive neuron density in substantia nigra using image analysis tools, including NZConnect (Hamamatsu), a web-based whole-slide image (WSI) viewer, Cellpose, and QuPath.
CD3 Cell density in substantia nigra and cerebral peduncle image analysis
By Julia Julia Leonard onThis protocol describes how to measure CD3 density in the substantia nigra and cerebral peduncle using haematoxylin and DAB-stained brain sections.
Hippocampal cultures, lentivirus production, vGLUT1-pHluorin imaging, and analysis
By Emma Sherrell onProcedures for mouse hippocampal cultures, lentivirus production, vGLUT1-pHluorin imaging and analysis used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."
D8 cell density in substantia nigra and cerebral peduncle image analysis
By Julia Julia Leonard onThis protocol describes how to measure CD8 density in the substantia nigra and cerebral peduncle using haematoxylin and DAB-stained brain sections.
pHluorin assays, analysis, and fluorescence microscopy
By savannah onProtocol for pHluorin assays, analysis, and fluorescence microscopy as performed in "Synapsin E-domain is essential for α-synuclein function." (https://doi.org/10.7554/eLife.89687.1)
Preparation and administration of systemic 4-AP
By Emma Sherrell onProcedure for preparation and systemic administration of 4-AP used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."
Immunoprecipitations and Western blots analysis
By savannah onProtocol for immunoprecipitation and Western blotting as performed in "Synapsin E-domain is essential for α-synuclein function." (https://doi.org/10.7554/eLife.89687.1)
Immunofluorescence and object-based colocalization analysis
By Emma Sherrell onProcedures for immunofluorescence and object-based colocalization analysis used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."
AT8 Tau pathology image analysis
By Julia Julia Leonard onThis protocol describes how to analyze AT8 tau pathology in human brain tissue (FFPE sections with IHC).
GST fusion protein production
By savannah onProtocol for GST fusion protein production as performed in "Synapsin E-domain is essential for α-synuclein function." (https://doi.org/10.7554/eLife.89687.1)
Proximity ligation assay (PLA)
By Emma Sherrell onProcedure for proximity ligation assay used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."
Pull-down assays
By savannah onProtocol for protein pull-down assays as performed in "Synapsin E-domain is essential for α-synuclein function." (https://doi.org/10.7554/eLife.89687.1)
αSyn immunohistochemistry
By Emma Sherrell onProcedure for immunohistochemistry used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."
Electron microscopy (EM) and quantification of inter-vesicular distances
By Emma Sherrell onProcedure for electron microscopy (EM) and quantification of inter-vesicular distances used in "Serine-129 phosphorylation of α-synuclein is a trigger for physiologic protein-protein interactions and synaptic function."