Detection of mRNA for proteins involved in retinol metabolism in iris pigment epithelium

Graefes Arch Clin Exp Ophthalmol. 1999 Dec;237(12):1046-51. doi: 10.1007/s004170050343.

Abstract

Background: To investigate in iris pigment epithelium (IPE) the expression of mRNA for proteins involved in retinol metabolism we used a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) technique.

Methods: RNA was prepared from freshly isolated bovine IPE and retinal pigment epithelium (RPE) cells and reverse transcribed. The expression of mRNA for cellular retinaldehyde binding protein (CRALBP), p63 (RPE63), the presumed retinal pigment epithelial membrane receptor for retinoids, and 11-cis-dehydrogenase (11cisRDH) was determined by RT-PCR using specific primers. Semi-quantitative expression data were obtained by using a series of fivefold dilution of each cDNA with a fixed number of PCR cycles.

Results: Bovine IPE and RPE cells express mRNA for CRALBP, 11cisRDH, and RPE63. The mRNA expression for CRALBP and 11cisRDH is high and equal in both cell types. However, RPE63 mRNA expression in IPE cells is relatively low compared with the expression in RPE cells.

Conclusions: The presence of mRNA for CRALBP, RPE63, and 11cisRDH suggests that IPE cells may be able to metabolize retinol.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Oxidoreductases / biosynthesis
  • Aldehyde Oxidoreductases / genetics
  • Animals
  • Antibodies, Monoclonal
  • Carrier Proteins / biosynthesis
  • Carrier Proteins / genetics
  • Cattle
  • DNA Primers / chemistry
  • Eye Proteins / biosynthesis
  • Eye Proteins / genetics*
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression
  • Glyceraldehyde-3-Phosphate Dehydrogenases / biosynthesis
  • Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
  • Iris / cytology
  • Iris / metabolism*
  • Membrane Proteins*
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / genetics
  • Pigment Epithelium of Eye / cytology
  • Pigment Epithelium of Eye / metabolism*
  • RNA, Messenger / biosynthesis*
  • Retinal Dehydrogenase
  • Retinaldehyde / biosynthesis
  • Retinaldehyde / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Trans-Activators*
  • Vitamin A / metabolism*
  • beta 2-Microglobulin / biosynthesis
  • beta 2-Microglobulin / genetics

Substances

  • 11-cis-retinal-binding protein
  • Antibodies, Monoclonal
  • CKAP4 protein, human
  • Carrier Proteins
  • DNA Primers
  • Eye Proteins
  • Membrane Proteins
  • Phosphoproteins
  • RNA, Messenger
  • Trans-Activators
  • beta 2-Microglobulin
  • Vitamin A
  • Aldehyde Oxidoreductases
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • Retinal Dehydrogenase
  • Retinaldehyde