2018
Aromatase and estrogen receptor immunoreactivity in the coronary arteries of monkeys and human subjects
Diano S, Horvath TL, Mor G, Register T, Adams M, Harada N, Naftolin F. Aromatase and estrogen receptor immunoreactivity in the coronary arteries of monkeys and human subjects. Menopause The Journal Of The North American Menopause Society 2018, 25: 1201-1207. PMID: 30358714, DOI: 10.1097/gme.0000000000001219.Peer-Reviewed Original ResearchConceptsCoronary arteryEstrogen receptor immunoreactivityAmount of atherosclerosisHuman placental estrogen synthetaseHuman subjectsSmooth muscle cellsPrecursor androgensReceptor immunoreactivityCardioprotective effectsCoronary circulationEstrogen formationEstrogen receptorArteryCardiovascular systemER αMuscle cellsEndothelial cellsEstrogenEstrogen synthetaseMonkeysFirst evidenceSubjectsCellsAtherosclerosisLocal regulation
2017
Endothelial HIF-1α Enables Hypothalamic Glucose Uptake to Drive POMC Neurons
Varela L, Suyama S, Huang Y, Shanabrough M, Tschöp M, Gao XB, Giordano FJ, Horvath TL. Endothelial HIF-1α Enables Hypothalamic Glucose Uptake to Drive POMC Neurons. Diabetes 2017, 66: db161106. PMID: 28292966, PMCID: PMC5440016, DOI: 10.2337/db16-1106.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBehavior, AnimalBlotting, WesternEndotheliumEnergy MetabolismFood DeprivationGene Knockdown TechniquesGlucoseHyperphagiaHypothalamusHypoxia-Inducible Factor 1, alpha SubunitImmunohistochemistryMiceMicroscopy, ElectronMitochondriaNeuronsPatch-Clamp TechniquesPro-OpiomelanocortinReal-Time Polymerase Chain ReactionConceptsPOMC neuronsGlucose uptakePOMC neuronal activityHypothalamic proopiomelanocortin (POMC) neuronsHypoxia-inducible factor-1αProopiomelanocortin neuronsVascular impairmentGlucose administrationMetabolic disordersNeuronal activityMetabolic environmentFactor-1αImpaired functioningEndothelial cellsNeuronsFood deprivationVivoCentral controlHypothalamusMiceAdministrationUptakeImpairment
2016
Molecular interrogation of hypothalamic organization reveals distinct dopamine neuronal subtypes
Romanov RA, Zeisel A, Bakker J, Girach F, Hellysaz A, Tomer R, Alpár A, Mulder J, Clotman F, Keimpema E, Hsueh B, Crow AK, Martens H, Schwindling C, Calvigioni D, Bains JS, Máté Z, Szabó G, Yanagawa Y, Zhang MD, Rendeiro A, Farlik M, Uhlén M, Wulff P, Bock C, Broberger C, Deisseroth K, Hökfelt T, Linnarsson S, Horvath TL, Harkany T. Molecular interrogation of hypothalamic organization reveals distinct dopamine neuronal subtypes. Nature Neuroscience 2016, 20: 176-188. PMID: 27991900, PMCID: PMC7615022, DOI: 10.1038/nn.4462.Peer-Reviewed Original Research
2014
Leptin signaling in astrocytes regulates hypothalamic neuronal circuits and feeding
Kim JG, Suyama S, Koch M, Jin S, Argente-Arizon P, Argente J, Liu ZW, Zimmer MR, Jeong JK, Szigeti-Buck K, Gao Y, Garcia-Caceres C, Yi CX, Salmaso N, Vaccarino FM, Chowen J, Diano S, Dietrich MO, Tschöp MH, Horvath TL. Leptin signaling in astrocytes regulates hypothalamic neuronal circuits and feeding. Nature Neuroscience 2014, 17: 908-910. PMID: 24880214, PMCID: PMC4113214, DOI: 10.1038/nn.3725.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAstrocytesCell CountEatingExcitatory Postsynaptic PotentialsGlial Fibrillary Acidic ProteinHypothalamusImmunohistochemistryIn Situ HybridizationLeptinMaleMelanocortinsMiceMice, KnockoutMicroscopy, ElectronNerve NetNeuronsPrimary Cell CulturePro-OpiomelanocortinPulmonary Gas ExchangeReal-Time Polymerase Chain ReactionRNA, MessengerSignal Transduction
