Gene delivery to overcome astrocyte inhibition of axonal growth: An in vitro Model of the glial scar
| Autor: | Hannah M. Tuinstra, William E. Briley, Lonnie D. Shea, Melissa M. Ducommun |
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| Rok vydání: | 2012 |
| Předmět: |
animal structures
Neurite Genetic Vectors Bioengineering Gene delivery Biology Inhibitory postsynaptic potential Applied Microbiology and Biotechnology Article Cell Line Glial scar Extracellular matrix Cicatrix chemistry.chemical_compound Transformation Genetic Transduction Genetic medicine Animals Gene silencing Chondroitin Gene Silencing RNA Small Interfering Neurons Lentivirus Gene Transfer Techniques Models Theoretical Molecular biology Chondroitinases and Chondroitin Lyases Rats Cell biology medicine.anatomical_structure Chondroitin Sulfate Proteoglycans chemistry Astrocytes Biotechnology Astrocyte |
| Zdroj: | Biotechnology and Bioengineering. 110:947-957 |
| ISSN: | 0006-3592 |
| Popis: | After injury to the central nervous system, a glial scar develops that physically and biochemically inhibits axon growth. In the scar, activated astrocytes secrete inhibitory extracellular matrix, of which chondroitin sulfate proteoglycans (CSPGs) are considered the major inhibitory component. An inhibitory interface of CSPGs forms around the lesion and prevents axons from traversing the injury, and decreasing CSPGs can enhance axon growth. In this report, we established an in vitro interface model of activated astrocytes and subsequently investigated gene delivery as a means to reduce CSPG levels and enhance axon growth. In the model, a continuous interface of CSPG producing astrocytes was created with neurons seeded opposite the astrocytes, and neurite crossing, stopping, and turning were evaluated as they approached the interface. We investigated the efficacy of lentiviral delivery to degrade or prevent the synthesis of CSPGs, thereby removing CSPG inhibition of neurite growth. Lentiviral delivery of RNAi targeting two key CSPG synthesis enzymes, chondroitin polymerizing factor and chondroitin synthase-1, decreased CSPGs, and reduced inhibition by the interface. Degradation of CSPGs by lentiviral delivery of chondroitinase also resulted in less inhibition and more neurites crossing the interface. These results indicate that the interface model provides a tool to investigate interventions that reduce inhibition by CSPGs, and that gene delivery can be effective in promoting neurite growth across an interface of CSPG producing astrocytes. |
| Databáze: | OpenAIRE |
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