Peripheral nerve pericytes originating from the blood-nerve barrier expresses tight junctional molecules and transporters as barrier-forming cells.
The objective of this study was to establish pure blood-nerve barrier (BNB)-derived peripheral nerve pericyte cell lines and to investigate their unique properties as barrier-forming cells. We isolated peripheral nerve, brain, and lung pericytes from transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene. These cell lines expressed several pericyte markers such as alpha-smooth muscle actin, NG2, osteopontin, and desmin, whereas they did not express endothelial cell markers such as vWF and PECAM. In addition, these cell lines expressed several tight junction molecules such as occludin, claudin-12, ZO-1, and ZO-2. In particular, the expression of occludin was detected in peripheral nerve and brain pericytes, although it was not detected in lung pericytes by a Western blot analysis. An immunocytochemical analysis confirmed that occludin and ZO-1 were localized at the cell-cell boundaries among the pericytes. Brain and peripheral nerve pericytes also showed significantly higher trans-pericyte electrical resistance values and lower inulin clearances than lung pericytes. We considered that occludin localized at the cell-cell boundaries among the pericytes might mechanically stabilize the microvessels of the BNB and the blood-brain barrier. Furthermore, we also showed that these cell lines expressed many barrier-related transporters. ABCG2, p-gp, MRP-1, and Glut-1 were detected by a Western blot analysis and were observed in the cytoplasm and outer membrane by an immunocytochemical analysis. These transporters on pericytes might facilitate the peripheral nerve-to-blood efflux and blood-to-peripheral nerve influx transport of substrates in cooperation with those on endothelial cells in order to maintain peripheral nerve homeostasis.
Department of Neurology and Clinical Neuroscience, Yamaguchi University Graduate School of Medicine, Ube, Japan.
Fumitaka
F
Shimizu
Yasuteru
Y
Sano
Toshihiko
T
Maeda
Masa-Aki
MA
Abe
Hiroto
H
Nakayama
Ri-Ichi
R
Takahashi
Masatsugu
M
Ueda
Sumio
S
Ohtsuki
Tetsuya
T
Terasaki
Masuo
M
Obinata
Takashi
T
Kanda
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis; D049428:Blood-Nerve Barrier / Q000166:cytology / Q000378:metabolism*; D015153:Blotting, Western; D001921:Brain / Q000098:blood supply; D002199:Capillary Permeability; D002460:Cell Line; D004351:Drug Resistance; D017097:Electric Impedance; D005260:Female; D007150:Immunohistochemistry; D007444:Inulin / Q000378:metabolism; D008168:Lung / Q000098:blood supply; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D020286:Pericytes / Q000187:drug effects / Q000378:metabolism*; D011691:Puromycin / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012584:Sciatic Nerve / Q000098:blood supply*; D019108:Tight Junctions / Q000378:metabolism*; D013997:Time Factors; D015854:Up-Regulation
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000952:Antigens, Polyomavirus Transforming; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D012333:RNA, Messenger; D011691:Puromycin; D007444:Inulin