Peripheral Nerve pericytes originating from the blood–nerve barrier expresses tight junctional molecules and transporters as barrier‐forming cells

Abstract

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.

Advanced glycation end-products induce basement membrane hypertrophy in endoneurial microvessels and disrupt the blood–nerve barrier by stimulating the release of TGF-β and vascular endothelial growth factor (VEGF) by pericytes

“…We have successfully established PnMEC and peripheral nerve pericyte cell lines of human origin [15,16]. Therefore, our in vitro BNB model is considered suitable for the analysis of the production of basement-membrane-related molecules, because it is difficult to identify cells that synthesise these molecules in vivo.…”

“…Cell culture The immortalised human PnMECs, and brain and peripheral nerve pericyte cell lines were generated as described previously by our group [15,16]. The study protocol for human tissue was approved by the ethics committee of the Medical Faculty, University of Yamaguchi Graduate School, and was conducted in accordance with the Declaration of Helsinki, as amended in Somerset West in 1996.…”

Peripheral nerve pericytes modify the blood–nerve barrier function and tight junctional molecules through the secretion of various soluble factors

“…This absence is partly due to the difficulty in isolating a sufficient amount of pericytes from a miniscule amount of endoneurial tissue. Recently, brain and peripheral nerve pericyte cell lines were successfully established from rats (Shimizu et al, 2008). However, brain and peripheral nerve pericytes of human tissue origin are definitely necessary in order to study human BBB and BNB function.…”

Active form of vitamin D directly protects the blood–brain barrier in multiple sclerosis

“…[23][24][25][26][27] TY10 cells were cultured with culture medium containing EBM-2 (Lonza, Walkersville, MD, USA) supplemented with 20% fetal bovine serum (FBS; Sigma, St. Louis, MO, USA), EGM-2 SingleQuot Kit Suppl. & Growth Factors (Lonza), 100 U/mL penicillin (Sigma), 100 lL/mL streptomycin (Sigma) and 25 ng/mL amphotericin B (Invitrogen, San Francisco, CA, USA), in a CO 2 incubator at 33°C before reaching the confluence.…”