An opsin homologue in the retina and pigment epithelium

Invest Ophthalmol Vis Sci. 1993 Dec;34(13):3669-78.

Abstract

Purpose: The aim of this project was to investigate the retinal pigment epithelium (RPE) at the molecular level by identification of novel RPE-specific cDNAs that may encode proteins of signal transduction pathways or other proteins that are expressed preferentially in the RPE.

Methods: A bovine RPE cDNA library was constructed in bacteriophage lambda g10 using RPE-enriched poly(A)+ RNA. The library was screened by differential hybridization to bovine RPE and kidney cDNA probes.

Results: A member of the hepatahelical receptor family was identified in bovine RPE by molecular cloning. Its deduced amino acid sequence predicts a protein that has 291 amino acid residues and resembles most closely the family of visual pigments. A lysine residue, analogous to the retinaldehyde attachment site in rhodopsin, is conserved in the seventh hydrophobic segment of the novel sequence. Messenger RNA encoding the putative G protein-coupled receptor was detected by in situ hybridization in the RPE, inner nuclear layer, and specific cells of the ganglion cell layer. Immunohistochemical staining of bovine retina showed that the receptor protein is localized in Müller cells, as well as in the RPE.

Conclusions: A novel heptahelical receptor defines a distant evolutionary branch of the visual pigment tree. The selective localization of this putative receptor, its abundance in RPE and retina, and its homology to the visual pigments suggest that the function of this receptor is important in a visual process involving the RPE and Müller cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacteriophages
  • Base Sequence
  • Cattle
  • DNA / analysis
  • Decapodiformes
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / genetics
  • Gene Expression
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Pigment Epithelium of Eye / metabolism*
  • RNA, Messenger / metabolism
  • Rabbits
  • Receptors, Purinergic / chemistry
  • Receptors, Purinergic / genetics
  • Retina / metabolism*
  • Rod Opsins / chemistry
  • Rod Opsins / genetics*
  • Sequence Homology, Amino Acid

Substances

  • RNA, Messenger
  • Receptors, Purinergic
  • Rod Opsins
  • DNA
  • GTP-Binding Proteins