2013
Antibodies to cannabinoid type 1 receptor co‐react with stomatin‐like protein 2 in mouse brain mitochondria
Morozov YM, Dominguez MH, Varela L, Shanabrough M, Koch M, Horvath TL, Rakic P. Antibodies to cannabinoid type 1 receptor co‐react with stomatin‐like protein 2 in mouse brain mitochondria. European Journal Of Neuroscience 2013, 38: 2341-2348. PMID: 23617247, PMCID: PMC3902808, DOI: 10.1111/ejn.12237.Peer-Reviewed Original ResearchConceptsStomatin-like protein 2Type 1 receptorPresence of CB1Protein 2Anti-CB1 antibodySynthetic cannabinoid WINMouse brain mitochondriaCerebral cortexEndocannabinoid signalingBrain cellsCannabinoid WINNeuronal mitochondriaBrain mitochondriaAntibodiesMitochondrial functionCB1Polyclonal antibodiesCortexMitochondrial preparationsSerumReceptors
2011
Differential Acute and Chronic Effects of Leptin on Hypothalamic Astrocyte Morphology and Synaptic Protein Levels
García-Cáceres C, Fuente-Martín E, Burgos-Ramos E, Granado M, Frago LM, Barrios V, Horvath T, Argente J, Chowen JA. Differential Acute and Chronic Effects of Leptin on Hypothalamic Astrocyte Morphology and Synaptic Protein Levels. Endocrinology 2011, 152: 1809-1818. PMID: 21343257, PMCID: PMC3860256, DOI: 10.1210/en.2010-1252.Peer-Reviewed Original ResearchConceptsGlial fibrillary acidic proteinChronic leptin exposureSynaptic inputsAstrocyte morphologyLeptin exposureGFAP levelsGlial structural proteinsSynaptic protein densityChronic leptin administrationAcute leptin treatmentSynaptic protein levelsAdult male ratsCentral leptin resistanceFibrillary acidic proteinLevels 1 hPossible direct effectGlial ensheathingNeonatal overnutritionGlial activationLeptin levelsLeptin administrationHypothalamic neuronsLeptin resistanceLeptin treatmentMale ratsMaternal Ghrelin Deficiency Compromises Reproduction in Female Progeny through Altered Uterine Developmental Programming
Martin JR, Lieber SB, McGrath J, Shanabrough M, Horvath TL, Taylor HS. Maternal Ghrelin Deficiency Compromises Reproduction in Female Progeny through Altered Uterine Developmental Programming. Endocrinology 2011, 152: 2060-2066. PMID: 21325042, PMCID: PMC3075930, DOI: 10.1210/en.2010-1485.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsEmbryo ImplantationFemaleFertilityGene Expression Regulation, DevelopmentalGhrelinHeterozygoteHomeobox A10 ProteinsHomeodomain ProteinsImmunohistochemistryLitter SizeMaleMiceMice, KnockoutProliferating Cell Nuclear AntigenReproductionReverse Transcriptase Polymerase Chain ReactionTranscription FactorsUterusWnt ProteinsConceptsGhrelin deficiencyDevelopmental programmingAbnormal endometrial functionFemale wild-type miceUterus of miceLevels of ghrelinRegulation of appetiteWild-type miceReproductive tract developmentWild-type offspringSubsequent subfertilityEndometrial proliferationUnexposed miceEndometrial functionUtero exposureUterine expressionEmbryo implantationOvarian folliclesCorpora luteaGhrelinReproductive tractTract developmentMiceSignificant alterationsSubfertility
2010
Direct inhibition of hypocretin/orexin neurons in the lateral hypothalamus by nociceptin/orphanin FQ blocks stress-induced analgesia in rats
