AN ADENOVIRUS VECTOR ENCODING TYROSINE-HYDROXYLASE ACTIVITY MAY ENTERHUMAN CNS CELLS IN PRIMARY DISSOCIATED CULTURES

An adenovirus encoding tyrosine hydroxylase (TH) activity was inserted in neuronal and glial cultured cells obtained from human fetal centra l nervous system (CNS) tissue. Using a double fluorescence immunostain ing, we characterized inoculated CNS cells, with a TH antiserum and on e of the following antibodies: microtubule-associated protein (MAP(2)) and GABA for neuronal cells, vimentin (Vim) for glial cells and glial fibrillary acidic protein (GFAP) for astrocytes. The characterization of inoculated neuronal cells was established by the detection of TH-M AP(2)-stained neurons in cultures obtained from the thoracic and lumba r parts of the spinal cord where no intrinsic TH cells are described. Inoculated glial cells were characterized by the detection of TH-Vim a nd TH-GFAP-stained CNS cultured cells. We also observed GABA neurons e xpressing TH immunoreactivity which could be considered as inoculated neurons expressing the GABA phenotype. Whatever the time of inoculatio n, transfection was observed in both neuronal and glial cells, after u p to 4 months of culture. Although no precise quantitation was perform ed, the percentage of inoculation was found on microscopic inspection to be greater in glia than in neurons, as previously reported. We conc luded that a gene coding for a key neuronal enzyme can be incorporated in embryonic human glial and neuronal cells through the use of a reco mbinant adenovirus. Copyright (C) 1996 ISDN.