title: In Vivo Fluorescence Retinal Imaging Following AAV2-Mediated Gene Delivery in the Rat Retina.
abstract: The purpose of this study was to evaluate longitudinal gene expression patterns by retinal imaging using a modified custom-built confocal laser-scanning microscope in experimental rats after intravitreal injection of recombinant adeno-associated virus 2 (rAAV2-green fluorescent protein [GFP]).
  
  
  
  Ten 9-week-old Wistar rats were divided into two groups: experimental group (group 1) that received a rAAV2-GFP intravitreal injection and control group (group 2) that received a vehicle. After anesthesia using a Zoletil intraperitoneal injection, 8 μL rAAV2-GFP in group 1 or vehicle in group 2 was injected intravitreally using a 33-G Hamilton syringe. In vivo fluorescence retinal images were acquired under anesthesia at 2, 4, 6, and 13 days after rAAV or vehicle delivery.
  
  
  
  Differences in GFP fluorescence were identified starting from day 2 after the intravitreal injection of rAAV2-GFP in group 1. Between days 4 and 6, the intensity and area of fluorescence in the retina began to increase and peaked at day 13. Based on the pattern of GFP expression, the axon of the nerve fiber layer ganglion cell was identified. In group 2, eyes treated with the vehicle showed a small amount of autofluorescence in a limited area for up to 2 weeks, with no increase in intensity during this period.
  
  
  
  In vivo retinal imaging confirmed gene expression within 2 weeks after the intravitreal injection of rAAV2-GFP. Gene transfer and expression in the rat retina occurs quickly in 2 days and appears to peak within 2 weeks of gene delivery. In vivo retinal imaging may be a useful noninvasive tool to continuously monitor gene expression in the retina over time.
authors.affiliation: Department of Ophthalmology, Asan Medical Center, University of Ulsan, College of Medicine, Seoul, Korea.
authors.forename: Joo Yong
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea 3KI for Health Science and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea.
authors.forename: Yoonha
authors.initials: Y
authors.lastname: Hwang
authors.affiliation: Department of Microbiology, University of Ulsan, College of Medicine, Seoul, Korea 5Cellular Dysfunction Research Center, University of Ulsan, College of Medicine, Seoul, Korea.
authors.forename: Ji Hyun
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Ophthalmology, Asan Medical Center, University of Ulsan, College of Medicine, Seoul, Korea.
authors.forename: Yu Sun
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Department of Ophthalmology, Asan Medical Center, University of Ulsan, College of Medicine, Seoul, Korea.
authors.forename: Bok Kyoung
authors.initials: BK
authors.lastname: Jung
authors.affiliation: Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea 3KI for Health Science and Technology, Korea Advanced Institute of Science and Technology, Daejeon, Korea.
authors.forename: Pilhan
authors.initials: P
authors.lastname: Kim
authors.affiliation: Department of Microbiology, University of Ulsan, College of Medicine, Seoul, Korea 5Cellular Dysfunction Research Center, University of Ulsan, College of Medicine, Seoul, Korea.
authors.forename: Heuiran
authors.initials: H
authors.lastname: Lee
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D058449:Intravitreal Injections; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D023421:Models, Animal; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism*; D012165:Retinal Ganglion Cells / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins