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 for gene transfer. To circumvent this, we have used a defective herpes simplex 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 vIE1GT was readily immunoprecipitated from membrane fractions of vIE1GT-infected Vero cells. By using indirect double immunofluorescence techniques, vIE1GT was shown to be capable of enhancing GT expression in cultured hippocampal neurons and glia. Glucose transport in such vIE1GT-infected cultures was increased approximately 2-fold relative to controls. The efficacy of this system in vivo was then tested by microinjection of vIE1GT into adult rat hippocampus. When examined 2 days later, GT expression from vIE1GT was demonstrated in hippocampal neurons by in situ hybridization; a small but significant increase in glucose transport was detected in tissue immediately surrounding the injection site by 2-deoxy[14C]glucose uptake and autoradiography. Such injections did not cause marked cytopathology. Thus, this approach can be used to alter central nervous system physiology in vitro and in vivo.
Department of Biological Sciences, Stanford University, CA 94305.
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D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000502:physiology*; D002478:Cells, Cultured; D003673:Defective Viruses / Q000235:genetics; D005333:Fetus; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005822:Genetic Vectors; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006624:Hippocampus / Q000502:physiology*; D017403:In Situ Hybridization; D008869:Microtubule-Associated Proteins / Q000032:analysis; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010957:Plasmids; D051381:Rats; D017207:Rats, Sprague-Dawley; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014709:Vero Cells
D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D009004:Monosaccharide Transport Proteins