Detection of mRNA for proteins involved in retinol metabolism in iris pigment epithelium.
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.
Thumann
G
G
University of Cologne, Department of Vitreoretinal Surgery, Germany. ThumannGa@aol.com
Kociok
N
N
Bartz-Schmidt
K U
KU
Esser
P
P
Schraermeyer
U
U
Heimann
K
K
D000445:Aldehyde Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000096:biosynthesis / Q000235:genetics; D007498:Iris / Q000166:cytology / Q000378:metabolism*; D008565:Membrane Proteins; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis*; D050697:Retinal Dehydrogenase; D012172:Retinaldehyde / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators; D014801:Vitamin A / Q000378:metabolism*; D001613:beta 2-Microglobulin / Q000096:biosynthesis / Q000235:genetics
C020932:11-cis-retinal-binding protein; D000911:Antibodies, Monoclonal; C081876:CKAP4 protein, human; D002352:Carrier Proteins; D017931:DNA Primers; D005136:Eye Proteins; D008565:Membrane Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D015534:Trans-Activators; D001613:beta 2-Microglobulin; D014801:Vitamin A; D000445:Aldehyde Oxidoreductases; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D050697:Retinal Dehydrogenase; D012172:Retinaldehyde