Merck
CN
  • Recording membrane potential in adult neural stem cells as readout for stem cell activation following neural circuit stimulation in mouse hippocampal slices.

Recording membrane potential in adult neural stem cells as readout for stem cell activation following neural circuit stimulation in mouse hippocampal slices.

STAR protocols (2021-02-23)
Brent Asrican, Juan Song
ABSTRACT

The transition from quiescence to activation in radial neural stem cells (rNSCs) is a key first step in the process of adult neurogenesis, which can be monitored in the context of circuit activation in live tissue by whole-cell patch-clamp recording of membrane potentials of rNSCs in acute brain slices. However, membrane recordings in small non-neuronal cells such as rNSCs can be challenging. Here, we describe the preparation of materials, recording and stimulation protocols, and analyses necessary to evaluate the efficacy of activity-dependent control of rNSC behavior. For complete details on the use and execution of this protocol, please refer to Asrican et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(+)-Sodium L-ascorbate, crystalline, ≥98%
Sigma-Aldrich
Sodium pyruvate, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99%
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Guanosine 5′-triphosphate sodium salt hydrate, ≥95% (HPLC), powder
Sigma-Aldrich
N-Methyl-D-glucamine, 99.0-100.5% (titration)
Sigma-Aldrich
Adenosine 5′-triphosphate disodium salt hydrate, microbial, BioReagent, suitable for cell culture, ≥99% (HPLC)
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioUltra, for molecular biology, ≥99.0% (T)