Brain slice physiology and optogenetics
By onThe protocol outlines standard external solutions for electrophysiology experiments.
Harvesting and irradiation of mouse embryonic fibroblasts (MEFs) for hPSC cultures
By onThis protocol describes the process of harvesting and irradiating mouse embryonic fibroblasts (MEFs) to use as feeder cells for human pluripotent stem cell (hPSC) culture.
Expansion microscopy with R1441C LRRK2 MEF cells: visualization of Myc-RILPL1 and TMEM55B
By onExpansion microscopy is a super-resolution imaging technique that increases the physical distance between fluorophores from fixed cells on coverslips. Here, the authors use expansion techniques to confocal image TMEM55B and RILPL1.
H9 ES AAVS1-NGN2 FAM134B-/-; PiggyBac-Keima-RAMP4
By onES cells modified to lack FAM134B and express Keima-RAMP4 ER-phagy reporter using CRISPR/Cas9. NEUROG2 construct added to AAVS1 locus. Transfected with SERP1 and mKeima. Derived from human embryonic stem cells at blastocyst stage.
pCAG- WIPI2dK128E- cs-TEV -STREP
By onPlasmid: Mammalian expression of human WIPI2d K128E with C-terminal Strep.
Preparation and imaging of enriched Golgi from GolgiTAG-IP using Transmission Electron Microscopy
By onA protocol where Golgi, isolated from cells by GolgiTAG immunoprecipitation (IP) can be prepared and imaged using TEM. This protocol can also be used to image any organelles isolated using various organelle-IP protocols that are available.
pCMV-pOTC-3H-GFP
By onExpression of GFP fused with pOTC-3H, a leader peptide from mitochondrial matrix enzyme ornithine transcarbamylase and a thermostable three helix-bundle
Single-cell spatial transcriptomic and translatomic profiling of dopaminergic neurons in health, aging, and disease
By onThis study used two complementary approaches in a Parkinson's disease mouse model and revealed insights into dopaminergic neuron characteristics, aging markers, and potential disease genes like CASR.
Generation of CRISPR constructs
By onThis protocol details the procedure of generation of CRISPR constructs.
pU6-pegRNA-LRRK2-G2019S-3b
By onIt can be used to introduce LRRK2-G2019S mutation using prime editing.
Photometry Analysis Code
By onThis set of functions extracts GCaMP Fiber Photometry data obtained with a TDT system, performs isosbestic and exponential correction, and computes the number and amplitude of transients over time. The code was originally used for the following experiment: MFB-lesioned 6-OHDA D1-cre or A2A-cre mice were injected with a cre-dependent GCaMP6f in dorsolateral striatum and implanted with 400um optical fibers. Mice were placed in an open field arena and recorded for 30 minutes before levodopa injection, and 120 minutes after levodopa injection. The experiment includes video from 2 cameras and a TTL pulse from an Arduino indicating the time when levodopa was injected.
Image processing and 3D reconstruction
By onProtocol describing image processing and 3D reconstruction.
Sample Collection and Measurement of Serum Neurofilament Light (NfL)
By onThis protocol details the steps for the preparation of serum from human blood samples and the measurement of NfL concentration using the NF-Light Advantage kit on the HD-X Analyzer (Quanterix, Billerica, MA).
Expression and purification GST-tagged ATG13:ATG101 constructs
By onExpression and purification of GST-tagged ATG13/ATG101 constructs.
pCAG-TSF-ATG13 (363-517)
By onPlasmid: Mammalian expression of TwinStrep-Flag tagged ATG13 (363-517).
Pseudogenes limit the identification of novel common transcripts generated by their parent genes
By onGenomic sequences with high sequence similarity cause short sequencing reads to align to multiple locations, thus complicating genomic analyses. The authors investigated the impact of pseudogenes on transcriptomic analyses.
AAV injection in the nodose ganglia of mouse
By onThe gut-brain axis links the visceral organs to the medulla oblongata via the vagus nerve. Accessing to the afferent vagal pathway is important to dissect the role of cell populations in the bidirectional communication between brain and body. The jugular-nodose ganglion (JNG) complex contains the cell bodies of many heterogenous neural subpopulations responsible for sensing the physiological and pathological conditions of the thoracic and abdominal organs. However, the study of these ganglia is challenging in small animals due to size and location. Hence, in this protocol we describe a practical surgical approach to the vagal trunk, and the JNG complex in mice.
Incentivized Vigor Task
By onCode for an fMRI task in which human participants use a joystick to make speeded reaches to cued targets. This task examines incentive effects on movement, part of the Socal Kinesia and Incentivization for Parkinson's Disease (SKIP) dataset.
