2018
The Zinc-Metallothionein Redox System Reduces Oxidative Stress in Retinal Pigment Epithelial Cells
Rodríguez-Menéndez S, García M, Fernández B, Álvarez L, Fernández-Vega-Cueto A, Coca-Prados M, Pereiro R, González-Iglesias H. The Zinc-Metallothionein Redox System Reduces Oxidative Stress in Retinal Pigment Epithelial Cells. Nutrients 2018, 10: 1874. PMID: 30513827, PMCID: PMC6315569, DOI: 10.3390/nu10121874.Peer-Reviewed Original ResearchConceptsRetinal pigment epitheliumOxidative stressRetinal pigment epithelial cellsOxidative damagePre-treated cellsInduces oxidative stressPigment epithelial cellsNon-treated cellsReactive oxygen intermediatesΜM of zincPigment epitheliumRPE cellsZn-MTProtective mechanismEpithelial cellsFree radical generatorMT levelsAAPH treatmentOxygen intermediates
2014
Characterization of Vitamin D Production by Human Ocular Barrier CellsVitamin D Production by Human Ocular Barrier Cells
Alsalem JA, Patel D, Susarla R, Coca-Prados M, Bland R, Walker EA, Rauz S, Wallace GR. Characterization of Vitamin D Production by Human Ocular Barrier CellsVitamin D Production by Human Ocular Barrier Cells. Investigative Ophthalmology & Visual Science 2014, 55: 2140-2147. PMID: 24576880, DOI: 10.1167/iovs.13-13019.Peer-Reviewed Original ResearchConceptsHuman scleral fibroblastsVitamin D3Vitamin D receptorBarrier epithelial cellsPrimary human scleral fibroblastsEnzyme immunoassayD productionUltraviolet B sunlightEpithelial cellsVitamin D productionRetinal pigment epithelial cell lineBarrier cellsExtrarenal synthesisHydroxyvitamin D3Epithelial cell lineImmune regulationCiliary bodyD receptorExpression of componentsSkin precursorsODM-2Scleral fibroblastsBarrier sitesCorneal endothelialBarrier function
2013
Bicarbonate-Dependent Secretion and Proteolytic Processing of Recombinant Myocilin
Aroca-Aguilar JD, Martínez-Redondo F, Martín-Gil A, Pintor J, Coca-Prados M, Escribano J. Bicarbonate-Dependent Secretion and Proteolytic Processing of Recombinant Myocilin. PLOS ONE 2013, 8: e54385. PMID: 23342144, PMCID: PMC3547000, DOI: 10.1371/journal.pone.0054385.Peer-Reviewed Original ResearchConceptsIrreversible visual lossElevated intraocular pressureIntracellular accumulationC-terminal fragmentVisual lossOptic neuropathyIntraocular pressureIntracellular proteolytic processingProteolytic processingOcular tissuesRecombinant myocilinCalpain IICellular modelMyocilinSecretionPossible factorsExtracellular glycoproteinNeuropathyGlaucoma
2011
Interaction of Recombinant Myocilin with the Matricellular Protein SPARC: Functional Implications
Aroca-Aguilar JD, Sánchez-Sánchez F, Ghosh S, Fernández-Navarro A, Coca-Prados M, Escribano J. Interaction of Recombinant Myocilin with the Matricellular Protein SPARC: Functional Implications. Investigative Ophthalmology & Visual Science 2011, 52: 179-189. PMID: 20926826, PMCID: PMC3053273, DOI: 10.1167/iovs.09-4866.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceBlotting, WesternCalcium-Binding ProteinsCell LineChromatography, High Pressure LiquidCytoskeletal ProteinsExtracellular Matrix ProteinsEye ProteinsGene Expression ProfilingGlycoproteinsHumansKidneyMicroscopy, FluorescenceMolecular Sequence DataOsteonectinPolymerase Chain ReactionProtein BindingRecombinant Fusion ProteinsRecombinant ProteinsTwo-Hybrid System TechniquesConceptsHEK 293T cellsSolid-phase binding assaysInteraction of myocilinRecombinant proteinsRecombinant myocilinMatricellular proteinEC domainTerminal olfactomedin domainTwo-hybrid analysisTwo-hybrid systemProtein-protein interactionsFull-length myocilinC-terminal domainN-terminal domainC-terminal regionCalcium binding domainsBinding assaysOlfactomedin domainC-terminal fragmentBinding domainsLinker regionUnknown functionExtracellular glycoproteinIntracellular interactionsSPARC family
2009
Functional Role of Proteolytic Processing of Recombinant Myocilin in Self-Aggregation
Aroca-Aguilar JD, Martínez-Redondo F, Sánchez-Sánchez F, Coca-Prados M, Escribano J. Functional Role of Proteolytic Processing of Recombinant Myocilin in Self-Aggregation. Investigative Ophthalmology & Visual Science 2009, 51: 72-78. PMID: 19696176, PMCID: PMC2869055, DOI: 10.1167/iovs.09-4118.Peer-Reviewed Original Research
2008
Heterozygous expression of myocilin glaucoma mutants increases secretion of the mutant forms and reduces extracellular processed myocilin.
