ALTERING CENTRAL-NERVOUS-SYSTEM PHYSIOLOGY WITH A DEFECTIVE HERPES-SIMPLEX VIRUS VECTOR EXPRESSING THE GLUCOSE TRANSPORTER GENE

Because of their postmitotic nature, neurons are difficult subjects fo r gene transfer. To circumvent this, we have used a defective herpes s implex virus vector to overexpress the rat brain glucose transporter ( GT) gene under the control of the human cytomegalovirus ie1 promoter. This vector, designated vIE1GT, was propagated using a herpes simplex virus type 1 temperature-sensitive mutant, ts756. GT expressed from vI E1GT was readily immunoprecipitated from membrane fractions of vIE1GT- infected Vero cells. By using indirect double immunofluorescence techn iques, vIE1GT was shown to be capable of enhancing GT expression in cu ltured hippocampal neurons and glia. Glucose transport in such vIE1GT- infected cultures was increased almost-equal-to 2-fold relative to con trols. The efficacy of this system in vivo was then tested by microinj ection of vIE1GT into adult rat hippocampus. When examined 2 days late r, GT expression from vIE1GT was demonstrated in hippocampal neurons b y in situ hybridization; a small but significant increase in glucose t ransport was detected in tissue immediately surrounding the injection site by 2-deoxy[C-14]glucose uptake and autoradiography. Such injectio ns did not cause marked cytopathology. Thus, this approach can be used to alter central nervous system physiology in vitro and in vivo.