title: Expression of floor plate in dispersed mesencephalic cultures: role in differentiation of tyrosine hydroxylase neurons.
abstract: The availability of sufficient numbers of dopaminergic neurons for transplantation has been an important issue. Recently, it has been shown that the ventral floor plate (FP4-positive) cells and the transcription factor HNF-3beta are important in the signals that terminate proliferation and produce differentiation of the dopaminergic phenotype. In this study, dispersed mesencephalon from embryonic rats at Day 11 postcoitus (E-11), 1 day prior to the birth of TH cells, were cultured for 48 h and 1 week to evaluate TH neuronal differentiation and/or proliferation in vitro. The number of TH cells increased 14x between 48 h and 1 week in culture. In dispersed E-14 cultures, the presence of FP4 and HNF-3beta markers was demonstrated using immunohistochemistry. The majority of FP4-positive cell clusters were associated with TH neurons, suggesting that floor plate cells may have participated in TH neuron differentiation in culture. Antisense oligonucleotide probe for HNF-3beta mRNA added daily to cultured E-14 cells blocked the HNF-3beta expression, but had no effect on the FP4 or TH expression. These studies suggest a potentially important role for floor plate cells in the differentiation of TH cells, and differentiation and/or proliferation of TH cells in dispersed cultures of E-11 is demonstrated.
authors.affiliation: Department of Physiology and Biophysics, Finch University of Health Sciences/The Chicago Medical School, North Chicago, Illinois 60064, USA.
authors.forename: B R
authors.initials: BR
authors.lastname: Cohen
authors.forename: C D
authors.initials: CD
authors.lastname: Sladek
mesh_terms: D000818:Animals; D015415:Biomarkers; D002450:Cell Communication; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004298:Dopamine / Q000096:biosynthesis*; D004627:Embryonic Induction / Q000502:physiology*; D005314:Embryonic and Fetal Development; D004790:Enzyme Induction; D051542:Hepatocyte Nuclear Factor 3-beta; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology* / Q000201:enzymology; D009419:Nerve Tissue Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D009474:Neurons / Q000201:enzymology / Q000502:physiology*; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*
chemical_list: D015415:Biomarkers; D004268:DNA-Binding Proteins; C496504:Foxa2 protein, rat; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D051542:Hepatocyte Nuclear Factor 3-beta; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine