Tracking Neuronal Migration in Adult Brain Slices

Abstract

Neuronal migration is one of the fundamental processes underlying the proper assembly and function of neural circuitry. The majority of neuronal precursors are generated far away from their sites of integration and need to migrate substantial distances to reach their final destination. Neuronal migration occurs not only in the embryonic brain but also in a few regions of the adult brain such as the olfactory bulb (OB). The mechanisms orchestrating cell migration in the adult brain are, however, poorly understood, despite their clinical relevance. Here we describe a method for time-lapse imaging of cell migration in acute brain slices. This method, combined with genetic and/or pharmacological manipulations of different molecular pathways, makes it possible to determine the dynamics and molecular mechanisms of cell migration in the adult brain. In addition, time-lapse imaging in acute brain slices makes it possible to monitor cell movement in a microenvironment that closely resembles in vivo conditions and to study neuroblast displacement along with other cellular elements such as astrocytes and blood vessels.

The dynamic interplay between ATP/ADP levels and autophagy sustain neuronal migration in vivo

“…Measurement of the ATP/ADP ratio: Seven days after co-injecting lentiviral particles expressing PercevalHR or TdTomato in the SVZ, the mice were sacrificed and acute sections were prepared as described previously ( Bakhshetyan and Saghatelyan, 2015 ). Briefly, the mice were anesthetized with ketamine (100 mg/kg) and xylazine (10 mg/kg) and were perfused transcardially with modified oxygenated artificial cerebrospinal fluid (ACSF) containing (in mM): 210.3 sucrose, 3 KCl, 2 CaCl 2 .2H 2 O, 1.3 MgCl 2 .6H 2 O, 26 NaHCO 3 , 1.25 NaH 2 PO 4 .H 2 O, and 20 glucose.…”

Adult neural stem cell activation in mice is regulated by the day/night cycle and intracellular calcium dynamics

“…Fluorescence images were acquired using a confocal microscope (FV 1000; Olympus) with 60x (UPlanSApoN 60x/NA 1.42; Olympus) and 40x (UPlanSApoN 40x/NA 0.90; Olympus) oil and air immersion objectives, respectively. Time-lapse Ca 2+ imaging in acute brain slices Six to 10 weeks post-electroporation, the mice were sacrificed, and acute slices were prepared as described previously (Bakhshetyan and Saghatelyan, 2015). Briefly, the mice were anesthetized with ketamine (100 mg/kg) and xylazine (10 mg/kg) and were perfused transcardially with modified oxygenated artificial cerebrospinal fluid (ACSF) containing (in mM): 210.3 sucrose, 3 KCl, 2 CaCl 2 .2H 2 O, 1.3 MgCl 2 .6H 2 O, 26 NaHCO 3 , 1.25 NaH 2 PO 4 .H 2 O, and 20 glucose.…”

Deciphering heterogeneous populations of migrating cells based on the computational assessment of their dynamic properties

“…For the time-lapse imaging, acute brain slices were prepared as described previously (Bakhshetyan and Saghatelyan, 2015). Briefly, after anesthesia with ketamine (100 mg/kg) and xylazine (10 mg/kg), the mice were perfused transcardially with modified oxygenated artificial cerebrospinal fluid (ACSF) composed of 210.3 mM sucrose, 3 mM KCl, 2 mM CaCl 2 .2H 2 O, 1.3 mM MgCl 2 .6H 2 O, 26 mM NaHCO 3 , 1.25 mM NaH 2 PO 4 .H 2 O, and 20 mM glucose.…”