Targeted in utero delivery of a retroviral vector for gene transfer in the rodent brain.
In vivo application of viral vectors for gene transfer is a commonly used tool in anatomical and functional studies, as well as in development of neuroprotective and restorative strategies for therapy. Although the most common route of administration is via direct injection into the brain parenchyma in adult animals, a number of short-term studies have been performed in the developing central nervous system. Here we investigated the long-term transgene expression following in utero delivery of a retroviral vector encoding for the green fluorescent protein (GFP) marker gene at embryonic days 14.5-17.5 using an ultrasound-guided injection system. Intraparenchymal injections of the ganglionic eminence were compared with vector delivery to the intracerebroventricular space. Injections into the ganglionic eminences resulted in a predominantly unilateral transduction localized to the forebrain, giving rise to GFP-positive (GFP+) neurons and astrocytes in the striatum, olfactory bulb, cortex and hippocampus. When the vector was injected into the lateral ventricle, on the other hand, widespread expression of GFP was seen throughout the brain. The total number of GFP+ cells in the striatum was estimated to be between 20,000 and 50,000 cells using a computerized stereological quantification tool. Phenotypic characterization of these transduced cells using confocal microscopical analysis showed that 64% were NeuN+ neurons, 14% APC+ oligodendrocytes and 15% glial cells labelled with GFAP, S100beta and Iba1, when the vector injection was performed at E14.5. Delivery into later embryos resulted in a reduction in neuronal profiles with a reciprocal increase in glial cells.
Stott
Simon R W
SR
CNS Disease Modelling Unit, Section of Neuroscience, Department of Experimental Medical Science, Lund University, Sweden. Simon.Stott@med.lu.se
Kirik
Deniz
D
D000367:Age Factors; D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000821:virology; D002452:Cell Count / Q000379:methods; D000089622:Doublecortin Domain Proteins; D004622:Embryo, Mammalian; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D005822:Genetic Vectors / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D019394:Ki-67 Antigen / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D012190:Retroviridae / Q000502:physiology*; D012794:Sialic Acids / Q000378:metabolism; D013004:Somatostatin / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods; D014599:Uterus / Q000502:physiology* / Q000601:surgery / Q000821:virology
D000089622:Doublecortin Domain Proteins; D019394:Ki-67 Antigen; D008869:Microtubule-Associated Proteins; D039842:Neural Cell Adhesion Molecule L1; D009479:Neuropeptides; D012794:Sialic Acids; C103469:polysialyl neural cell adhesion molecule; D049452:Green Fluorescent Proteins; D013004:Somatostatin; D010751:Phosphopyruvate Hydratase