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DOI: 10.1369/jhc.3A6244.2004
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Journal of Histochemistry and Cytochemistry
Volume 52 (9): 1177-1189, 2004
Copyright ©The Histochemical Society, Inc.

Tyrosine Hydroxylase Induction by Basic Fibroblast Growth Factor and Cyclic AMP Analogs in Striatal Neural Stem Cells : Role of ERK1/ERK2 Mitogen-activated Protein Kinase and Protein Kinase C

Miguel A. López-Toledano1, Carolina Redondo, Maria V.T. Lobo, Diana Reimers, Antonio S. Herranz, Carlos L. Paíno and Eulalia Bazán

Servicios de Neurobiología (MAL-T,CR,MVTL,ASH,CLP,EB) and Histología (DR), Departamento de Investigación, Hospital Ramón y Cajal, Madrid, Spain; and Departamento de Biología Celular y Genética (MVTL), Universidad de Alcalá, Madrid, Spain

Correspondence to: Dr. Eulalia Bazán, Servicio de Neurobiologia-Investigación, Hospital Ramón y Cajal, Carretera de Colmenar Viejo, Km 9.1, 28034 Madrid, Spain. E-mail: eulalia.bazan{at}hrc.es

Neural stem cells (NSC) with self-renewal and multilineage potential are considered good candidates for cell replacement of damaged nervous tissue. In vitro experimental conditions can differentiate these cells into specific neuronal phenotypes. In the present study, we describe the combined effect of basic fibroblast growth factor (bFGF) and dibutyryladenosine 3',5'-cyclic monophosphate (dbcAMP) on the differentiation of fetal rat striatal NSC into tyrosine hydroxylase-positive cells. Tyrosine hydroxylase induction was accompanied by the activation of ERK1/ERK2 mitogen-activated protein kinase and was inhibited by the ERK1/ERK2 pathway blocker PD98059, suggesting that ERK activation may be important for this process. In addition, protein kinase C (PKC) was shown to be required for tyrosine hydroxylase protein expression. The inhibition of PKC by staurosporin, as well as its downregulation, decreased the ability of bFGF+dbcAMP to generate tyrosine hydroxylase-positive cells. Moreover, the PKC activator phorbol 12-myristate 13-acetate (PMA) together with bFGF and dbcAMP led to a significant increase in phospho-ERK1/ERK2 levels, and the percentage of ß-tubulin III-positive cells that expressed tyrosine hydroxylase increased by 3.5-fold. PMA also promoted the phosphorylation of the cyclic AMP response element binding protein that might contribute to the increase in tyrosine hydroxylase-positive cells observed in bFGF+dbcAMP+PMA-treated cultures. From these results, we conclude that the manipulation in vitro of NSC from rat fetal striatum with bFGF, cyclic AMP analogs, and PKC activators promotes the generation of tyrosine hydroxylase-positive neurons. (J Histochem Cytochem 52:1177–1189, 2004)

Key Words: neurospheres • rat • dopamine • trophic factors • second messengers • signal transduction


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