title: Gene expression and immunolocalisation of a calcium-activated chloride channel during the stratification of cultivated and developing corneal epithelium.
abstract: The spatial and temporal localisation of a calcium-activated chloride channel (CLCA) and its mRNA was investigated, during the in vivo and in vitro development of stratified epithelia, by fluorescence immunohistochemistry and quantitative polymerase chain reaction in embryonic chicken corneas and the expansion of excised human corneal stem cells on amniotic membrane. Single-layered human epithelial cultures on amniotic membrane and early day embryonic chicken corneas expressed relatively little human CLCA2 or its chicken homologue. However, as the epithelium in both models matured and the number of cell-layers increased, the gene expression level and protein staining intensity increased, primarily within the basal cells of both the cultured and embryonic tissues. These results demonstrate that human CLCA2 protein and mRNA expression are elevated during epithelial stratification, suggesting that this protein plays a role in the growth of multi-layered corneal epithelia during both natural development and tissue cultivation.
authors.affiliation: Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan. connoncj@cf.ac.uk
authors.forename: Che J
authors.initials: CJ
authors.lastname: Connon
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kawasaki
authors.forename: Melody
authors.initials: M
authors.lastname: Liles
authors.forename: Noriko
authors.initials: N
authors.lastname: Koizumi
authors.forename: Kenta
authors.initials: K
authors.lastname: Yamasaki
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakamura
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Quantock
authors.forename: Shigeru
authors.initials: S
authors.lastname: Kinoshita
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology / Q000378:metabolism; D018118:Chloride Channels / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: C120066:CLCA2 protein, human; D018118:Chloride Channels