Retinal selectivity of gene expression in rat retinal versus brain capillary endothelial cell lines by differential display analysis.
OBJECTIVE
The retina is a neural tissue especially differentiated for vision and, thus, the inner blood-retinal barrier (inner BRB) specific molecules may play an essential role in maintaining neural functions in the retina. The purpose of the present study was to identify selectively expressed genes at the inner blood-retinal barrier compared with the blood-brain barrier (BBB).


METHODS
A comparison of expressed genes between conditionally immortalized rat retinal (TR-iBRB) cell lines and brain capillary endothelial (TR-BBB) cell lines was performed using mRNA differential display analysis and quantitative real time PCR analysis. The rat M-cadherin gene was cloned by performing 5' RACE, and its protein expression was detected by immunoblot analysis.


RESULTS
Eight clones were identified as highly expressed genes in TR-iBRB cells including GATA-binding protein-3 (GATA-3), cytosolic branched chain amino transferase (BCATc), and M-cadherin (cadherin-15). The rat M-cadherin gene was cloned from TR-iBRB cells, for the first time, and has >86% amino acid sequence identity to the previously cloned mammalian M-cadherins. Rat M-cadherin expression in TR-iBRB cells was much greater than that in TR-BBB cells as far as mRNA and protein levels were concerned.


CONCLUSIONS
M-cadherin, GATA-3, and BCATc are highly expressed in TR-iBRB cells compared with TR-BBB cells and may indeed be involved in unique functions at the inner BRB.
Tomi
Masatoshi
M
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Toyama, Japan.
Abukawa
Hayato
H
Nagai
Yoko
Y
Hata
Toshio
T
Takanaga
Hitomi
H
Ohtsuki
Sumio
S
Terasaki
Tetsuya
T
Hosoya
Ken-ichi
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001812:Blood-Brain Barrier / Q000502:physiology; D001813:Blood-Retinal Barrier / Q000502:physiology; D001921:Brain / Q000098:blood supply*; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D002196:Capillaries; D002460:Cell Line; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D050990:GATA3 Transcription Factor; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012171:Retinal Vessels / Q000166:cytology*; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D000637:Transaminases / Q000235:genetics* / Q000378:metabolism
D015820:Cadherins; D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; D012333:RNA, Messenger; D015534:Trans-Activators; C070896:M-cadherin; C490589:Bcat1 protein, rat; D000637:Transaminases