title: Preferential adenovirus-mediated transduction of cells at the sites of laser photocoagulation in the rat eye.
abstract: OBJECTIVE
  This study aimed to investigate the feasibility of recombinant adenovirus-mediated gene transfer into cells implicated in the development of choroidal neovascularization (CNV).
  
  
  METHODS
  A rat model of CNV which used laser photocoagulation was developed. Gene delivery into the laser spots was investigated following subretinal injection of recombinant adenoviruses, AdRSVlacZ, AdCMVlacZ or AdCMVgfp. Immunohistochemical analysis was performed using a proliferating cell nuclear antigen antibody and a cytokeratin-specific antibody to identify the cell types transduced by the recombinant adenoviruses.
  
  
  RESULTS
  At 7 days post-injection, lacZ expression was detected in 51.6 +/- 13.2% and 71.2 +/- 19.3% of laser spots in AdRSVlacZ- and AdCMVlacZ-injected eyes, respectively. By 28 days post-injection, lacZ expression was only present in AdCMVlacZ-injected eyes. In vivo fundus fluorescent photography of AdCMVgfp-injected eyes detected gfp expression in 79.9 +/- 12.7% and 35.6% +/- 19.7% of laser spots at 4 and 7 days post-injection, respectively. Although fundus fluorescent photography did not detect the gfp signal at 10 days post-injection, fluorescent microscopy revealed a gfp signal in 81.3 +/- 6.0% of laser spots. Immunohistochemical analysis detected retinal pigment epithelial (RPE) cells as the most predominant proliferating cell type in the laser spots, although several other proliferating cell types were also identified. X-gal staining showed that the majority of transduced cells were those present in the laser spots.
  
  
  CONCLUSIONS
  It is proposed that following laser photocoagulation, proliferating RPE cells are susceptible to adenovirus-mediated gene delivery and that they may be suitable targets for the delivery of antiangiogenic factors by recombinant adenoviruses in order to inhibit developing CNV membranes.
authors.affiliation: The University of Western Australia, Centre for Ophthalmology and Visual Science, Australia.
authors.forename: C M
authors.initials: CM
authors.lastname: Lai
authors.forename: W Y
authors.initials: WY
authors.lastname: Shen
authors.forename: I J
authors.initials: IJ
authors.lastname: Constable
authors.forename: P E
authors.initials: PE
authors.lastname: Rakoczy
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002455:Cell Division; D020256:Choroidal Neovascularization / Q000517:prevention & control; D005240:Feasibility Studies; D007150:Immunohistochemistry; D007633:Keratins / Q000378:metabolism; D007763:Lac Operon / Q000502:physiology; D017075:Laser Coagulation; D009799:Ocular Physiological Phenomena; D013508:Ophthalmologic Surgical Procedures; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000502:physiology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D051381:Rats; D011995:Recombination, Genetic; D014162:Transfection
chemical_list: D018809:Proliferating Cell Nuclear Antigen; D007633:Keratins