title: GFAP-driven GFP expression in activated mouse Müller glial cells aligning retinal blood vessels following intravitreal injection of AAV2/6 vectors.
abstract: BACKGROUND
  Müller cell gliosis occurs in various retinal pathologies regardless of the underlying cellular defect. Because activated Müller glial cells span the entire retina and align areas of injury, they are ideal targets for therapeutic strategies, including gene therapy.
  
  
  RESULTS
  We used adeno-associated viral AAV2/6 vectors to transduce mouse retinas. The transduction pattern of AAV2/6 was investigated by studying expression of the green fluorescent protein (GFP) transgene using scanning-laser ophthalmoscopy and immuno-histochemistry. AAV2/6 vectors transduced mouse Müller glial cells aligning the retinal blood vessels. However, the transduction capacity was hindered by the inner limiting membrane (ILM) and besides Müller glial cells, several other inner retinal cell types were transduced. To obtain Müller glial cell-specific transgene expression, the cytomegalovirus (CMV) promoter was replaced by the glial fibrillary acidic protein (GFAP) promoter. Specificity and activation of the GFAP promoter was tested in a mouse model for retinal gliosis. Mice deficient for Crumbs homologue 1 (CRB1) develop gliosis after light exposure. Light exposure of Crb1(-/-) retinas transduced with AAV2/6-GFAP-GFP induced GFP expression restricted to activated Müller glial cells aligning retinal blood vessels.
  
  
  CONCLUSIONS
  Our experiments indicate that AAV2 vectors carrying the GFAP promoter are a promising tool for specific expression of transgenes in activated glial cells.
authors.affiliation: Department of Neuromedical Genetics, Netherlands Institute for Neuroscience, an Institute of the Royal Netherlands Academy of Arts and Sciences, Amsterdam, The Netherlands.
authors.forename: Wendy M
authors.initials: WM
authors.lastname: Aartsen
authors.forename: Koen W R
authors.initials: KW
authors.lastname: van Cleef
authors.forename: Lucie P
authors.initials: LP
authors.lastname: Pellissier
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoek
authors.forename: Rogier M
authors.initials: RM
authors.lastname: Vos
authors.forename: Bas
authors.initials: B
authors.lastname: Blits
authors.forename: Erich M E
authors.initials: EM
authors.lastname: Ehlert
authors.forename: Kamaljit S
authors.initials: KS
authors.lastname: Balaggan
authors.forename: Robin R
authors.initials: RR
authors.lastname: Ali
authors.forename: Joost
authors.initials: J
authors.lastname: Verhaagen
authors.forename: Jan
authors.initials: J
authors.lastname: Wijnholds
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015870:Gene Expression; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D007267:Injections; D051379:Mice; D009457:Neuroglia / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D012171:Retinal Vessels / Q000166:cytology*; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics
chemical_list: D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins
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