Pregnant Swiss mice (E14.5) were anaesthetized by intramuscular injections of solution containing 117 μg of ketamine (Anesketin, Eurovet) and 0.7 μg of medetomidine hydrochloride (Domitor, Pfizer) per gram of body weight. Uterine horns were exposed, and a mixture of the required plasmids (1 to 2 μg/μl) was microinjected with the “Fast Green” dye (Sigma-Aldrich) in the lateral ventricles of embryos. Five current pulses (50-ms pulse/950-ms interval) were delivered across the head of the embryos (36 V), targeting the dorsal-medial part of the cortex. After 3 days of expression, the mothers were euthanized, the embryos were dissected, and the brains were quickly perfused with PBS and 4% paraformaldehyde (PFA). After dissection, the brains were post-fixed for 6 to 10 hours in 4% PFA at 4°C. Vibratome sections (100 m) were collected and prepared for imaging. Analysis of cell migration was performed as previously described (23) by dividing the cortical regions in 10 equal bins and by counting the number of positive neuronal nuclei in each bin. Note that also for neurons showed in Fig. 4 (E versus G), there are differences in the migration of neurons in the controls between experiments, as often observed when different control plasmids are used.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.