Aroca-Aguilar JD, Sánchez-Sánchez F, Martínez-Redondo F, Coca-Prados M, Escribano J. Heterozygous expression of myocilin glaucoma mutants increases secretion of the mutant forms and reduces extracellular processed myocilin. Molecular Vision 2008, 14: 2097-108. PMID: 19023451, PMCID: PMC2585175.Peer-Reviewed Original ResearchConceptsWild-type myocilinWild-type proteinMyocilin mutantsMutant myocilinMutant formsProteolytic processingMissense mutant formsHEK 293T cellsMyocilin geneMutant proteinsSecretory pathwayUnidentified functionExtracellular proteinsMutantsEndoproteolytic processingRecombinant mutantsMyocilin secretionCellular fractionsHeterozygous expressionMyocilinProteinUnknown mechanismExtracellular amountSDS-PAGEHeterozygous stateExpression and purification of functional recombinant human pigment epithelium-derived factor (PEDF) secreted by the yeast Pichia pastoris
Sánchez-Sánchez F, Aroca-Aguilar JD, Segura I, Ramírez-Castillejo C, Riese HH, Coca-Prados M, Escribano J. Expression and purification of functional recombinant human pigment epithelium-derived factor (PEDF) secreted by the yeast Pichia pastoris. Journal Of Biotechnology 2008, 134: 193-201. PMID: 18282627, DOI: 10.1016/j.jbiotec.2008.01.005.Peer-Reviewed Original ResearchConceptsPigment epithelium-derived factorEpithelium-derived factorRecombinant pigment epithelium-derived factorRecombinant human pigment epithelium-derived factorHuman pigment epithelium-derived factorCerebellar granule cell survivalPotential therapeutic agentGranule cell survivalFull-length PEDFStem cell self-renewal propertiesCell-based therapiesTherapeutic roleOcular diseasesTherapeutic agentsSelf-renewal propertiesEndothelial cell migrationLiquid chromatographyCell linesRPEDFCell survivalHigh-performance liquid chromatographyCell migrationYeast Pichia pastorisLow pressure liquid chromatographyPichia pastoris
2007
Characterization of the Intracellular Proteolytic Cleavage of Myocilin and Identification of Calpain II as a Myocilin-processing Protease*
Sánchez-Sánchez F, Martínez-Redondo F, Aroca-Aguilar JD, Coca-Prados M, Escribano J. Characterization of the Intracellular Proteolytic Cleavage of Myocilin and Identification of Calpain II as a Myocilin-processing Protease*. Journal Of Biological Chemistry 2007, 282: 27810-27824. PMID: 17650508, DOI: 10.1074/jbc.m609608200.Peer-Reviewed Original ResearchConceptsExtracellular calciumCalpain IICalcium-activated proteaseIntraocular pressureT cellsIntracellular proteolytic cleavageCalpain inhibitorsCalcium uptakeProteolytic cleavageCalpain inhibitor IVOlfactomedin-like domainCalpain IInhibitor IVMyocilinEndoplasmic reticulumIntracellular processingLumenRNA interference knockdownCalciumProteolytic processingCellsCulture mediumGlaucomaSubcellular fractionationEndoproteolytic processingRole of MYOC and OPTN sequence variations in Spanish patients with primary open-angle glaucoma.