Gerashchenko D, Horvath TL, Xie X. Direct inhibition of hypocretin/orexin neurons in the lateral hypothalamus by nociceptin/orphanin FQ blocks stress-induced analgesia in rats. Neuropharmacology 2010, 60: 543-549. PMID: 21195099, PMCID: PMC3031765, DOI: 10.1016/j.neuropharm.2010.12.026.Peer-Reviewed Original ResearchMeSH KeywordsAnalgesiaAnalysis of VarianceAnimalsCell CountHypothalamic Area, LateralImmunohistochemistryIntracellular Signaling Peptides and ProteinsMaleMicroinjectionsMicroscopy, ElectronNeuronsNeuropeptidesOpioid PeptidesOrexinsPain MeasurementPain PerceptionProto-Oncogene Proteins c-fosRatsRats, Sprague-DawleyReceptors, OpioidRestraint, PhysicalStatistics, NonparametricStress, PhysiologicalConceptsStress-induced analgesiaHcrt neuronsLateral hypothalamusPerifornical areaNociceptin/orphanin FQ systemHypocretin/orexin neuronsNociceptin/orphanin FQHcrt neuronal activityDirect inhibitionThermal pain thresholdThermal nociceptive testsHypocretin/orexinFos immunohistochemistryOrexin neuronsBilateral microinjectionIntracerebroventricular injectionPain thresholdNociceptive testsOrphanin FQOFQ receptorMouse modelNeuronal activityBrain areasHypothalamusNeuronsSynaptic input organization of the melanocortin system predicts diet-induced hypothalamic reactive gliosis and obesity
Horvath TL, Sarman B, García-Cáceres C, Enriori PJ, Sotonyi P, Shanabrough M, Borok E, Argente J, Chowen JA, Perez-Tilve D, Pfluger PT, Brönneke HS, Levin BE, Diano S, Cowley MA, Tschöp MH. Synaptic input organization of the melanocortin system predicts diet-induced hypothalamic reactive gliosis and obesity. Proceedings Of The National Academy Of Sciences Of The United States Of America 2010, 107: 14875-14880. PMID: 20679202, PMCID: PMC2930476, DOI: 10.1073/pnas.1004282107.Peer-Reviewed Original ResearchConceptsHigh-fat dietSynaptic input organizationReactive gliosisPOMC neuronsDIO ratsDR ratsArcuate nucleusMelanocortin systemPOMC cellsNeuropeptide Y cellsInput organizationLoss of synapsesDiet-induced obesityBlood-brain barrierHFD-fed animalsDIO animalsAnorexigenic proopiomelanocortinGlial ensheathmentSynaptic organizationInhibitory inputsLean ratsDR animalsNeuronal circuitsCell bodiesGliosis
2008
Hypocretin/orexin and nociceptin/orphanin FQ coordinately regulate analgesia in a mouse model of stress-induced analgesia
Xie X, Wisor JP, Hara J, Crowder TL, LeWinter R, Khroyan TV, Yamanaka A, Diano S, Horvath TL, Sakurai T, Toll L, Kilduff TS. Hypocretin/orexin and nociceptin/orphanin FQ coordinately regulate analgesia in a mouse model of stress-induced analgesia. Journal Of Clinical Investigation 2008, 118: 2471-2481. PMID: 18551194, PMCID: PMC2423866, DOI: 10.1172/jci35115.Peer-Reviewed Original ResearchMeSH KeywordsAnalgesiaAnimalsAtaxin-3Behavior, AnimalBrainCalciumCytoplasmElectrophysiologyFemaleHypothalamus, PosteriorImmunohistochemistryIntracellular Signaling Peptides and ProteinsMaleMembrane PotentialsMiceMice, Inbred C57BLMice, TransgenicNarcotic AntagonistsNeuronsNeuropeptidesNociceptin ReceptorNuclear ProteinsOpioid PeptidesOrexinsPain ThresholdPresynaptic TerminalsReaction TimeReceptors, OpioidStress, PhysiologicalTetrodotoxinTranscription FactorsConceptsStress-induced analgesiaHcrt neuronsWild-type miceHypocretin/orexinNociceptin/orphanin FQMouse hypothalamic slicesCorticotropin-releasing factorPatch-clamp recordingsOrexin/ataxinPostsynaptic effectsPresynaptic releaseOrphanin FQElectron microscopic levelHypothalamic slicesSynaptic contactsHcrt-1Hcrt systemMouse modelAnalgesiaClamp recordingsPeptidergic systemsAction potentialsBrain tissueNeuronsInput resistance
2006
Uncoupling protein‐2 promotes nigrostriatal dopamine neuronal function
