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.
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Department of Biological Sciences, Stanford University, CA 94305.
Mocarski
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Sapolsky
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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