RRC ID 44206
Author Hogerheyde TA, Stephenson SA, Harkin DG, Bray LJ, Madden PW, Woolf MI, Richardson NA.
Title Evaluation of Eph receptor and ephrin expression within the human cornea and limbus.
Journal Exp Eye Res
Abstract Eph receptor tyrosine kinases and their ligands, the ephrins, regulate the development and maintenance of multiple organs but little is known about their potential role within the cornea. The purpose of this study was to perform a thorough investigation of Eph/ephrin expression within the human cornea including the limbal stem cell niche. Initially, immunohistochemistry was performed on human donor eyes to determine the spatial distribution of Eph receptors and ephrins in the cornea and limbus. Patterns of Eph/ephrin gene expression in (1) immortalised human corneal endothelial (B4G12) or corneal epithelial (HCE-T) cell lines, and (2) primary cultures of epithelial or stromal cells established from the corneal limbus of cadaveric eye tissue were then assessed by reverse transcription (RT) PCR. Limbal epithelial or stromal cells from primary cultures were also assessed for evidence of Eph/ephrin-reactivity by immunofluorescence. Immunoreactivity for ephrinA1 and EphB4 was detected in the corneal endothelium of donor eyes. EphB4 was also consistently detected in the limbal and corneal epithelium and in cells located in the stroma of the peripheral cornea. Expression of multiple Eph/ephrin genes was detected in immortalised corneal epithelial and endothelial cell lines. Evidence of Eph/ephrin gene expression was also demonstrated in primary cultures of human limbal stromal (EphB4, B6; ephrinA5) and epithelial cells (EphA1, A2; ephrinA5, B2) using both RT-PCR and immunofluorescence. The expression of Eph receptors and ephrins within the human cornea and limbus is much wider than previously appreciated and suggests multiple potential roles for these molecules in the maintenance of normal corneal architecture.
Volume 107
Pages 110-20
Published 2013-2-1
DOI 10.1016/j.exer.2012.11.016
PII S0014-4835(12)00343-0
PMID 23247085
MeSH Cells, Cultured Cornea / metabolism* DNA Primers / chemistry Endothelium, Corneal / metabolism Ephrins / genetics* Ephrins / metabolism Epithelium, Corneal / metabolism Fluorescent Antibody Technique, Indirect Gene Expression Regulation / physiology* Humans Immunoenzyme Techniques Limbus Corneae / metabolism* Polymerase Chain Reaction RNA, Messenger / metabolism Receptors, Eph Family / genetics* Receptors, Eph Family / metabolism Reverse Transcriptase Polymerase Chain Reaction Tissue Donors
IF 3.011
Times Cited 7
Human and Animal Cells HCE-T(RCB2280)