Transduction of the inner mouse retina using AAVrh8 and AAVrh10 via intravitreal injection.
Adeno-associated virus (AAV) is a proven, safe and effective vector for gene delivery in the retina. There are over 100 serotypes of AAV, and AAV2 through AAV9 have been evaluated in the retina. Each AAV serotype has different cell tropism and transduction efficiency. Intravitreal injections of AAV into the eye tend to transduce cells in the ganglion cell layer (GCL), while subretinal injections tend to transduce retinal pigment epithelium and photoreceptors. Efficient transduction of the inner retina beyond the GCL is not well established with the current methodologies and serotypes used to date. In this study, we compared the cellular tropism of AAVrh8 and AAVrh10 vectors encoding enhanced green fluorescent protein (EGFP) using intravitreal injections. We found that AAVrh8 largely transduced cells in the GCL and also amacrine cells in the inner nuclear layer (INL), as well as Müller and horizontal cells. Inner retinal transduction with AAVrh10 was similar to AAVrh8, but AAVrh10 appeared to also transduce bipolar cells. The transduction efficiency as measured by the intensity of EGFP signal was 3.5 fold higher in horizontal cells transduced with AAVrh10 than AAVrh8. Glial fibrillary accessory protein (GFAP) levels were increased in Müller cells in transduced areas for both serotypes. The results of this study suggest that AAVrh8 and AAVrh10 may be excellent vector candidates to deliver genetic material to the INL, particularly for amacrine and horizontal cells, however they may also cause cellular stress as shown by increased glial GFAP expression.
Giove
Thomas J
TJ
Laboratory of Visual Neurobiology, Boston University, Department of Biology, 5 Cummington St, Boston, MA 02215, USA.
Sena-Esteves
Miguel
M
Eldred
William D
WD
D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D005822:Genetic Vectors; D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007267:Injections; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism; D012160:Retina / Q000378:metabolism*; D014161:Transduction, Genetic; D056189:Viral Tropism / Q000502:physiology; D014822:Vitreous Body
D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; C485184:enhanced green fluorescent protein; C528900:glial fibrillary astrocytic protein, mouse; D049452:Green Fluorescent Proteins