Andrews ZB, Rivera A, Elsworth JD, Roth RH, Agnati L, Gago B, Abizaid A, Schwartz M, Fuxe K, Horvath TL. Uncoupling protein‐2 promotes nigrostriatal dopamine neuronal function. European Journal Of Neuroscience 2006, 24: 32-36. PMID: 16882005, DOI: 10.1111/j.1460-9568.2006.04906.x.Peer-Reviewed Original ResearchMeSH Keywords3,4-Dihydroxyphenylacetic AcidAnimalsCorpus StriatumDopamineDopamine Plasma Membrane Transport ProteinsImmunohistochemistryIon ChannelsMaleMembrane Transport ProteinsMiceMice, KnockoutMitochondrial ProteinsMotor ActivityNeuronsSubstantia NigraTyrosine 3-MonooxygenaseUncoupling Protein 2ConceptsSubstantia nigra pars compactaDopamine neuronal functionUCP2-KO miceParkinson's diseaseNeuronal functionNigrostriatal dopamine functionTyrosine hydroxylase immunoreactivityUCP2 knockout miceDopamine transporter immunoreactivityProtein 2Wild-type controlsHydroxylase immunoreactivityPars compactaDopamine turnoverTransporter immunoreactivityDopamine ratioBehavioral deficitsLocomotor functionNucleus accumbensBiochemical deficitsDopamine functionBrain regionsNeurological pathologiesDiseaseMiceUncoupling protein 2/3 immunoreactivity and the ascending dopaminergic and noradrenergic neuronal systems: Relevance for volume transmission
Rivera A, Agnati LF, Horvath TL, Valderrama JJ, de La Calle A, Fuxe K. Uncoupling protein 2/3 immunoreactivity and the ascending dopaminergic and noradrenergic neuronal systems: Relevance for volume transmission. Neuroscience 2006, 137: 1447-1461. PMID: 16387447, DOI: 10.1016/j.neuroscience.2005.05.051.Peer-Reviewed Original ResearchConceptsConfocal laser microscopy analysisReactive oxygen species productionLaser microscopy analysisProtein 2/3Oxygen species productionUncouple oxidative phosphorylationOxidative phosphorylationATP synthesisProteinSpecies productionDouble immunolabelingImportant roleMicroscopy analysisPhosphorylationMitochondriaRegulationCell groupsPlastic changesLocalizationIslandsAnimal modelsMagnaTyrosine hydroxylaseNeuronal systems
2005
A Novel Growth Hormone Secretagogue-1a Receptor Antagonist That Blocks Ghrelin-Induced Growth Hormone Secretion but Induces Increased Body Weight Gain
Halem HA, Taylor JE, Dong JZ, Shen Y, Datta R, Abizaid A, Diano S, Horvath TL, Culler MD. A Novel Growth Hormone Secretagogue-1a Receptor Antagonist That Blocks Ghrelin-Induced Growth Hormone Secretion but Induces Increased Body Weight Gain. Neuroendocrinology 2005, 81: 339-349. PMID: 16210868, DOI: 10.1159/000088796.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsArea Under CurveBehavior, AnimalBinding, CompetitiveBody WeightBrainCell CountCHO CellsCricetinaeCricetulusDose-Response Relationship, DrugDrug InteractionsFeeding BehaviorGhrelinGrowth HormoneHumansImmunohistochemistryIodine IsotopesMaleOncogene Proteins v-fosPeptide HormonesRatsRats, Sprague-DawleyReceptors, G-Protein-CoupledReceptors, GhrelinTime FactorsConceptsDorsal medial hypothalamusGHS-1a receptorGrowth hormone secretionBIM-28163Fos-IRWeight gainGH secretionHormone secretionGrowth hormone secretagogue 1a receptorAntagonist of ghrelinMedial arcuate nucleusAction of ghrelinFos protein immunoreactivityAnti-obesity strategiesBody weight gainGhrelin activationConcomitant administrationGhrelin actionMedial hypothalamusArcuate nucleusReceptor antagonistGhrelin receptorFood intakeProtein immunoreactivityHuman ghrelinInput organization and plasticity of hypocretin neurons Possible clues to obesity’s association with insomnia
