title: Gene Expression Profile of Extracellular Matrix and Adhesion Molecules in the Human Normal Corneal Stroma.
abstract: There is limited information on region-specific gene expression in the human corneal stroma. In this study, we aimed to investigate the expression profile of the extracellular matrix and adhesion molecules in the normal corneal stroma using laser capture microdissection (LCM) and molecular techniques.
  
  
  
  Frozen sections of human cornea without ocular disease were used to isolate the central and peripheral corneal stromal keratocytes by LCM. RNA was extracted from LCM-captured tissues and the RT2Profiler PCR Arrays were used to examine the expression profile of extracellular matrix and adhesion molecules in the central and peripheral stroma. Real-time quantitative PCR was used to quantify gene expression. Proteomic and western blotting (WB) analyses were performed to confirm gene expression at protein level. Function association network was generated via the web tools String and Cytoscape.
  
  
  
  The gene expression profiling demonstrated that 35 out of the 84 extracellular matrix and adhesion molecules represented in the array were expressed in stromal keratocytes. Among them, 24 genes were not previously described in the corneal stroma. Two genes were found more abundantly expressed in the central stroma than in the periphery: TGFBI, COL6A2 (p < 0.05). ADAMTS13 was detected only in the central stroma. Proteomics and WB analysis confirmed the expression of 10 genes. Functional analysis revealed that most identified genes were presented in a core cluster that had multiple and strong associations with other genes.
  
  
  
  This study identified genes not previously described in the corneal stroma, revealed regional differences in gene expression between central and peripheral stroma, and also detected some interesting candidate genes that may play important roles in corneal function. These observations serve as the foundation to further investigate the molecular and cellular mechanisms regulating the pathogenesis of regional corneal stromal disorders such as keratoconus.
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.|c Aier Eye Hospital , Changsha , Hunan , China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.
authors.forename: Hu
authors.initials: H
authors.lastname: Huang
authors.affiliation: d Joint Shantou International Eye Center , Shantou University and the Chinese University of Hong Kong , Shantou , China.
authors.forename: Guoying
authors.initials: G
authors.lastname: Sun
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.|e Department of Ophthalmology , College of Medicine, King Saud University , Riyadh , Saudi Arabia.
authors.forename: Saeed
authors.initials: S
authors.lastname: Alwadani
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Semba
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.
authors.forename: Gerard A
authors.initials: GA
authors.lastname: Lutty
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.
authors.forename: Samuel
authors.initials: S
authors.lastname: Yiu
authors.affiliation: a Wilmer Eye Institute , Johns Hopkins University School of Medicine , Baltimore , MD , USA.|b King Khaled Eye Specialist Hospital , Riyadh , Saudi Arabia.
authors.forename: Deepak P
authors.initials: DP
authors.lastname: Edward
mesh_terms: D000328:Adult; D000368:Aged; D015153:Blotting, Western; D015815:Cell Adhesion Molecules / Q000235:genetics*; D060527:Corneal Keratocytes / Q000378:metabolism*; D003319:Corneal Stroma / Q000166:cytology / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling; D064368:Healthy Volunteers; D006801:Humans; D046228:Microarray Analysis; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D040901:Proteomics; D053719:Tandem Mass Spectrometry; D014019:Tissue Donors
chemical_list: D015815:Cell Adhesion Molecules; D016326:Extracellular Matrix Proteins
keywords: Adhesion molecules; corneal stroma; extracellular matrix; gene expression; laser capture microdissection