López-Martínez F, López-Garrido M, Sánchez-Sánchez F, Campos-Mollo E, Coca-Prados M, Escribano J. Role of MYOC and OPTN sequence variations in Spanish patients with primary open-angle glaucoma. Molecular Vision 2007, 13: 862-72. PMID: 17615537, PMCID: PMC2770203.Peer-Reviewed Original ResearchMeSH KeywordsAge of OnsetAmino Acid SequenceCell Cycle ProteinsCell LineCytoskeletal ProteinsExonsEye ProteinsFemaleGene FrequencyGenome, HumanGlaucoma, Open-AngleGlycoproteinsHaplotypesHumansLinkage DisequilibriumMaleMembrane Transport ProteinsMiddle AgedMolecular Sequence DataMutationOcular HypertensionOpen Reading FramesPhenotypePolymorphism, Single-Stranded ConformationalPromoter Regions, GeneticSpainTranscription Factor TFIIIAWhite PeopleConceptsPrimary open-angle glaucomaOcular hypertensionOpen-angle glaucomaPOAG patientsSpanish patientsSequence variationExons of myocilinHeterozygous disease-causing mutationsPathogenic mutationsSingle nucleotide polymorphismsPromoter regionRole of myocilinDNA sequence variationPolymorphic GT microsatelliteCommon single nucleotide polymorphismsPCR-DNA sequencingT cellsComplete coding regionPatientsUnrelated patientsMYOC geneOPTN geneGlaucomaDisease-causing mutationsGT microsatellite
2005
Interaction of myocilin with the C-terminal region of hevin
Li Y, Aroca-Aguilar JD, Ghosh S, Sánchez-Sánchez F, Escribano J, Coca-Prados M. Interaction of myocilin with the C-terminal region of hevin. Biochemical And Biophysical Research Communications 2005, 339: 797-804. PMID: 16316624, DOI: 10.1016/j.bbrc.2005.11.082.Peer-Reviewed Original ResearchMyocilin Mutations Causing Glaucoma Inhibit the Intracellular Endoproteolytic Cleavage of Myocilin between Amino Acids Arg226 and Ile227 *
Aroca-Aguilar JD, Sánchez-Sánchez F, Ghosh S, Coca-Prados M, Escribano J. Myocilin Mutations Causing Glaucoma Inhibit the Intracellular Endoproteolytic Cleavage of Myocilin between Amino Acids Arg226 and Ile227 *. Journal Of Biological Chemistry 2005, 280: 21043-21051. PMID: 15795224, DOI: 10.1074/jbc.m501340200.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid SequenceAnimalsArginineBlotting, WesternBrefeldin ACattleCell LineComputational BiologyCOS CellsCulture MediaCytoskeletal ProteinsExtracellular Matrix ProteinsEye ProteinsGlaucomaGlycoproteinsGreen Fluorescent ProteinsHumansIsoleucineLeucineMicroscopy, FluorescenceMolecular Sequence DataMutationPeptidesPhenotypeProtein Structure, TertiarySpectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationTransfectionConceptsMyocilin mutationPathogenesis of glaucomaCause of blindnessSevere glaucoma phenotypeHuman aqueous humorAqueous humorOcular tissuesGlaucoma phenotypeNonocular tissuesWestern immunoblot analysisGlaucomaPathogenic mutationsOlfactomedin-like domainEndoproteolytic processingWild-type myocilinCell linesImmunoblot analysisMyocilinEndoproteolytic cleavageLeucine zipper-like domainMutant myocilinNormal roleHuman organsInhibitionTissue
2004
Hyposmotic activation of ICl,swell in rabbit nonpigmented ciliary epithelial cells involves increased ClC-3 trafficking to the plasma membrane
Vessey JP, Shi C, Jollimore CA, Stevens KT, Coca-Prados M, Barnes S, Kelly ME. Hyposmotic activation of ICl,swell in rabbit nonpigmented ciliary epithelial cells involves increased ClC-3 trafficking to the plasma membrane. Biochemistry And Cell Biology 2004, 82: 708-718. PMID: 15674438, DOI: 10.1139/o04-107.Peer-Reviewed Original ResearchConceptsPlasma membraneHyposmotic stimulationFluorescent membrane dye FM1-43Membrane dye FM1-43Trafficking of channelsClC-3 channelsCiliary epithelial cellsPhosphoinositide-3 kinase inhibitor wortmanninRate of exocytosisClC-3 Cl(-) channelsEpithelial cellsVesicular traffickingDye FM1-43Membrane dynamicsNonpigmented ciliary epithelial cellsInhibitor wortmanninClC-3 antisenseImmunofluorescence microscopyTraffickingClC-3Regulatory volume decreaseFM1-43Cl- channelsMembraneCell swelling
2003