Horvath TL, Gao XB. Input organization and plasticity of hypocretin neurons Possible clues to obesity’s association with insomnia. Cell Metabolism 2005, 1: 279-286. PMID: 16054072, DOI: 10.1016/j.cmet.2005.03.003.Peer-Reviewed Original ResearchConceptsHypocretin neuronsSynaptic currentsHypothalamic hypocretin neuronsMore excitatory synapsesOvernight food deprivationElevated food intakeExcitatory synaptic currentsControl of arousalCause of narcolepsyStress-induced plasticityHypocretin levelsLeptin administrationHypocretin cellsHypocretin signalingMetabolic disturbancesObesity associationSynaptic organizationExcitatory synapsesFood intakeInhibitory inputsCell bodiesUnderlying causeNeuronsInput organizationFood deprivationUncoupling Protein-2 Is Critical for Nigral Dopamine Cell Survival in a Mouse Model of Parkinson's Disease
Andrews ZB, Horvath B, Barnstable CJ, Elseworth J, Yang L, Beal MF, Roth RH, Matthews RT, Horvath TL. Uncoupling Protein-2 Is Critical for Nigral Dopamine Cell Survival in a Mouse Model of Parkinson's Disease. Journal Of Neuroscience 2005, 25: 184-191. PMID: 15634780, PMCID: PMC6725213, DOI: 10.1523/jneurosci.4269-04.2005.Peer-Reviewed Original ResearchMeSH Keywords1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine1-Methyl-4-phenylpyridiniumAnimalsCell SurvivalCorpus StriatumDisease Models, AnimalDopamineHumansImmunohistochemistryIon ChannelsMaleMembrane Transport ProteinsMiceMice, Inbred C57BLMice, KnockoutMice, TransgenicMitochondriaMitochondrial ProteinsOxygen ConsumptionParkinsonian DisordersReactive Oxygen SpeciesSubstantia NigraUncoupling Protein 2ConceptsProtein 2Mitochondrial ROS productionLack of UCP2Reactive oxygen species productionGenetic manipulationOxygen species productionMitochondria numberCell metabolismATP synthesisCell survivalOverexpression of UCP2Wild-type controlsMitochondrial uncouplingNovel therapeutic targetROS productionUCP2Species productionElectron microscopic analysisOverexpressionCell functionUCP2 overexpressionDopamine cell survivalTherapeutic targetFluorescent ethidiumDopamine cell function
2004
Interaction between the Corticotropin-Releasing Factor System and Hypocretins (Orexins): A Novel Circuit Mediating Stress Response
Winsky-Sommerer R, Yamanaka A, Diano S, Borok E, Roberts AJ, Sakurai T, Kilduff TS, Horvath TL, de Lecea L. Interaction between the Corticotropin-Releasing Factor System and Hypocretins (Orexins): A Novel Circuit Mediating Stress Response. Journal Of Neuroscience 2004, 24: 11439-11448. PMID: 15601950, PMCID: PMC6730356, DOI: 10.1523/jneurosci.3459-04.2004.Peer-Reviewed Original ResearchMeSH KeywordsAction PotentialsAnimalsArousalBrainBrain ChemistryCorticotropin-Releasing HormoneFemaleHypothalamusImmunohistochemistryIn Vitro TechniquesIntracellular Signaling Peptides and ProteinsMaleMembrane PotentialsMiceMice, KnockoutNeural PathwaysNeuronsNeuropeptidesOrexin ReceptorsOrexinsReceptors, Corticotropin-Releasing HormoneReceptors, G-Protein-CoupledReceptors, NeuropeptideRecombinant Fusion ProteinsStress, PhysiologicalConceptsCorticotropin-releasing factorHypocretinergic neuronsHypocretin neuronsCorticotropin-Releasing Factor SystemCRF-immunoreactive terminalsHypocretin-expressing neuronsRelease of hypocretinsStability of arousalMaintenance of arousalHypocretinergic cellsHypothalamic slicesLateral hypothalamusAntagonist astressinHypocretinergic systemNeuropeptide hypocretinStressor stimuliPeptidergic systemsAcute stressHypocretinNeuronsStress responseFactor systemMembrane potentialPhysiological inputsActivationDirect visual and circadian pathways target neuroendocrine cells in primates
