Application of magnetically isolated rat retinal vascular endothelial cells for the determination of transporter gene expression levels at the inner blood-retinal barrier.
The purpose of the present study was to quantify transporter gene levels at the inner blood-retinal barrier (inner BRB) using a combination of magnetic isolation method for rat retinal vascular endothelial cells (RVEC) and real-time quantitative PCR analysis. The transcript levels of CD31, Tie-2, claudin-5, occludin, Jam-1, mdr1a, oatp2, and oatp14 in the RVEC fraction were more than 100-fold greater than those in the non-RVEC fraction, suggesting that these genes are predominantly expressed at the inner BRB. The transcript levels of GLUT1 and MCT1 in the RVEC fraction were the most abundant in the respective transporter family, suggesting that GLUT1 and MCT1 play a predominant role in d-glucose and monocarboxylate transport, respectively, at the inner BRB. In conclusion, application of magnetically isolated RVEC is able to determine transporter gene levels at the inner BRB thereby increasing our understanding of inner BRB functions at a molecular level.
Tomi
Masatoshi
M
Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Sugitani, Toyama, Japan.
Hosoya
Ken-ichi
K
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001813:Blood-Retinal Barrier / Q000502:physiology*; D004730:Endothelium, Vascular / Q000378:metabolism*; D005786:Gene Expression Regulation; D008280:Magnetics; D008297:Male; D026901:Membrane Transport Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012171:Retinal Vessels / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D015415:Biomarkers; D026901:Membrane Transport Proteins; D012333:RNA, Messenger