Maoli Duan1, Qiang Mi. 1. Department of Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden. maoli.duan@ki.se
Abstract
CONCLUSION: The results suggest that local injection of 1 microl of lentiviral-green fluorescent protein (LV-GFP) into rat scala tympani as a lentiviral (LV) vector in the cochlea does not disseminate into the surrounding brain tissue. OBJECTIVE: To investigate whether the LV vector will spread into the cerebrospinal fluid (CSF) and affect brain tissue after local cochlear injection in an animal model. MATERIALS AND METHODS: Sixteen animals were sacrificed after cochleostomy and injection of 1 microl LV-GFP vectors with different promoters such as CAG (consisting of the cytomegalovirus immediate early enhancer and the chicken beta-actin promoter), EF-1alpha (human elongation factor 1alpha), PGK (human phosphoglycerate kinase 1) and CPPT (central polypurine tract). Eleven brain tissues were fixed in 4% paraformaldehyde at 4 degrees C, processed for cryosectioning and examined under fluorescence microscope. RESULTS: The patterns of the fluorescent signals with red and green filters were compared to identify the GFP signals in the brain tissue. GFP reporter gene expression was not detected in any examined brain region in any of the animals.
CONCLUSION: The results suggest that local injection of 1 microl of lentiviral-green fluorescent protein (LV-GFP) into rat scala tympani as a lentiviral (LV) vector in the cochlea does not disseminate into the surrounding brain tissue. OBJECTIVE: To investigate whether the LV vector will spread into the cerebrospinal fluid (CSF) and affect brain tissue after local cochlear injection in an animal model. MATERIALS AND METHODS: Sixteen animals were sacrificed after cochleostomy and injection of 1 microl LV-GFP vectors with different promoters such as CAG (consisting of the cytomegalovirus immediate early enhancer and the chicken beta-actin promoter), EF-1alpha (human elongation factor 1alpha), PGK (humanphosphoglycerate kinase 1) and CPPT (central polypurine tract). Eleven brain tissues were fixed in 4% paraformaldehyde at 4 degrees C, processed for cryosectioning and examined under fluorescence microscope. RESULTS: The patterns of the fluorescent signals with red and green filters were compared to identify the GFP signals in the brain tissue. GFP reporter gene expression was not detected in any examined brain region in any of the animals.
Authors: Francisco Santaolalla; Carlos Salvador; Agustín Martínez; Jose María Sánchez; Ana Sánchez Del Rey Journal: Neural Regen Res Date: 2013-08-25 Impact factor: 5.135