Output Catalog
ASAP is committed to accelerating the pace of discovery and informing a path to a cure for Parkinson’s disease through collaboration, research-enabling resources, and data sharing. We created this catalog to showcase the research outputs and tools developed by ASAP-funded programs.
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Output Type
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Program
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CRN Team Name
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Theme
in vivo TurboID
Protocol uses iBioID with Lamp1-TurboID in astrocytes to study endo-/lysosomal protein interactions in mice. AAV delivery allows selective biotinylation for proteome mapping, offering insights into astrocyte signaling in vivo.
Postnatal astrocyte labeling by electroporation
Here, we provide a step-by-step guide to perform Postnatal Astrocyte Labeling by Electroporation (PALE), detailing the preparation, electroporation procedure, and post-electroporation analysis.
RT-qPCR (primary cells)
Protocol details qPCR method for Atp13a4 expression in astrocytes and neurons using Fast SYBR Green Master Mix in a 96-well format. Normalization to 18S gene ensures accurate quantification for reliable gene expression analysis.
RNA isolation and cDNA synthesis
This protocol details the isolation of RNA, purification, and subsequent cDNA synthesis for downstream applications such as qPCR.
Lentiviral vector production and primary astrocyte transduction
Protocol for lentivirus production & transduction into astrocytes is outlined, allowing efficient transduction & stable selection. Facilitates research on primary gene function in astrocytes.
Astrocyte morphology assessment in astrocyte-neuron co-cultures
This protocol outlines the process for analyzing astrocyte morphology in astrocyte-neuron co-cultures using immunostaining and Sholl analysis.
Multiplexed immunofluorescence and RNA-FISH
Protocol detects Atp13a4 mRNA in astrocytes of visual cortex using immunofluorescence and RNA-FISH. Sections from Aldh1L1-EGFP mice are stained, hybridized with probes, and imaged for spatial analysis of Atp13a4 expression in postnatal development.
Glia-Free Cortical Neuronal Feeding Schedule – Neuronal Morphology
This protocol outlines the treatment of cortical neurons with astrocyte-conditioned medium (ACM) or spermidine and subsequent analysis of dendrite length.
Astrocyte Isolation and Transfection for Astrocytes/Neuron co-culture assay
Protocol for isolating rat cortical astrocytes and producing astrocyte conditioned media for synaptogenesis assays.
Synapse staining – mouse brain sections
This protocol details a method to visualize and quantify excitatory synapses using mouse brain sections.
Isolation and culture of primary mouse astrocytes and neurons
This protocol details the methodology to establish primary murine astrocyte and neuron cultures.
In vivo Electrophysiology Protocol
Protocol outlines method for collecting extracellular single unit dopamine recordings.
DART Infusion Protocol
The protocol outlines the procedure for DART (drug acutely restricted by tethering) intracranial infusion in mice
FibPho: Analysis Protocol
Protocol outlines fiber photometry data analysis.
Fiber Photometry Protocol
Protocol for fiber photometry collection from VTA dopamine neurons
