Stimulation of NO production and of eNOS phosphorylation in the microcirculation <i>in vivo</i>

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dc.contributor.authorDurán, WN
dc.contributor.authorSeyama, A
dc.contributor.authorYoshimura, K
dc.contributor.authorGonzález, DR
dc.contributor.authorJara, PI
dc.contributor.authorFigueroa, XF
dc.contributor.authorBoric, MP
dc.date.accessioned2025-01-21T01:31:12Z
dc.date.available2025-01-21T01:31:12Z
dc.date.issued2000
dc.description.abstractThe role of nitric oxide (NO) in microvascular permeability is controversial, in part because the regulation of its endothelial constitutive synthase, eNOS, has been studied in vitro but not in vivo. Our study was designed to detect the morphologic and functional presence of eNOS and to test whether eNOS could be phosphorylated by platelet-activating factor (PAF), an agent that induces hyperpermeability. Immunocytochemistry was applied using human anti-eNOS antibodies in the hamster cheek pouch (hcp). The hcp microvessels demonstrated positive reaction products in the endothelium. The functional presence of eNOS in hcp was investigated by topical application of 10(-7) M PAF to the hcp and by measuring NO production by chemiluminescence. The mean baseline value of NO release was 63.3 +/- 6.9 pmol/ml (mean +/- SE), Application of PAF led to an increase in mean NO release to 120.8 +/- 31.2 pmol/ml (P < 0.05). In another series of experiments, 10(-7) M PAF was applied topically to hcp preincubated with [P-32]orthophosphoric acid. Immunoprecipitation and Western blots detected P-32-labeled bands that migrated with the mobility of positive eNOS indicating phosphorylated eNOS protein. The intensity of the radioactive bands was evaluated by computer;assisted image analysis. Comparison of the net band intensities yielded a mean PAF-treated/control ratio of 1.6 +/- 0.1. Our data demonstrate the morphologic and functional presence of eNOS in the microcirculation. The data also provide evidence that the function of microvascular eNOS is subject to regulation by phosphorylation. (C) 2000 Academic Press.
dc.description.funderNHLBI NIH HHS
dc.fuente.origenWOS
dc.identifier.doi10.1006/mvre.2000.2250
dc.identifier.issn0026-2862
dc.identifier.urihttps://doi.org/10.1006/mvre.2000.2250
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/97008
dc.identifier.wosidWOS:000089324400003
dc.issue.numero2
dc.language.isoen
dc.pagina.final111
dc.pagina.inicio104
dc.revistaMicrovascular research
dc.rightsacceso restringido
dc.subjecteNOS phosphorylation
dc.subjectmicrovasculature
dc.subjectmicrocirculation
dc.subjectWestern blot
dc.subjectimmunoprecipitation
dc.subjectplatelet-activating factor
dc.subjecthamster cheek pouch
dc.subjectNO determination
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleStimulation of NO production and of eNOS phosphorylation in the microcirculation <i>in vivo</i>
dc.typeartículo
dc.volumen60
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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