Abstract
In previous studies we demonstrated that herpetic and adenoviral vectors constitute suitable tools for gene transfer into anterior pituitary (AP) cells in vitro. In order to extend these studies to in vivo systems we used the same type of vectors to transfer reporter genes at hypothalamic and pituitary level in rats. Here, we used three vectors: a) tsK/b-gal, an HSV-1 derived mutant harboring the E coli b-galactosidase gene. b) Ad.RSV.nls/b-gal, an adenoviral vector harboring the b-gal gene targeted to the nucleus. c) RAd (eGFP-TK)fus , an adenoviral vector carrying a DNA sequence coding for the green fluorescent protein (eGFP) fused to the gene for HSV1 thymidine kinase (TK). These vectors were sterotaxically injected in the hypothalamus and AP of female rats. Two days later the animals were sacrificed, the glands and hypothalami dissected and processed by the X-gal method to detect the presence of the b-gal in the tissues exposed to the appropriate vectors. In the animals injected with RAd(eGFP-TK)fus , the transgene product was identified by fluorescence microscopy. Our results showed that the herpetic vector was the most efficient in the AP, whereas the two adenoviral vectors showed a lower, although significant, transduction efficiency in the AP. At hypothalamic level, both types of vector displayed comparable transduction efficiencies. We conclude that adenoviral and herpetic vectors are potentially useful tools to implement gene therapy in the neuroendocrine system. (We acknowledge Sirex SRL for the generous loan of an Olympus BX-51 fluorescence microscope used in this study).
