Utilization of an HSV-based amplicon vector encoding the axonal marker hPLAP to follow neurite outgrowth in cultured DRG neurons.
Delivery of genes into DRG neurons by viral vectors is a powerful tool for the study of axonal outgrowth. In order to achieve efficient transfer of growth-related genes and simultaneously label neuronal processes, we have utilized the HSV-based amplicon vector system. A bicistronic expression cassette encoding the growth associated protein-43 (GAP-43) and the axonal marker human placental alkaline phosphatase (hPLAP) reporter gene under translation control of an internal ribosomal entry site was cloned into the HGCX amplicon vector. This hPLAP reporter enabled efficient labeling of neurites in both dissociated adult DRG neurons and embryonic DRG explants. Using this reporter, the effect of GAP-43 on neurite outgrowth in transduced DRG neurons could be demonstrated. HSV-based amplicon vectors can contribute to the study of axonal growth and guidance in cultured neurons.
Seijffers
Rhona
R
Neural Plasticity Research Group, Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, 149 13th Street, Room 4309, Charlestown, MA 02129, USA. rseijffers@partners.org
Woolf
Clifford J
CJ
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D005333:Fetus; D019922:GAP-43 Protein / Q000235:genetics / Q000378:metabolism; D005727:Ganglia, Spinal / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D020439:Growth Cones / Q000378:metabolism; D006801:Humans; D016501:Neurites / Q000378:metabolism*; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018139:Simplexvirus / Q000235:genetics*; D014162:Transfection / Q000379:methods
D015415:Biomarkers; D019922:GAP-43 Protein; D000469:Alkaline Phosphatase