» Articles » PMID: 38165800

Time-lapse Imaging of Cortical Projection Neuron Migration in Mice Using Mosaic Analysis with Double Markers

Overview
Journal STAR Protoc
Publisher Cell Press
Date 2024 Jan 2
PMID 38165800
Authors
Affiliations
Soon will be listed here.
Abstract

Mosaic analysis with double markers (MADM) technology enables the sparse labeling of genetically defined neurons. We present a protocol for time-lapse imaging of cortical projection neuron migration in mice using MADM. We describe steps for the isolation, culturing, and 4D imaging of neuronal dynamics in MADM-labeled brain tissue. While this protocol is compatible with other single-cell labeling methods, the MADM approach provides a genetic platform for the functional assessment of cell-autonomous candidate gene function and the relative contribution of non-cell-autonomous effects. For complete details on the use and execution of this protocol, please refer to Hansen et al. (2022), Contreras et al. (2021), and Amberg and Hippenmeyer (2021)..

References
1.
Tinevez J, Perry N, Schindelin J, Hoopes G, Reynolds G, Laplantine E . TrackMate: An open and extensible platform for single-particle tracking. Methods. 2016; 115:80-90. DOI: 10.1016/j.ymeth.2016.09.016. View

2.
Hansen A, Hippenmeyer S . Non-Cell-Autonomous Mechanisms in Radial Projection Neuron Migration in the Developing Cerebral Cortex. Front Cell Dev Biol. 2020; 8:574382. PMC: 7545535. DOI: 10.3389/fcell.2020.574382. View

3.
Contreras X, Amberg N, Davaatseren A, Hansen A, Sonntag J, Andersen L . A genome-wide library of MADM mice for single-cell genetic mosaic analysis. Cell Rep. 2021; 35(12):109274. PMC: 8317686. DOI: 10.1016/j.celrep.2021.109274. View

4.
Hezel M, Ebrahimi F, Koch M, Dehghani F . Propidium iodide staining: a new application in fluorescence microscopy for analysis of cytoarchitecture in adult and developing rodent brain. Micron. 2012; 43(10):1031-8. DOI: 10.1016/j.micron.2012.04.006. View

5.
Hansen A, Pauler F, Riedl M, Streicher C, Heger A, Laukoter S . Tissue-Wide Effects Override Cell-Intrinsic Gene Function in Radial Neuron Migration. Oxf Open Neurosci. 2024; 1:kvac009. PMC: 10939316. DOI: 10.1093/oons/kvac009. View