INTRODUCTION OF THE GLUTAMATE-RECEPTOR SUBUNIT-1 INTO MOTOR-NEURONS IN-VITRO AND IN-VIVO USING A RECOMBINANT HERPES-SIMPLEX VIRUS

We developed and characterized a recombinant herpes simplex virus vect or and used it to introduce the complementary DNA encoding glutamate r eceptor subunit 1 flip into postmitotic motor neurons. Infection of pu rified motor neurons in vitro with this vector resulted in selective, high-level expression of glutamate receptor subunit 1 immunoreactivity in nearly 100% of the neurons. Patch-clamp experiments demonstrated t hat the protein product of the glutamate receptor subunit 1 flip trans gene assembles into functional pha-amino-3-hydroxy-5-methyl-4-isoxazol epropionate (AMPA) receptor channels. Herpes simplex virus-glutamate r eceptor subunit 1 flip was introduced into spinal cord cells by direct injection into the Ventral horn and selectively into motor neurons by sciatic nerve injection. High levels of expression were sustained for at least one week and were accompanied by changes in the ionic permea bility of AMPA receptors in transgene-expressing neurons. Throughout t he first week of infection, there was little evidence for toxicity. He rpes simplex virus provides a versatile tool for manipulating the glut amate receptor phenotype of postmitotic neurons and will permit study of the role of individual glutamate receptor subunits in neuronal phys iology and pathophysiology. (C) 1997 IBRO. Published by Elsevier Scien ce Ltd.