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首页> 外文期刊>Developmental biology >Qualitative and quantitative differences in protein synthesis comparing fetal lamb ductus arteriosus endothelium and smooth muscle with cells from adjacent vascular sites☆
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Qualitative and quantitative differences in protein synthesis comparing fetal lamb ductus arteriosus endothelium and smooth muscle with cells from adjacent vascular sites☆

机译:胎儿小动脉导管内皮细胞和平滑肌与相邻血管部位细胞相比,蛋白质合成的质和量差异☆

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Duringlategestation,intimalcushionsformintheductusarteriosus(DA)andthesecausethevesseltoclosewhenitconstrictsinthepostnatalperiod.TheformationofintimalcushionssuggestshighlyspecializedfunctionsofDAendothelialandsmoothmusclecells.Toinvestigatetheseproperties,weestablished,fromfetallambsonDay138ofa148-daytermgestation,primarycellculturesofDAendotheliumandsmoothmuscleandcomparedthemtocellsderivedfromtheadjacentpulmonaryarteryandaorta.Purityoftheendothelialcellculturesfromeachvascularsitewasassessedbythecontactinhibiteda€?cobblestonea€?monolayerphenotype,bypositiveimmunofluorescenceforfactorVIIIandbyangiotensinconvertingenzymeactivity.Purityofsmoothmusclecellculturesateachvascularsitewasassessedbythea€?hillsandvalleysa€?phenotypeandbypositiveimmunofluorescencewithasmoothmuscleactinspecificmonoclonalantibody.Endothelialandsmoothmusclecellshaddifferentgrowthcurves,ultrastructuralfeatures,andproteinprofilesonsingleandtwo-dimensionalSDS-polyacrylamidegelelectrophoresis(PAGE),butvascularsitesweresimilar.TofurtherdeterminewhetherdifferencesrelatedtoDAoriginwereindeedpresent,endothelialandsmoothmusclecellsfromallthreevascularsiteswereincubatedwiththeradiolabeledaminoacids[14C]leucine,[14C]proline,and[14C]valineandtheproteinsinboththecellsandtheconditionedmediumwereanalyzedbyautoradiographyafterSDS-PAGE.Adensebandcorrespondingtoa42-kDaproteinwasobservedinvaline-labeledDAendothelialcellsandconditionedmediumanda52-kDaproteinwasobservedintheconditionedmediumofleucine-labeledDAsmoothmusclecellsonly.FurtherisolationandcharacterizationoftheseendothelialandsmoothmuscleproteinswillbenecessarytodeterminewhethertheyarerelatedtothemechanismofintimalcushionformationinthelategestationDAorarepresentabnormallyinassociationwiththeintimalproliferationobservedinpulmonaryandsystemicvasculardisease.
机译:Duringlategestation,intimalcushionsformintheductusarteriosus(DA)andthesecausethevesseltoclosewhenitconstrictsinthepostnatalperiod.TheformationofintimalcushionssuggestshighlyspecializedfunctionsofDAendothelialandsmoothmusclecells.Toinvestigatetheseproperties,weestablished,fromfetallambsonDay138ofa148-daytermgestation,primarycellculturesofDAendotheliumandsmoothmuscleandcomparedthemtocellsderivedfromtheadjacentpulmonaryarteryandaorta.Purityoftheendothelialcellculturesfromeachvascularsitewasassessedbythecontactinhibiteda€?cobblestonea€?monolayerphenotype,bypositiveimmunofluorescenceforfactorVIIIandbyangiotensinconvertingenzymeactivity.Purityofsmoothmusclecellculturesateachvascularsitewasassessedbythea€?hillsandvalleysa€?phenotypeandbypositiveimmunofluorescencewithasmoothmuscleactinspecificmonoclonalantibody.Endothelialandsmoothmusclecellshaddifferentgrowthcurves,ultrastructuralfeatures,andproteinprofilesonsingleandtwo-dimensionalSDS-polyacrylamidegelelectrophoresis(PAGE) ,但血管部位resimilar.TofurtherdeterminewhetherdifferencesrelatedtoDAoriginwereindeedpresent,endothelialandsmoothmusclecellsfromallthreevascularsiteswereincubatedwiththeradiolabeledaminoacids [14 C]亮氨酸,[14 C]脯氨酸,和[14 C] valineandtheproteinsinboththecellsandtheconditionedmediumwereanalyzedbyautoradiographyafterSDS-PAGE.Adensebandcorrespondingtoa42-kDaproteinwasobservedinvaline-labeledDAendothelialcellsandconditionedmediumanda52-kDaproteinwasobservedintheconditionedmediumofleucine-labeledDAsmoothmusclecellsonly.FurtherisolationandcharacterizationoftheseendothelialandsmoothmuscleproteinswillbenecessarytodeterminewhethertheyarerelatedtothemechanismofintimalcushionformationinthelategestationDAorarepresentabnormallyinassociationwiththeintimalproliferationobservedinpulmonaryandsystemicvasculardisease。

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