Differential gene expression in astrocytes from human normal and glaucomatous optic nerve head analyzed by cDNA microarray.
Recent advances in cDNA microarray technology have made it possible to analyze expression of several thousand genes at the same time. Using this technique, gene expression in human astrocytes cultured from glaucomatous and normal optic nerve heads (ONH) was compared. One hundred-fifty genes were differentially expressed more than 5-fold in glaucomatous cell cultures compared with normal. These genes are involved in a number of biological processes, including signal transduction, cell adhesion and proliferation, ECM synthesis, and degradation. Confirmation of differential gene expression was performed by quantitative RT-PCR. Western blots and immunohistochemistry demonstrated gene products in cell cultures or in human ONH tissues. Proliferation, adhesion and migration assays tested physiological responses suggested by differential gene expression. Our study suggests that cultured glaucomatous ONH astrocytes retain in culture many phenotypic characteristics that may be relevant to their role in the pathogenesis of glaucoma and, in general to reactive astrocytes in the CNS. Potential applications of these data include the identification and characterization of signaling pathways involved in astrocyte function, studies of the role of steroid-metabolizing enzymes in the glaucomatous ONH, and further exploration of the role of selected identified genes in experimental animal and in vitro models of glaucoma.
Hernandez
M Rosario
MR
Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, Missouri 63110, USA. hernandezr@vision.wustl.edu
Agapova
Olga A
OA
Yang
Ping
P
Salvador-Silva
Mercedes
M
Ricard
Cynthia S
CS
Aoi
Shun
S
D000368:Aged; D000369:Aged, 80 and over; D001253:Astrocytes / Q000201:enzymology* / Q000473:pathology*; D002465:Cell Movement / Q000235:genetics; D002478:Cells, Cultured; D004798:Enzymes / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D005901:Glaucoma / Q000201:enzymology / Q000235:genetics* / Q000503:physiopathology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006133:Growth Substances / Q000235:genetics; D006801:Humans; D017136:Ion Transport / Q000235:genetics; D008297:Male; D008875:Middle Aged; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D009569:Nitric Oxide / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D009898:Optic Disk / Q000201:enzymology* / Q000473:pathology* / Q000503:physiopathology; D015398:Signal Transduction / Q000235:genetics
D004798:Enzymes; D016326:Extracellular Matrix Proteins; D005904:Glial Fibrillary Acidic Protein; D006133:Growth Substances; D019006:Neural Cell Adhesion Molecules; D009569:Nitric Oxide