title: Generation of subtype-specific neurons from postnatal astroglia of the mouse cerebral cortex.
abstract: Instructing glial cells to generate neurons may prove to be a strategy to replace neurons that have degenerated. Here, we describe a robust protocol for the efficient in vitro conversion of postnatal astroglia from the mouse cerebral cortex into functional, synapse-forming neurons. This protocol involves two steps: (i) expansion of astroglial cells (7 d) and (ii) astroglia-to-neuron conversion induced by persistent and strong retroviral expression of Neurog2 (encoding neurogenin-2) or Mash1 (also referred to as achaete-scute complex homolog 1 or Ascl1) and/or distal-less homeobox 2 (Dlx2) for generation of glutamatergic or GABAergic neurons, respectively (7-21 d for different degrees of maturity). Our protocol of astroglia-to-neuron conversion by a single neurogenic transcription factor provides a stringent experimental system to study the specification of a selective neuronal subtype, thus offering an alternative to the use of embryonic or neural stem cells. Moreover, it can be a useful model for studies of lineage conversion from non-neuronal cells, with potential for brain regenerative medicine.
authors.affiliation: Department of Physiological Genomics, Institute of Physiology, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Christophe
authors.initials: C
authors.lastname: Heinrich
authors.forename: Sergio
authors.initials: S
authors.lastname: Gascón
authors.forename: Giacomo
authors.initials: G
authors.lastname: Masserdotti
authors.forename: Alexandra
authors.initials: A
authors.lastname: Lepier
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Sanchez
authors.forename: Tatiana
authors.initials: T
authors.lastname: Simon-Ebert
authors.forename: Timm
authors.initials: T
authors.lastname: Schroeder
authors.forename: Magdalena
authors.initials: M
authors.lastname: Götz
authors.forename: Benedikt
authors.initials: B
authors.lastname: Berninger
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000821:virology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D002540:Cerebral Cortex / Q000166:cytology*; D003470:Culture Media; D004594:Electrophysiology; D018014:Gene Transfer Techniques; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D018408:Patch-Clamp Techniques; D044968:Regenerative Medicine / Q000379:methods; D012190:Retroviridae / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D003470:Culture Media; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D014157:Transcription Factors; C499317:Ascl1 protein, mouse; C088334:Distal-less homeobox proteins; C102938:Neurog2 protein, mouse