Changes of gap and tight junctions during differentiation of human nasal epithelial cells using primary human nasal epithelial cells and primary human nasal fibroblast cells in a noncontact coculture system.
The epithelium of upper respiratory tissues such as nasal mucosa forms a continuous barrier to a wide variety of exogenous antigens. The epithelial barrier function is regulated in large part by the intercellular junctions, referred to as gap and tight junctions. However, changes of gap and tight junctions during differentiation of human nasal epithelial (HNE) cells are still unclear. In the present study, to investigate changes of gap and tight junctions during differentiation of HNE cells in vitro, we used primary human HNE cells cocultured with primary human nasal fibroblast (HNF) cells in a noncontact system. In HNE cells cocultured with HNF cells for 2 weeks, numerous elongated cilia-like structures were observed compared to those without HNF cells. In the coculture, downregulation of Cx26 and upregulation of Cx30.3 and Cx31 were observed together with extensive gap junctional intercellular communication. Furthermore, expression of the tight junction proteins claudin-1, claudin-4, occludin and ZO-2 was increased. These results suggest that switching in expression of connexins and induction of tight junction proteins may be closely associated with differentiation of HNE cells in vitro and that differentiation of HNE cells requires unknown soluble factors secreted from HNF cells.
Koizumi
Jun-ichi
J
Department of Otolaryngology, Sapporo Medical University School of Medicine, S1. W17, Sapporo, Japan.
Kojima
Takashi
T
Kamekura
Ryuta
R
Kurose
Makoto
M
Harimaya
Atsushi
A
Murata
Masaki
M
Osanai
Makoto
M
Chiba
Hideki
H
Himi
Tetsuo
T
Sawada
Norimasa
N
D015153:Blotting, Western; D002450:Cell Communication / Q000528:radiation effects; D002454:Cell Differentiation; D002478:Cells, Cultured; D062445:Claudin-1; D062506:Claudin-4; D018920:Coculture Techniques; D000072259:Connexin 26; D017630:Connexins / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D017629:Gap Junctions / Q000502:physiology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007365:Intercellular Junctions; D008565:Membrane Proteins / Q000378:metabolism; D009297:Nasal Mucosa / Q000166:cytology* / Q000378:metabolism; D062793:Occludin; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions / Q000502:physiology*; D062827:Zonula Occludens-2 Protein
C568108:CLDN1 protein, human; C568157:CLDN4 protein, human; D062445:Claudin-1; D062506:Claudin-4; D017630:Connexins; C000607042:GJB2 protein, human; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D012333:RNA, Messenger; C569046:TJP2 protein, human; D062827:Zonula Occludens-2 Protein; D000072259:Connexin 26; C068044:GJB3 protein, human; C076275:connexin 30.3