Skip to Content
Merck
CN
  • Combining low-density cell culture, single-cell tracking, and patch-clamp to monitor the behavior of postnatal murine cerebellar neural stem cells.

Combining low-density cell culture, single-cell tracking, and patch-clamp to monitor the behavior of postnatal murine cerebellar neural stem cells.

STAR protocols (2021-11-30)
Aida Menéndez-Méndez, Lucía Paniagua-Herranz, Luis A Olivos-Oré, Rosa Gómez-Villafuertes, Raquel Pérez-Sen, Esmerilda G Delicado, Antonio R Artalejo, Felipe Ortega
ABSTRACT

Low-density cell culture of the postnatal cerebellum, combined with live imaging and single-cell tracking, allows the behavior of postnatal cerebellar neural stem cells (NSCs) and their progeny to be monitored. Cultured cerebellar NSCs maintain their neurogenic nature giving rise, in the same relative proportions that exist in vivo, to the neuronal progeny generated by the three postnatal cerebellar neurogenic niches. This protocol describes the identification of the nature of the progeny through both post-imaging immunocytochemistry and patch-clamp recordings. For complete details on the use and execution of this protocol, please refer to Paniagua-Herranz et al. (2020b).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Guanosine 5′-triphosphate lithium salt, ~95% (HPLC), powder
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Monoclonal Anti-Glial Fibrillary Acidic Protein (GFAP) antibody produced in mouse, clone G-A-5, ascites fluid
Sigma-Aldrich
Serotonin hydrochloride, powder
Sigma-Aldrich
Monoclonal Anti-β-Tubulin III antibody produced in mouse, clone SDL.3D10, ascites fluid