Neuron-Specific Transduction in the Rat Septohippocampal or Nigrostriatal Pathway by Recombinant Adeno-associated Virus Vectors

• Published in: Experimental neurology Vol. 150; no. 2; pp. 183 - 194
• Main Authors: Klein, Ronald L., Meyer, Edwin M., Peel, Alyson L., Zolotukhin, Sergei, Meyers, Craig, Muzyczka, Nicholas, King, Michael A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.04.1998; Elsevier
• Subjects: adeno-associated virus; Alzheimer's disease; Animals; Biological and medical sciences; Biotechnology; Brain-Derived Neurotrophic Factor - biosynthesis; Corpus Striatum - physiology; Cytomegalovirus - genetics; Dependovirus; Fundamental and applied biological sciences. Psychology; gene transfer; Gene Transfer Techniques; Genes, Immediate-Early; Genetic engineering; Genetic technics; Genetic Vectors; green fluorescent protein; Green Fluorescent Proteins; Hippocampus - physiology; Interneurons - cytology; Interneurons - physiology; Luminescent Proteins - biosynthesis; Methods. Procedures. Technologies; Neurons - cytology; Neurons - physiology; neurotrophic factors; Parkinson's disease; Phosphopyruvate Hydratase - biosynthesis; Phosphopyruvate Hydratase - genetics; Promoter Regions, Genetic; Proto-Oncogene Proteins c-myc - biosynthesis; Rats; Rats, Sprague-Dawley; Recombinant Fusion Proteins - biosynthesis; Substantia Nigra - physiology; Vectors (cloning, transfer, expression). Insertion sequences and transposons; adeno-associated virus; Parkinson's disease; gene transfer; Alzheimer's disease; green fluorescent protein; neurotrophic factors; Adenoviridae; Rat; Rodentia; Central nervous system; Genetic transfer; Virus; Neurotrophic factor; Vertebrata; Mammalia; Neuron; Animal; Transduction; Brain derived neurotrophic factor; Vector; Hippocampus; Brain (vertebrata)
• ISSN: 0014-4886; 1090-2430
• DOI: 10.1006/exnr.1997.6736

Viral vector-mediated gene transfer in brain can provide a means for gene therapy and functional studies. However, robust and persistent transgene expression in specific populations of the adult brain has been difficult to achieve. In an attempt to produce localized and persistent transduction in rat brain, we compared recombinant adeno-associated virus (rAAV) vectors incorporating either the immediate early cytomegalovirus (CMV) promoter or the neuron-specific enolase (NSE) promoter. Transduction in hippocampus resulting from the NSE promoter-containing construct was more efficient and persistent than that resulting from the CMV promoter-containing construct. Most hippocampal cells transduced with the NSE promoter had multipolar neuron morphology. Neurons with glutamatergic morphology were transduced weakly. In order to produce a local supply of neurotrophic factor to cells that degenerate under certain disease and experimental conditions, the NSE promoter was utilized to drive expression of brain-derived neurotrophic factor (BDNF) in medial septum or substantia nigra. In this construct, the NSE promoter drives dicistronic expression of BDNF and an enhanced version of green fluorescent protein (GFP). We estimated 3000–15,000 GFP-positive cells per injection of rAAV into septum or substantia nigra, a transduction ratio of 5–20 infectious virus particles per transduced cell. This frequency may be sufficient for trophic factor gene therapy as well as for investigating specific protein function in “topical (i.e., localized) transgenic” animals produced by rAAV.