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 ch aracterization of regulatory sequences that are involved in the contro l of cell-type specificity and ligand regulation of neuronal gene expr ession in cultured neurons. We constructed recombinant adenoviruses, i ncorporating the luciferase gene under the control of different fragme nts of the rat tyrosine hydroxylase (TH) promoter. Similar results for luciferase expression were obtained in immortalized cells either by i nfection using adenoviral constructs or by transfection using conventi onal plasmid vectors. Taking advantage of adenoviral vectors, we exten ded 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 cer vical ganglia, Furthermore, using this neuronal culture model, we show ed that the same promoter region carries leukemia-inhibitory factor (L IF)-responsive element(s). Our results demonstrate that the first 800 bp of the rat TH promoter contains a functionally important core regio n for constitutive and LIF-regulated expression of TH in peripheral no radrenergic neurons. Moreover, the study validates the adenoviral vect or-based system as a new strategy for studying the regulation of neuro nal gene expression.