Production, purification, and functional analysis of recombinant human and mouse 17β-hydroxysteroid dehydrogenase type 7
Törn S, Nokelainen P, Kurkela R, Pulkka A, Menjivar M, Ghosh S, Coca-Prados M, Peltoketo H, Isomaa V, Vihko P. Production, purification, and functional analysis of recombinant human and mouse 17β-hydroxysteroid dehydrogenase type 7. Biochemical And Biophysical Research Communications 2003, 305: 37-45. PMID: 12732193, DOI: 10.1016/s0006-291x(03)00694-6.Peer-Reviewed Original ResearchConceptsPeripheral tissuesType 7Primary open-angle glaucomaEarly-onset prostate cancerOpen-angle glaucomaSex steroid hormonesIntracrine regulatorAdrenal glandProstate cancerEstrogenic milieuPotent androgenInactive metabolitesEstrogenic metabolitesSteroid metabolismPotent estrogenSteroid hormonesCancer 1DihydrotestosteroneRecombinant humanTissue distributionHormoneTissueHuman tissuesPresent studyMetabolic roleSerum- and Glucocorticoid-Regulated Kinase Isoform-1 and Epithelial Sodium Channel Subunits in Human Ocular Ciliary Epithelium
Rauz S, Walker EA, Hughes SV, Coca-Prados M, Hewison M, Murray PI, Stewart PM. Serum- and Glucocorticoid-Regulated Kinase Isoform-1 and Epithelial Sodium Channel Subunits in Human Ocular Ciliary Epithelium. Investigative Ophthalmology & Visual Science 2003, 44: 1643-1651. PMID: 12657604, DOI: 10.1167/iovs.02-0514.Peer-Reviewed Original ResearchMeSH KeywordsAldosteroneBlotting, NorthernCell LineCiliary BodyDexamethasoneDose-Response Relationship, DrugDrug CombinationsEpithelial Sodium ChannelsGene Expression RegulationGlucocorticoidsHormone AntagonistsHumansImmediate-Early ProteinsIn Situ HybridizationIsoenzymesMifepristoneMineralocorticoid Receptor AntagonistsNuclear ProteinsPigment Epithelium of EyeProtein Serine-Threonine KinasesReceptors, GlucocorticoidReverse Transcriptase Polymerase Chain ReactionRNA, MessengerSodium ChannelsSpironolactoneConceptsExpression of SGK1Human ocular ciliary epitheliumODM-2 cellsOcular ciliary epitheliumSodium transportCiliary epitheliumENaC subunitsNPE cell linesEpithelial sodium channel (ENaC) subunitsAqueous humor productionSodium channel subunitsExpression of ENaCRadioligand-binding assaysDose-dependent inductionIsoform 1Epithelial sodium channelCorticosteroid regulationHuman ocularAldosteroneCiliary epithelial bilayerRU26752DexamethasoneRT-PCR analysisSodium channelsChannel subunits
2002
Differential regulation of gene expression of neurotensin and prohormone convertases PC1 and PC2 in the bovine ocular ciliary epithelium: possible implications on neurotensin processing
Ortego J, Wollmann G, Coca-Prados M. Differential regulation of gene expression of neurotensin and prohormone convertases PC1 and PC2 in the bovine ocular ciliary epithelium: possible implications on neurotensin processing. Neuroscience Letters 2002, 333: 49-53. PMID: 12401558, DOI: 10.1016/s0304-3940(02)00028-9.Peer-Reviewed Original Research
2001
Molecular Evidence That Human Ocular Ciliary Epithelium Expresses Components Involved in Phototransduction
Bertazolli-Filho R, Ghosh S, Huang W, Wollmann G, Coca-Prados M. Molecular Evidence That Human Ocular Ciliary Epithelium Expresses Components Involved in Phototransduction. Biochemical And Biophysical Research Communications 2001, 284: 317-325. PMID: 11394879, DOI: 10.1006/bbrc.2001.4970.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAntibodies, MonoclonalArrestinBlotting, WesternCalcium-Binding ProteinsCattleCell LineCiliary BodyCyclic Nucleotide-Gated Cation ChannelsEpithelial CellsEye ProteinsFluorescent Antibody Technique, IndirectG-Protein-Coupled Receptor Kinase 1HippocalcinHumansImmunoblottingImmunohistochemistryIon ChannelsLight Signal TransductionLipoproteinsNerve Tissue ProteinsPhosphoric Diester HydrolasesProtein KinasesProtein SubunitsRecoverinRetinaReverse Transcriptase Polymerase Chain ReactionRhodopsinRNA, MessengerSequence Analysis, DNATransducinConceptsHuman ocular ciliary epitheliumCiliary epitheliumOcular ciliary epitheliumNPE cellsCiliary epithelial cell linePars plicata regionIntact ciliary epitheliumWestern blot analysisCultured NPE cellsEpithelial cell lineExpression of rhodopsinIndirect immunofluorescenceVisual arrestinHuman retinaMonoclonal antibodiesHuman ciliary epitheliumThree- to fourfoldEpitheliumRT-PCRPhotoreceptor cellsRhodopsin mRNABlot analysisCell linesRetinaRhodopsin kinase
