An adenoviral vector-based system to study neuronal gene expression: analysis of the rat tyrosine hydroxylase promoter in cultured neurons.
We validated an adenoviral vector-based system as a move toward the characterization of regulatory sequences that are involved in the control of cell-type specificity and ligand regulation of neuronal gene expression in cultured neurons. We constructed recombinant adenoviruses, incorporating the luciferase gene under the control of different fragments of the rat tyrosine hydroxylase (TH) promoter. Similar results for luciferase expression were obtained in immortalized cells either by infection using adenoviral constructs or by transfection using conventional plasmid vectors. Taking advantage of adenoviral vectors, we extended our experiments to various primary cell cultures. The first 800 bp of the TH promoter were found to be sufficient to confer a cell-type preferential activity in noradrenergic neurons of the rat superior cervical ganglia. Furthermore, using this neuronal culture model, we showed that the same promoter region carries leukemia-inhibitory factor (LIF)-responsive element(s). Our results demonstrate that the first 800 bp of the rat TH promoter contains a functionally important core region for constitutive and LIF-regulated expression of TH in peripheral noradrenergic neurons. Moreover, the study validates the adenoviral vector-based system as a new strategy for studying the regulation of neuronal gene expression.
Robert
J J
JJ
Laboratoire Mixte Rhône-Poulenc Rorer/CNRS C9923, Hôpital de la Pitié Salpêtrière, Paris, France.
Geoffroy
M C
MC
Finiels
F
F
Mallet
J
J
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D006131:Growth Inhibitors / Q000494:pharmacology; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology; D009474:Neurons / Q000502:physiology*; D016716:PC12 Cells; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051381:Rats; D014162:Transfection; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*
D006131:Growth Inhibitors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D014446:Tyrosine 3-Monooxygenase