title: In vivo Calcium Imaging of Evoked Calcium Waves in the Embryonic Cortex.
abstract: The dynamics of intracellular calcium fluxes are instrumental in the proliferation, differentiation, and migration of neuronal cells. Knowledge thus far of the relationship between these calcium changes and physiological processes in the developing brain has derived principally from ex vivo and in vitro experiments. Here, we present a new method to image intracellular calcium flux in the cerebral cortex of live rodent embryos, whilst attached to the dam through the umbilical cord. Using this approach we demonstrate induction of calcium waves by laser stimulation. These waves are sensitive to ATP-receptor blockade and are significantly increased by pharmacological facilitation of intracellular-calcium release. This approach is the closest to physiological conditions yet achieved for imaging of calcium in the embryonic brain and as such opens new avenues for the study of prenatal brain development. Furthermore, the developed method could open the possibilities of preclinical translational studies in embryos particularly important for developmentally related diseases such as schizophrenia and autism.
authors.affiliation: Neuroscience Center, University of Helsinki Helsinki, Finland.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Yuryev
authors.affiliation: INSERM U901, Institut de Neurobiologie de la Méditerranée (INMED), Parc Scientifique de LuminyMarseille, France; Aix-Marseille Université (AMU), UMR S901, Parc Scientifique de LuminyMarseille, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Pellegrino
authors.affiliation: School of Chemical Technology, Aalto University Espoo, Finland.
authors.forename: Ville
authors.initials: V
authors.lastname: Jokinen
authors.affiliation: Neuroscience Center, University of Helsinki Helsinki, Finland.
authors.forename: Liliia
authors.initials: L
authors.lastname: Andriichuk
authors.affiliation: Neuroscience Center, University of Helsinki Helsinki, Finland.
authors.forename: Stanislav
authors.initials: S
authors.lastname: Khirug
authors.affiliation: Neuroscience Center, University of Helsinki Helsinki, Finland.
authors.forename: Leonard
authors.initials: L
authors.lastname: Khiroug
authors.affiliation: Neuroscience Center, University of HelsinkiHelsinki, Finland; INSERM U901, Institut de Neurobiologie de la Méditerranée (INMED), Parc Scientifique de LuminyMarseille, France; Aix-Marseille Université (AMU), UMR S901, Parc Scientifique de LuminyMarseille, France.
authors.forename: Claudio
authors.initials: C
authors.lastname: Rivera
keywords: calcium imaging; cortical development; in vivo imaging; purinergic receptors; two-photon microscopy