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Enhancement of dopaminergic differentiation in proliferating midbrain neuroblasts by sonic hedgehog and ascorbic acid.

TitleEnhancement of dopaminergic differentiation in proliferating midbrain neuroblasts by sonic hedgehog and ascorbic acid.
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2004
AuthorsVolpicelli, Floriana, Consales Claudia, Caiazzo Massimiliano, Colucci-D'Amato Luca, Perrone-Capano Carla, and di Porzio Umberto
JournalNeural Plast
Volume11
Issue1-2
Pagination45-57
Date Published2004
ISSN20905904
KeywordsAnimals, Ascorbic acid, cell differentiation, Cell Division, Cells, Cultured, Dopamine, Female, Fibroblast Growth Factor 2, Hedgehog Proteins, Mesencephalon, Neurons, pregnancy, Rats, Rats, Sprague-Dawley, Stem cells, Trans-Activators
Abstract

We analyzed the molecular mechanisms involved in the acquisition and maturation of dopaminergic (DA) neurons generated in vitro from rat ventral mesencephalon (MES) cells in the presence of mitogens or specific signaling molecules. The addition of basic fibroblast growth factor (bFGF) to MES cells in serum-free medium stimulates the proliferation of neuroblasts but delays DA differentiation. Recombinant Sonic hedgehog (SHH) protein increases up to three fold the number of tyrosine hydroxylase (TH)-positive cells and their differentiation, an effect abolished by anti-SHH antibodies. The expanded cultures are rich in nestin-positive neurons, glial cells are rare, all TH+ neurons are DA, and all DA and GABAergic markers analyzed are expressed. Adding ascorbic acid to bFGF/SHH-treated cultures resulted in a further five- to seven-fold enhancement of viable DA neurons. This experimental system also provides a powerful tool to generate DA neurons from single embryos. Our strategy provides an enriched source of MES DA neurons that are useful for analyzing molecular mechanisms controlling their function and for experimental regenerative approaches in DA dysfunction.

DOI10.1155/NP.2004.45
Alternate JournalNeural Plast.
Citation Key6763
PubMed ID15303305
PubMed Central IDPMC2565440