title: Functional evaluation of neural stem cell differentiation by single cell calcium imaging.
abstract: Neurogenesis in the adult mammalian brain occurs in two specific brain areas, the subventricular zone (SVZ) bordering the lateral ventricles and the subgranular zone (SGZ) of the hippocampus. Although these regions are prone to produce new neurons, cultured cells from these neurogenic niches tend to be mixed cultures, containing both neurons and glial cells. Several reports highlight the potential of the self-healing capacity of the brain following injury. Even though much knowledge has been produced on the neurogenesis itself, brain repairing strategies are still far away from patients cure. Here we review general concepts in the neurogenesis field, also addressing the methods available to study neural stem cell differentiation. A major problem faced by research groups and companies dedicated to brain regenerative medicine resides on the lack of good methods to functionally identify neural stem cell differentiation and novel drug targets. To address this issue, we developed a unique single cell calcium imaging-based method to functionally discriminate different cell types derived from SVZ neural stem cell cultures. The unique functional profile of each SVZ cell type was correlated at the single cell level with the immunodetection of specific phenotypic markers. This platform was raised on the basis of the functional response of neurons, oligodendrocytes and immature cells to depolarising agents, to thrombin and to histamine, respectively. We also outline key studies in which our new platform was extremely relevant in the context of drug discovery and development in the area of brain regenerative medicine.
authors.affiliation: Neuroprotection and Neurogenesis in Brain Repair Group, Center for Neuroscience and Cell Biology, Institute of Biochemistry, University of Coimbra, Largo Marquês de Pombal, Portugal.
authors.forename: Maria Francisca
authors.initials: MF
authors.lastname: Eiriz
authors.forename: Sofia
authors.initials: S
authors.lastname: Grade
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rosa
authors.forename: Sara
authors.initials: S
authors.lastname: Xapelli
authors.forename: Liliana
authors.initials: L
authors.lastname: Bernardino
authors.forename: Fabienne
authors.initials: F
authors.lastname: Agasse
authors.forename: João O
authors.initials: JO
authors.lastname: Malva
mesh_terms: D000818:Animals; D001921:Brain / Q000502:physiology; D020013:Calcium Signaling; D002454:Cell Differentiation; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D006801:Humans; D020547:Lateral Ventricles / Q000166:cytology / Q000502:physiology; D058953:Neural Stem Cells / Q000166:cytology* / Q000502:physiology; D055495:Neurogenesis; D012038:Regeneration; D059010:Single-Cell Analysis / Q000379:methods*