Adenovirus-mediated gene transfer in neurons: construction and characterization of a vector for heterologous expression of the axonal cell adhesion molecule axonin-1
| Autor: | Stephan Kunz, Urs Ziegler, Peter Sonderegger, Beat Kunz, Roman J. Giger, Wim T. J. M. C. Hermens |
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| Přispěvatelé: | University of Zurich, Sonderegger, P |
| Rok vydání: | 1997 |
| Předmět: |
DNA
Complementary animal structures Transcription Genetic Neurite Cell Adhesion Molecules Neuronal Recombinant Fusion Proteins Genetic Vectors Cytomegalovirus Genome Viral Biology medicine.disease_cause Injections Viral vector Stereotaxic Techniques Mice Ganglia Spinal Contactin 2 Tumor Cells Cultured 10019 Department of Biochemistry medicine Animals Humans Rats Wistar Axon Promoter Regions Genetic Neurons Cell adhesion molecule Adenoviruses Human General Neuroscience Genetic transfer Defective Viruses 2800 General Neuroscience Rats Cell biology Adenoviridae Facial Nerve Enhancer Elements Genetic medicine.anatomical_structure Gene Expression Regulation Dentate Gyrus 570 Life sciences biology Neuron Heterologous expression Neuroscience HeLa Cells |
| Zdroj: | Journal of Neuroscience Methods. 71:99-111 |
| ISSN: | 0165-0270 |
| DOI: | 10.1016/s0165-0270(96)00130-6 |
| Popis: | By homologous recombination, a first-generation adenovirus-based gene transfer vector, AdCMVax-1, was constructed as a means of manipulating the expression level of the axonal cell adhesion molecule axonin-1 in neurons and glial cells. AdCMVax-1 harbours the entire coding region of the chicken axonin-1 cDNA under the transcriptional control of the Cytomegalovirus enhancer/promoter in the early-region 1 of the viral genome. Characterization of AdCMVax-1 in vitro revealed highly efficient gene transfer and expression of recombinant axonin-1 in neurons and glial cells of dissociated rat dorsal root ganglia. Similar to its native counterpart, virus-derived axonin-1 was detected on the cell body, neurites, and growth cones of transduced neurons, occurred in a secreted and membrane-associated form, and could be cleaved from the membrane with phosphatidylinositol-specific phospholipase C. Functional characterization of recombinant axonin-1 revealed the same binding properties as previously reported for native axonin-1 isolated from the vitreous fluid of chicken embryos. In vivo gene transfer was studied by stereotactic injection of AdCMVax-1 in the dentate gyrus of the hippocampus and the facial nucleus in the brainstem of adult Wistar rats and revealed high level expression of recombinant axonin-1 in a subset of hippocampal neurons and motor neurons in the facial nucleus. |
| Databáze: | OpenAIRE |
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