title: Identification and dynamic regulation of tight junction protein expression in human neural stem cells.
abstract: Recent reports indicate that neural stem cells (NSCs) exist in a cluster-like formation in close proximity to cerebral microvessels. Similar appearing clusters can be seen ex vivo in NSC cultures termed neurospheres. It is known that this neurosphere configuration is important for preserving stemness and a proliferative state. How NSCs form neurospheres or neuroclusters remains largely undetermined. In this study, we show that primary human NSCs express the tight junction proteins (TJPs): zonula occludens-1 (ZO-1), occludin, claudin-1, -3, -5, and -12. The relative mRNA expression was measured by quantitative polymerase chain reaction, and protein expression was confirmed by flow cytometry and immunofluorescence microscopy. Our results show that downregulation of TJPs occurs as neuronal differentiation is induced, suggesting that control of TJPs may be tied to the neuronal differentiation program. Importantly, upon specific knockdown of the accessory TJP, ZO-1, undifferentiated NSCs showed decreased levels of key stem cell markers. Taken together, our results indicate that TJPs possibly aid in maintaining the intercellular configuration of NSCs and that reduction in TJP expression consequently affects the stemness status.
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Andrea K
authors.initials: AK
authors.lastname: Watters
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Slava
authors.initials: S
authors.lastname: Rom
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Jeremy D
authors.initials: JD
authors.lastname: Hill
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Marie K
authors.initials: MK
authors.lastname: Dematatis
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Steven F
authors.initials: SF
authors.lastname: Merkel
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Allison M
authors.initials: AM
authors.lastname: Andrews
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Cena
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.|2Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Raghava
authors.initials: R
authors.lastname: Potula
authors.affiliation: 3Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Andrew
authors.initials: A
authors.lastname: Skuba
authors.affiliation: 3Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Young-Jin
authors.initials: YJ
authors.lastname: Son
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.|2Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Yuri
authors.initials: Y
authors.lastname: Persidsky
authors.affiliation: 1Department of Pathology and Laboratory Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania.|2Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, Pennsylvania.|3Shriners Hospitals Pediatric Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania.
authors.forename: Servio H
authors.initials: SH
authors.lastname: Ramirez
mesh_terms: D002454:Cell Differentiation / Q000235:genetics*; D062445:Claudin-1 / Q000096:biosynthesis; D062465:Claudin-3 / Q000096:biosynthesis; D062507:Claudin-5 / Q000096:biosynthesis; D057167:Claudins / Q000096:biosynthesis; D005434:Flow Cytometry; D018507:Gene Expression Regulation, Developmental; D000066449:Human Embryonic Stem Cells / Q000378:metabolism; D006801:Humans; D058953:Neural Stem Cells / Q000378:metabolism*; D062793:Occludin / Q000096:biosynthesis; D062725:Tight Junction Proteins / Q000096:biosynthesis*; D019108:Tight Junctions / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000096:biosynthesis / Q000235:genetics*
chemical_list: C522352:CLDN12 protein, human; C107466:CLDN5 protein, human; D062445:Claudin-1; D062465:Claudin-3; D062507:Claudin-5; D057167:Claudins; C568985:OCLN protein, human; D062793:Occludin; C569042:TJP1 protein, human; D062725:Tight Junction Proteins; D062826:Zonula Occludens-1 Protein