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Series GSE9288 Query DataSets for GSE9288
Status Public on Sep 25, 2008
Title Constitutively active RhoA (RhoAV14) expression and regulation of acqeous humor outflow dynamics
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Impaired drainage of aqueous humor through the trabecular meshwork (TM) culminating in increased intraocular pressure is a major risk factor for glaucoma, a leading cause of blindness worldwide. Regulation of aqueous humor drainage through the TM, however, is poorly understood. The role of RhoA GTPase-mediated contractile activity, cell adhesive interactions, and gene expression in regulation of aqueous humor outflow was investigated using adenoviral vector-driven expression of constitutively active RhoA (RhoAV14). Organ cultured anterior segments from porcine eyes expressing RhoAV14 exhibited significant reduction of aqueous humor outflow. Cultured TM cells expressing RhoAV14 revealed strong contractile cell morphology, increased actin stress fibers and focal adhesions, along with increased levels of phosphorylated myosin II, and collagen IV, fibronectin and laminin. cDNA microarray analysis of RNA extracted from RhoAV14 expressing human TM cells revealed a significant increase in the expression of genes encoding extracellular matrix (ECM) proteins, cytokines, integrins, cytoskeletal proteins and signaling proteins. Conversely, various ECM proteins stimulated robust increases in phosphorylation of myosin II, paxillin and focal adhesion kinase, and activated Rho GTPase and actin stress fiber formation in TM cells, indicating a potential regulatory feedback interaction between ECM-induced mechanical strain and Rho GTPase-induced isometric tension in TM cells. Collectively, these data demonstrate that sustained activation of Rho GTPase signaling in the aqueous humor outflow pathway increases resistance to aqueous humor outflow through the trabecular pathway by influencing the contractile force, cell adhesive interactions, and the expression of ECM proteins and cytokines in TM cells.
Keywords: Gene Expression
Overall design Two condition experiment: Human trabecular mesh work cells infected with Adenivirus expressing GFP Vs Adenovirus expressing GFP and constitutively active RhoAV14
Contributor(s) Rao VP
Citation(s) 18799648
Submission date Oct 10, 2007
Last update date Jan 18, 2013
Contact name Vasantha P Rao
Phone 919-681-5883
Fax 919-684-8983
Organization name Duke University Medical Center
Department Ophthalmology
Lab Glaucoma
Street address 2351 Erwin Rd
City Durham
State/province NC
ZIP/Postal code 27710
Country USA
Platforms (1)
GPL5920 Duke University Human Operon v.3.0 spotted Array
Samples (2)
GSM236674 Human primary TM cells expressing Adenivirus GFP
GSM236675 Human primary TM cells expressing Adenivirus GFP with Constitutively active RhoA
BioProject PRJNA102925

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE9288_RAW.tar 6.9 Mb (http)(custom) TAR (of GPR)
GSE9288_hyb_protocol.pdf 32.2 Kb (ftp)(http) PDF
GSE9288_labeling_protocol.pdf 25.3 Kb (ftp)(http) PDF
Processed data included within Sample table

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