0000000000084217

AUTHOR

Ana M. Blanco

showing 3 related works from this author

A FRET-based assay for characterization of alternative splicing events using peptide nucleic acid fluorescence in situ hybridization

2009

We describe a quantitative method for detecting RNA alternative splicing variants that combines in situ hybridization of fluorescently labeled peptide nucleic acid (PNA) probes with confocal microscopy Förster resonance energy transfer (FRET). The use of PNA probes complementary to sequences flanking a given splice junction allows to specifically quantify, within the cell, the RNA isoform generating such splice junction by FRET measure. As a proof of concept we analyzed two alternative splicing events originating from lymphocyte antigen 6 (LY6) complex, locus G5B (LY6G5B) pre-mRNA. These are characterized by the removal of the first intron (Fully Spliced Isoform, FSI) or by retention of suc…

Peptide Nucleic AcidsGene isoformCytoplasmIn situ hybridizationBiologychemistry.chemical_compoundFluorescence Resonance Energy TransferGeneticsmedicineHumansProtein IsoformsspliceRNA MessengerIn Situ Hybridization FluorescenceMicroscopy ConfocalPeptide nucleic acidmedicine.diagnostic_testAlternative splicingIntronPepsin AAlternative SplicingNucleic Acid ProbesFörster resonance energy transferBiochemistrychemistryBiophysicsMethods OnlineCell NucleolusHeLa CellsFluorescence in situ hybridizationNucleic Acids Research
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Intermittent ethanol exposure induces inflammatory brain damage and causes long-term behavioural alterations in adolescent rats

2007

Adolescent brain development seems to be important for the maturation of brain structures and behaviour. Intermittent binge ethanol drinking is common among adolescents, and this type of drinking can induce brain damage. Because we have demonstrated that chronic ethanol treatment induces inflammatory processes in the brain, we investigate whether intermittent ethanol intoxication enhances cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in adolescent rats, and whether these mediators induce brain damage and cause permanent cognitive dysfunctions. Adolescent rats were exposed to ethanol (3.0 g/kg) for two consecutive days at 48-h intervals over 14 days. Levels of COX-2, iN…

medicine.medical_specialtyCerebellumProgrammed cell deathIndomethacinHippocampusNitric Oxide Synthase Type IIInflammationBrain damageMotor ActivityNeuropsychological TestsDiscrimination Learningchemistry.chemical_compoundindomethacinInternal medicineintermittent ethanol intoxicationmedicineAnimalsDrug InteractionsRats WistarAnalysis of VarianceNeocortexEthanolbiologyBehavior AnimalCell DeathEthanolCaspase 3General NeuroscienceAnti-Inflammatory Agents Non-SteroidalBrainRecognition PsychologyRatsNitric oxide synthasemedicine.anatomical_structureEndocrinologychemistryAnimals NewbornneurobehaviourCyclooxygenase 2inflammationAnesthesiabiology.proteinEncephalitisadolescencemedicine.symptomPsychologyPsychomotor Performance
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A practical approach to FRET-based PNA fluorescence in situ hybridization.

2010

Abstract Given the demand for improved methods for detecting and characterizing RNA variants in situ, we developed a quantitative method for detecting RNA alternative splicing variants that combines in situ hybridization of fluorescently labeled peptide nucleic acid (PNA) probes with confocal microscopy Forster resonance energy transfer (FRET). The use of PNA probes complementary to sequences flanking a given splice junction allows to specifically quantify, within the cell, the RNA isoform generating such splice junction as FRET efficiency measure. The FRET-based PNA fluorescence in situ hybridization (FP-FISH) method offers a conceptually new approach for characterizing at the subcellular …

In situPeptide Nucleic AcidsOligonucleotidesIn situ hybridizationBiologyGeneral Biochemistry Genetics and Molecular Biologylaw.inventionchemistry.chemical_compoundConfocal microscopylawmedicineFluorescence Resonance Energy TransferMolecular BiologyIn Situ Hybridization FluorescenceMicroscopy ConfocalPeptide nucleic acidmedicine.diagnostic_testAlternative splicingRNANucleic Acid HybridizationReproducibility of ResultsMolecular biologyAlternative SplicingFörster resonance energy transferchemistrybiological sciencesBiophysicsFluorescence in situ hybridizationMethods (San Diego, Calif.)
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