title: Predominant rod photoreceptor degeneration in Leber congenital amaurosis.
abstract: OBJECTIVE
  An unusual retinal vascular morphology in an enucleated eye from a patient with Leber congenital amaurosis (LCA) has been associated with a mutation in AIPL1. The AIPL1 protein is expressed in the pineal gland and retinal photoreceptors. In the retina, AIPL1 is expressed in both developing cone and rod photoreceptors, but it is restricted to rod photoreceptors in the adult human retina. Therefore, this study was conducted to determine the photoreceptor phenotype in this LCA patient to determine if photoreceptors were differentially affected.
  
  
  METHODS
  Additional genetic screening was performed and the consequences of the H82Y amino acid substitution characterized in an in vitro assay of NUB1 modulation. The morphology of the photoreceptors was examined by light and electron microscopy. Immunohistochemistry and immunofluorescent confocal microscopy was performed using a range of retinal photoreceptor markers.
  
  
  RESULTS
  Transfection of the H82Y mutant AIPL1 in SK-N-SH cells revealed a normal subcellular localization and solubility but resulted in an increased ability of AIPL1 to redistribute GFP-NUB1 to the cytoplasm and resolve NUB1 fragment inclusion formation. Morphologically, the LCA retina appeared to be cone-dominant with a single layer of cone-like cells remaining in the central retina. Photoreceptor outer segments were absent and the surviving residual inner segments were severely shortened. Severe degeneration of the LCA retina was associated with upregulation of glial fibrillary acidic protein (GFAP). No signal was detected for AIPL1, rhodopsin, or L/M and S cone opsins in the LCA retina. Double labeling with peanut agglutinin (PNA) and wheat germ agglutinin (WGA) supported a cone-dominant phenotype for the surviving photoreceptors in the LCA retina, as did double labeling for cone arrestin, and rod and cone recoverin. The cone arrestin signal was restricted to the residual photoreceptor inner segments and was not detected in the cell bodies, axons, or axon terminals of the surviving photoreceptors. Recoverin immunoreactivity was most intense in the residual photoreceptor inner segments.
  
  
  CONCLUSIONS
  The phenotype in this patient suggests that although AIPL1 is required for the development of normal rod and cone photoreceptor function, it might only be essential for rod and not cone survival in the adult.
authors.affiliation: Division of Pathology, Institute of Ophthalmology, University College London, London, United Kingdom.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: van der Spuy
authors.forename: Peter M G
authors.initials: PM
authors.lastname: Munro
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Luthert
authors.forename: Markus N
authors.initials: MN
authors.lastname: Preising
authors.forename: Toke
authors.initials: T
authors.lastname: Bek
authors.forename: Steffen
authors.initials: S
authors.lastname: Heegaard
authors.forename: Michael E
authors.initials: ME
authors.lastname: Cheetham
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000328:Adult; D000368:Aged; D001766:Blindness / Q000151:congenital* / Q000235:genetics / Q000473:pathology*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics*; D005136:Eye Proteins / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006162:Guanylate Cyclase / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D010641:Phenotype; D018807:Polymorphism, Single-Stranded Conformational; D011956:Receptors, Cell Surface / Q000235:genetics; D012162:Retinal Degeneration / Q000235:genetics / Q000473:pathology*; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism / Q000648:ultrastructure*; D012243:Rhodopsin / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D014162:Transfection; D015854:Up-Regulation; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D005136:Eye Proteins; D005904:Glial Fibrillary Acidic Protein; D006162:Guanylate Cyclase; D018398:Homeodomain Proteins; D011956:Receptors, Cell Surface; D012243:Rhodopsin; D015534:Trans-Activators; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase; C403625:AIPL1 protein, human; C109288:cone rod homeobox protein; C110715:guanylate cyclase 1