Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid
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 i...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2004-01-01
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| Series: | Neural Plasticity |
| Online Access: | http://dx.doi.org/10.1155/NP.2004.45 |
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| author | Floriana Volpicelli Claudia Consales Massimiliano Caiazzo Luca Colucci-D'Amato Carla Perrone-Capano Umberto di Porzio |
| author_facet | Floriana Volpicelli Claudia Consales Massimiliano Caiazzo Luca Colucci-D'Amato Carla Perrone-Capano Umberto di Porzio |
| author_sort | Floriana Volpicelli |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-8b2f11877a2d4fbda2629454b6dde78f |
| institution | OA Journals |
| issn | 2090-5904 1687-5443 |
| language | English |
| publishDate | 2004-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Neural Plasticity |
| spelling | doaj-art-8b2f11877a2d4fbda2629454b6dde78f2025-08-20T02:09:25ZengWileyNeural Plasticity2090-59041687-54432004-01-01111-2455710.1155/NP.2004.45Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic AcidFloriana Volpicelli0Claudia Consales1Massimiliano Caiazzo2Luca Colucci-D'Amato3Carla Perrone-Capano4Umberto di Porzio5Institute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyInstitute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyInstitute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyInstitute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyInstitute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyInstitute of Genetics and Biophysics “A. Buzzati Traverso ”, Via P. Castellino 111, CNR, Naples 80135, ItalyWe 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.http://dx.doi.org/10.1155/NP.2004.45 |
| spellingShingle | Floriana Volpicelli Claudia Consales Massimiliano Caiazzo Luca Colucci-D'Amato Carla Perrone-Capano Umberto di Porzio Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid Neural Plasticity |
| title | Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid |
| title_full | Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid |
| title_fullStr | Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid |
| title_full_unstemmed | Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid |
| title_short | Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid |
| title_sort | enhancement of dopaminergic differentiation in proliferating midbrain neuroblasts by sonic hedgehog and ascorbic acid |
| url | http://dx.doi.org/10.1155/NP.2004.45 |
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