Unique distribution of thrombospondin-1 in human ocular surface epithelium.
OBJECTIVE
The study was conducted to elucidate the detailed expression pattern of angiogenesis-related factors in human ocular surface epithelium. The focus was factors with significantly higher gene expression in corneal epithelium (CE) than in conjunctival epithelium (CJE).


METHODS
The relative gene expression of 36 angiogenesis-related factors was compared in human CE and CJE, by using the introduced amplified fragment-length polymorphism (iAFLP) METHOD: Also examined were the expression patterns in the CE, limbal epithelium (LE), and CJE of factors with significantly higher expression in the CE, by using real-time PCR, in situ hybridization, immunohistochemistry, and immunoelectron microscopy.


RESULTS
Only thrombospondin (TSP)-1 exhibited significantly higher expression in the CE. In situ hybridization and real-time PCR showed TSP-1 transcripts in the basal cells of the CE and LE. Compared with the CJE, they were significantly upregulated at those sites. Immunohistochemistry revealed that TSP-1 was strongly expressed in the basal region of the CE. Its expression was faint in LE and absent in CJE. Immunoelectron microscopy revealed that the CE and LE demonstrated TSP-1 labeling just below the epithelium, in the basal region of basal cells, and occasionally in the basal cell membrane. There was little or no labeling in the CJE.


CONCLUSIONS
In the human ocular surface epithelium, basal cells of the CE and LE, but not of the CJE, synthesize TSP-1. High levels of TSP-1 are present only just below the CE. Its unique distribution may be related to corneal avascularity and integrity.
Sekiyama
Eiichi
E
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kyoto, Japan. esekiyam@ophth.kpu-m.ac.jp
Nakamura
Takahiro
T
Cooper
Leanne J
LJ
Kawasaki
Satoshi
S
Hamuro
Junji
J
Fullwood
Nigel J
NJ
Kinoshita
Shigeru
S
D042501:Angiogenic Proteins / Q000235:genetics / Q000378:metabolism; D003228:Conjunctiva / Q000378:metabolism* / Q000648:ultrastructure; D017931:DNA Primers / Q000737:chemistry; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D019573:Epithelium, Corneal / Q000378:metabolism* / Q000648:ultrastructure; D005784:Gene Amplification; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D016253:Microscopy, Immunoelectron; D008875:Middle Aged; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019700:Thrombospondin 1 / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D015854:Up-Regulation
D042501:Angiogenic Proteins; D017931:DNA Primers; D019700:Thrombospondin 1