2000
Tamoxifen and ATP synergistically activate Cl− release by cultured bovine pigmented ciliary epithelial cells
Mitchell C, Peterson‐Yantorno K, Coca‐Prados M, Civan M. Tamoxifen and ATP synergistically activate Cl− release by cultured bovine pigmented ciliary epithelial cells. The Journal Of Physiology 2000, 525: 183-193. PMID: 10811736, PMCID: PMC2269939, DOI: 10.1111/j.1469-7793.2000.00183.x.Peer-Reviewed Original Research
1999
Hyposmotically activated chloride channels in cultured rabbit non‐pigmented ciliary epithelial cells
Shi C, Ryan J, French A, Coca‐Prados M, Kelly M. Hyposmotically activated chloride channels in cultured rabbit non‐pigmented ciliary epithelial cells. The Journal Of Physiology 1999, 521: 57-67. PMID: 10562334, PMCID: PMC2269649, DOI: 10.1111/j.1469-7793.1999.00057.x.Peer-Reviewed Original ResearchMeSH Keywords1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic AcidAnimalsCalciumCell LineChloride ChannelsCiliary BodyEnzyme InhibitorsEpithelial CellsNiflumic AcidOsmotic PressurePatch-Clamp TechniquesPhosphorylationProtein Kinase InhibitorsRabbitsSecond Messenger SystemsStilbenesConceptsNon-pigmented ciliary epithelial cellsWhole-cell currentsCiliary epithelial cellsWhole-cell patch-clamp recording techniquePatch-clamp recording techniquesEpithelial cellsTime courseSlower time courseDisulfonic acidSmall conductanceWhole-cell conductanceCl- current amplitudeExposure of cellsVoltage-sensitive blockCl- currentVoltage-independent blockNiflumic acidRecording techniquesDepolarized potentialsCurrent activationAmplitude of currentsOutward rectificationCell swellingNPCE cellsHyposmotic shockFunctional Expression of Components of the Natriuretic Peptide System in Human Ocular Nonpigmented Ciliary Epithelial Cells
Ortego J, Coca-Prados M. Functional Expression of Components of the Natriuretic Peptide System in Human Ocular Nonpigmented Ciliary Epithelial Cells. Biochemical And Biophysical Research Communications 1999, 258: 21-28. PMID: 10222228, DOI: 10.1006/bbrc.1999.0573.Peer-Reviewed Original ResearchConceptsNatriuretic peptide systemNPR-B receptorNPR-C receptorCiliary epithelial cellsReceptor mRNACiliary epitheliumEpithelial cellsNPE cellsCGMP responseAutocrine/paracrine modulatorPeptide systemHuman ocular ciliary epitheliumNonpigmented ciliary epithelial cellsOcular ciliary epitheliumCultured NPE cellsLong-term downregulationC-ANP4Paracrine modulatorEye pressureBNPActivation of PKCBasal levelsInhibitory effectRT-PCRReceptors
1998
Molecular Identification and Coexpression of Galanin and GalR‐1 Galanin Receptor in the Human Ocular Ciliary Epithelium: Differential Modulation of Their Expression by the Activation of α2‐ and β2‐Adrenergic Receptors in Cultured Ciliary Epithelial Cells
Ortego J, Coca‐Prados M. Molecular Identification and Coexpression of Galanin and GalR‐1 Galanin Receptor in the Human Ocular Ciliary Epithelium: Differential Modulation of Their Expression by the Activation of α2‐ and β2‐Adrenergic Receptors in Cultured Ciliary Epithelial Cells. Journal Of Neurochemistry 1998, 71: 2260-2270. PMID: 9832123, DOI: 10.1046/j.1471-4159.1998.71062260.x.Peer-Reviewed Original ResearchMeSH KeywordsAdrenergic alpha-AgonistsAqueous HumorCell LineCiliary BodyClonidineColforsinCulture Media, Serum-FreeEpithelial CellsFluorescent Antibody TechniqueGalaninHumansNorepinephrineProtein PrecursorsReceptors, Adrenergic, alphaReceptors, Adrenergic, betaReceptors, GalaninReceptors, NeuropeptideRNA, MessengerTetradecanoylphorbol AcetateConceptsGalanin mRNA expression