Neuronal network formation from reprogrammed early postnatal rat cortical glial cells.
In the subependymal zone and the dentate gyrus of the adult brain of rodents, neural stem cells with glial properties generate new neurons in a life-long process. The identification of glial progenitors outside the neurogenic niches, oligodendrocyte precursors in the healthy brain, and reactive astrocytes after cortical injury led to the idea of using these cells as endogenous cell source for neural repair in the cerebral cortex. Recently, our group showed that proliferating astroglia from the cerebral cortex can be reprogrammed into neurons capable of action potential firing by forced expression of neurogenic fate determinants but failed to develop synapses. Here, we describe a maturation profile of cultured reprogrammed NG2+ and glial fibrillary acidic protein+ glia cells of the postnatal rat cortex that ends with the establishment of a glutamatergic neuronal network. Within 3 weeks after viral expression of the transcription factor neurogenin 2 (Ngn2), glia-derived neurons exhibit network-driven, glutamate receptor-dependent oscillations in Ca(2+) and exhibit functional pre- and postsynaptic specialization. Interestingly, the Ngn2-instructed glutamatergic network also supports the maturation of a γ-aminobutyric acid (GABA)ergic input via GABA(A) receptors in a non-cell autonomous manner. The "proof-of-principle" results imply that a single transcription factor may be sufficient to instruct a neuronal network from a glia-like cell source.
Blum
Robert
R
Physiological Genomics, Institute of Physiology, Ludwig-Maximilians University Munich, 80336 Munich, Germany.
Heinrich
Christophe
C
Sánchez
Rodrigo
R
Lepier
Alexandra
A
Gundelfinger
Eckart D
ED
Berninger
Benedikt
B
Götz
Magdalena
M
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D001640:Bicuculline / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000254:growth & development*; D000089622:Doublecortin Domain Proteins; D058785:GABA-A Receptor Agonists / Q000494:pharmacology; D058787:GABA-A Receptor Antagonists / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009118:Muscimol / Q000494:pharmacology; D009415:Nerve Net / Q000187:drug effects / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009457:Neuroglia / Q000187:drug effects / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009479:Neuropeptides / Q000378:metabolism; D018408:Patch-Clamp Techniques; D051381:Rats; D011963:Receptors, GABA-A / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D026941:Sodium Channel Blockers / Q000494:pharmacology; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D013779:Tetrodotoxin / Q000494:pharmacology; D014161:Transduction, Genetic / Q000379:methods; D014404:Tubulin / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D051792:Basic Helix-Loop-Helix Proteins; D001640:Bicuculline; D002118:Calcium; D000089622:Doublecortin Domain Proteins; D058785:GABA-A Receptor Agonists; D058787:GABA-A Receptor Antagonists; D049452:Green Fluorescent Proteins; D008869:Microtubule-Associated Proteins; D009118:Muscimol; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D011963:Receptors, GABA-A; D026941:Sodium Channel Blockers; D013779:Tetrodotoxin; D014404:Tubulin; C534807:Neurog2 protein, rat; C120531:Shank1 protein, rat; C488416:Tubb3 protein, rat