Abizaid A, Horvath B, Keefe DL, Leranth C, Horvath TL. Direct visual and circadian pathways target neuroendocrine cells in primates. European Journal Of Neuroscience 2004, 20: 2767-2776. PMID: 15548220, DOI: 10.1111/j.1460-9568.2004.03737.x.Peer-Reviewed Original ResearchConceptsSuprachiasmatic nucleusRetinal inputNeuroendocrine cellsDirect retinal inputHormone-releasing hormoneNon-human primatesHypothalamic suprachiasmatic nucleusSCN efferentsHypothalamic areaHypothalamic neuronsHypothalamic sitesMonosynaptic pathwayVisual afferentsHypothalamic regulatorGonadal axisHormone releaseNeuroendocrine functionPituitary gonadotropsPhotic modulationNeuronsCircadian pacemakerVervet monkeysPresent studyTracing techniquesCircadian clockSex differences in adult suprachiasmatic nucleus neurons emerging late prenatally in rats
Abizaid A, Mezei G, Sotonyi P, Horvath TL. Sex differences in adult suprachiasmatic nucleus neurons emerging late prenatally in rats. European Journal Of Neuroscience 2004, 19: 2488-2496. PMID: 15128402, DOI: 10.1111/j.0953-816x.2004.03359.x.Peer-Reviewed Original ResearchMeSH KeywordsAgingAnalysis of VarianceAnimalsAnimals, NewbornArginine VasopressinBromodeoxyuridineCalbindin 1CalbindinsCell CountEmbryo, MammalianFemaleGeniculate BodiesGlial Fibrillary Acidic ProteinImmunohistochemistryMaleNeuronsNeuropeptide YPregnancyPrenatal Exposure Delayed EffectsRatsRats, Sprague-DawleyS100 Calcium Binding Protein GSex CharacteristicsSuprachiasmatic NucleusTestosterone PropionateConceptsSuprachiasmatic nucleusGonadal steroidsFemale ratsLate gestationSex differencesPregnant female ratsVasoactive intestinal peptideGestational day 18Postnatal day 60BrdU-labeled cellsCalbindin-D28KSuprachiasmatic nucleus neuronsCircadian rhythmIntestinal peptideNucleus neuronsDouble immunocytochemistryGonadal functionTestosterone propionateBrain sectionsRat fetusesDay 18Day 60Cell groupsRatsSCN cellsPresynaptic N‐methyl‐D‐aspartate receptor expression is increased by estrogen in an aromatase‐rich area of the songbird hippocampus
Saldanha CJ, Schlinger BA, Micevych PE, Horvath TL. Presynaptic N‐methyl‐D‐aspartate receptor expression is increased by estrogen in an aromatase‐rich area of the songbird hippocampus. The Journal Of Comparative Neurology 2004, 469: 522-534. PMID: 14755533, DOI: 10.1002/cne.11035.Peer-Reviewed Original ResearchConceptsHP neuronsVertebrate hippocampusSongbird hippocampusN-methyl-d-aspartate receptor expressionSubcellular characteristicsInfluence of estrogenFunctional plasticityType glutamate receptorsZebra finch brainPrecise mechanismSomal sizeExcitatory pathwaysReceptor expressionExcitatory neurotransmissionExcitatory synapsesGlutamate receptorsPostsynaptic locusPresynaptic boutonsHippocampusEstrogenKey interfacePlasticityNeuronsAromataseAutoreception
2003
Estradiol affects axo-somatic contacts of neuroendocrine cells in the arcuate nucleus of adult rats
Parducz A, Zsarnovszky A, Naftolin F, Horvath TL. Estradiol affects axo-somatic contacts of neuroendocrine cells in the arcuate nucleus of adult rats. Neuroscience 2003, 117: 791-794. PMID: 12654332, DOI: 10.1016/s0306-4522(02)00967-3.Peer-Reviewed Original ResearchConceptsAxo-somatic contactsArcuate nucleusSynaptic plasticityGABAergic axo-somatic synapsesMorphological synaptic plasticityAxo-somatic synapsesEffects of estradiolSynapse quantificationArcuate neuronsTracer FluorogoldGonadal steroidsAnterior pituitaryAdult ratsHypophysiotropic neuronsMedian eminenceNervous systemTransient decreaseSystemic applicationNeuroendocrine cellsDisector methodNeuronsNumerical densityEstradiolFluorogoldCells