Dataset Information

A proteomic map of the human aqueous humor

ABSTRACT: The aqueous humor is a colourless, transparent fluid that fills the anterior chamber of the eye. It has an important role to play in maintaining the intraocular pressure of the eye and providing nourishment to avascular structures such as the lens and cornea. It is also essential for providing a clear path for the entry and exit of light from the eye and maintaining the refractive power of the eye. In this study, 250 samples of aqueous humor were collected from patients undergoing cataract surgery. The samples were pooled, divided into aliquots and subjected to ingel digestion, insolution digestion followed by strong cation exchange chromatography and basic reverse phase liquid chromatography fractionation. MS/MS analysis was carried out on a Fourier transform LTQ-Orbitrap velos mass spectrometer and the data were searched with the search algorithms Mascot, SEQUEST and X!Tandem against the NCBI RefSeq human protein database. We have identified 818 proteins of which 447 have been identified for the first time. Proteins such as sorbitol dehydrogenase, filensin and phakinin were some of the novel proteins identified in this study. Classification of proteins based on domain information yielded 304 proteins with signal peptides, 46 proteins with transmembrane domain and 75 proteins with both signal peptide and transmembrane domains. Our study provides a detailed profile of the human aqueous humor. The results from this study of aqueous proteins should help facilitate research into pathological conditions of the eye such as glaucoma, myopia, cataract and uveitis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Akhilesh Pandey

LAB HEAD: Akhilesh Pandey

PROVIDER: PXD000787 | Pride | 2016-01-04

REPOSITORIES: pride

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Dataset's files

Source:

			Action	DRS
	AH_SCX_all_20ppm_Sequest_Mascot_PV.msf	Msf
	AH_SCX_all_20ppm_Sequest_Mascot_PV.pep.xml	Pepxml
	AH_SCX_all_20ppm_Sequest_Mascot_PV.prot.xml	Xml
	AH_bRP_all_20ppm_Sequest_Mascot_PV.msf	Msf
	AH_bRP_all_20ppm_Sequest_Mascot_PV.pep.xml	Pepxml

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Proteomics of human aqueous humor.

Murthy Krishna R KR Rajagopalan Pavithra P Pinto Sneha M SM Advani Jayshree J Murthy Praveen R PR Goel Renu R Subbannayya Yashwanth Y Balakrishnan Lavanya L Dash Mahashweta M Anil Abhijith K AK Manda Srikanth S SS Nirujogi Raja Sekhar RS Kelkar Dhanashree S DS Sathe Gajanan J GJ Dey Gourav G Chatterjee Aditi A Gowda Harsha H Chakravarti Shukti S Shankar Subramanian S Sahasrabuddhe Nandini A NA Nair Bipin B Somani Babu Lal BL Prasad T S Keshava TS Pandey Akhilesh A

Omics : a journal of integrative biology 20150501 5

The aqueous humor is a colorless, transparent fluid that fills the anterior chamber of the eye. It plays an important role in maintaining the intraocular pressure and providing nourishment to the lens and cornea. The constitution of the aqueous humor is controlled by the blood-aqueous barrier. Though this ocular fluid has been extensively studied, its role in ocular physiology is still not completely understood. In this study, aqueous humor samples were collected from 250 patients undergoing cat ...[more]

PMID: 25933257

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Project description: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 Two condition experiment: Human trabecular mesh work cells infected with Adenivirus expressing GFP Vs Adenovirus expressing GFP and constitutively active RhoAV14